Abstract:
The present invention relates to a method for in vitro detection of SIRS, sepsis and/or sepsis-like conditions. This method renders the evaluation of the severity and/or the therapeutic progress of sepsis and severe infections, in particular sepsis-like systemic infections possible. Further, the present invention relates to the use of recombinantly or synthetically prepared nucleic acid sequences or peptide sequences derived therefrom as calibrator in sepsis assays and/or for the evaluation of the effect and the toxicity during screening of the active agents and/or the preparation of therapeutics for the prevention and treatment of SIRS, sepsis, sepsis-like systemic inflammatory conditions and sepsis-like systemic infections.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
       [0001]     This application is a National Stage of International Application PCT/EP04/03419, filed Mar. 31, 2004. International Application PCT/EP04/03419 cites for priority German application numbers 103 15 031.5 (filed Apr. 2, 2003), 103 36 511.7 (filed Aug. 8, 2003), and 103 40 395.7 (filed Sep. 2, 2003). This application incorporates by reference International Application PCT/EP04/03419, German application number 103 15 031.5, German Application Number 103 36 511.7, and German Application Number 103 40 395.7. This application incorporates by reference the Sequence Listing electronically submitted under file name “3535-027SuppSequence.TXT”, with the listed creation date of “May 7, 2007” and being “9,409 KB” in size. 
     
    
     BACKGROUND OF THE INVENTION  
       [0002]     The present invention relates to a method for in vitro detection of acute generalized inflammatory conditions (SIRS), sepsis, sepsis-like conditions, and systemic infections, as well as the use of recombinantly or synthetically prepared nucleic acid sequences or peptide sequences derived therefrom.  
         [0003]     Part of the description of the present invention is a sequence listing of 1430 pages, consisting of SEQ ID No: 1 through SEQ ID No: 10,540.  
         [0004]     The complete sequence listing is incorporated herein by reference, is part of the description and, thus, part of the disclosure of the present invention.  
         [0005]     The present invention particularly refers to labels for gene activity for the diagnosis and for the optimization of the therapy of acute generalized inflammatory conditions (Systemic Inflammatory Response Syndrome (SIRS)). Additionally, the present invention relates to methods for detecting acute generalized inflammatory conditions and/or sepsis, sepsis-like conditions, severe sepsis and systemic infections as well as for a corresponding improvement of therapy of acute generalized inflammatory conditions (SIRS).  
         [0006]     Further, for patients suffering from acute generalized inflammatory conditions (SIRS) the present invention relates to new possibilities of diagnosis that are obtained from experimentally proofed findings in connection with the occurrence of changes in gene activity (transcription and subsequent protein expression).  
         [0007]     In spite of the fact that there have been improvements of the pathophysiologic understanding and the supportive treatment of patients in intensive care units, SIRS is a disease that occurs very frequently and contributes considerably to mortality in patients in intensive care units [2-5].  
         [0008]     The criteria of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference (ACCP/SCCM) of 1992 are the ones that became most accepted in the international literature as definition of the term SIRS [4]. According to this definition, SIRS (in this patent described as acute generalized inflammatory conditions) is defined as systemic response of the inflammatory system triggered by a noninfectious stimulus. At least two of the following criteria have to be fulfilled in this context: Fever&gt;38° C. or hypothermia&lt;36° C., leukocytosis&gt;12 G/1 or leukopenia&lt;4 G/1 or shift to the left in the haemogram, heart rate&gt;90/min, tachypnoea&gt;20 breaths/min or PaCO2&lt;4.3 kPa, respectively.  
         [0009]     The mortality rate in SIRS amounts to about 20% and increases with the development of more severe organ dysfunctions [6]. The contribution of SIRS to morbidity and lethality is of multidisciplinary interest, as it increasingly puts the success of the most advanced or experimental treatment methods of many medicinal fields (e.g. cardiosurgery, traumatology, transplantation medicine, heamatology/onkology) at a risk, as they all are threatened by an increased risk of the development of an acute generalized inflammatory conditions. Thus, the decrease of morbidity and lethality of many seriously ill patients goes along with the improvement of prevention, treatment and particularly detection and observation of the progress of acute generalized inflammatory conditions.  
         [0010]     SIRS is a result of complex and very heterogeneous molecular processes that are characterized by the incorporation of many components and their interactions on every organizational level of the human body: genes, cells, tissues, organs. The complexity of the underlying biological and immunological processes resulted in many kinds of studies comprising a wide range of clinical aspects. One of the results from these studies was that the evaluation of new therapies is rendered more difficult due to the presently used criteria which are quite unspecific and clinical based and which do not sufficiently show the molecular mechanisms [7].  
         [0011]     Unfortunately, sepsis and consecutive organ dysfunctions still rank among the principal causes of death in non-cardiologic intensive care units [1-3]. It is supposed that 400,000 patients suffer from sepsis in the USA each year [4]. Lethality is about 40% and increases to 70-80% if a shock develops [5, 6]. The excess lethality independent from the underlying disease of the patient and the underlying infection amounts to 35% [8].  
         [0012]     The criteria of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference (ACCP/SCCM) of 1992 are the ones that became most accepted in the international literature as definition of the term sepsis [4]. According to these criteria [4] the grades of severity “systemic inflammatory response syndrom” (SIRS), “sepsis”, “severe sepsis” and “septic shock” are clinically defined. According to this definition, SIRS (in this patent described as acute generalized inflammatory conditions) is defined as the systemic response of the inflammatory system triggered by a noninfectious stimulus. At least two of the following criteria have to be fulfilled in this context: Fever&gt;38° C. or hypothermia&lt;36° C., leukocytosis&gt;12G/1 or leukopenia&lt;4G/1 or shift to the left in the haemogram, heart rate&gt;90/min, tachypnoea&gt;20 breaths/min or PaCO2&lt;4.3 kPa, respectively. According to the definition, sepsis are those clinical conditions in which the criteria of SIRS are fulfilled and an infection is detected as cause or it is at least very likely that it is the cause. A severe sepsis is characterized by the additional occurrence of organ dysfunctions. Frequent organ dysfunctions are changes in the state of consciousness, oliguria, lactate acidosis or sepsis-induced hypotension with a systolic blood pressure lower than 90 mmHg, or a pressure decrease of more than 40 mmHg of the initial value, respectively. If such a hypotension cannot be treated by administration of crystalloids and/or colloids and the patient further needs treatment with catecholamines, this is called a septic shock. Such a septic shock is detected in about 20% of all sepsis patients.  
         [0013]     Whether and how catecholamines are administered during the treatment of patients suffering from severe sepsis depends on the physician. If the blood pressure decreases, many physicians react by administering large quantities of infusion solutions and, thus, avoid administering catecholamines, however, there are also many physicians who refuse this kind of proceeding and who administer catecholamines much earlier and at a higher dose, if the patient shows the same clinical severity. The consequence is that in everyday practice patients suffering from the same clinical severity can be rated as belonging to the group “severe sepsis” or to the group “septic shock” [4] due to subjective reasons. This is why it became common in international literature to pool patients with the severity grades “severe sepsis” and “septic shock” [4] in one group. This is why the term “severe sepsis” used in this description is used according to the above mentioned consensus conference for patients with sepsis and additional proof of organ dysfunctions and, thus, comprises all patients of the groups “severe sepsis” and “septic shock” according to [4].  
         [0014]     The mortality rate in sepsis amounts to about 40% and increases to 70-80%, if a severe sepsis develops [5, 6]. The contribution of sepsis and severe sepsis to morbidity and lethality is of multidisciplinary interest. By comparison, the number of cases rose continuously (by 139% from 73.6 to 176 cases per 100,000 hospital patients from 1970 and 1977, for example) [7]. This increasingly puts the success of the most advanced or experimental treatment methods of many medicinal fields (e.g. visceral surgery, transplantation medicine, heamatology/onkology) at a risk, as they all are threatened by an increased risk of the development of acute generalized inflammatory conditions. Thus, the decrease of morbidity and lethality of many seriously ill patients goes along with a progress in prevention and treatment and especially detection and observation of the progress of the sepsis and severe sepsis. This is why well-known authors have been criticizing for a long time that too much energy and financial resources have been spend on the search for therapeutics for sepsis in the past decade, instead of using them for improving sepsis diagnosis.  
         [0015]     Sepsis is a result of complex and highly heterogeneous molecular processes that are characterized by the incorporation of many components and their interactions on every organizational level of the human body: genes, cells, tissues, organs. The complexity of the underlying biological and immunological processes resulted in many kinds of studies comprising a wide range of clinical aspects. One of the results from these studies was that the evaluation of new sepsis therapies is rendered more difficult due to the unspecific clinically based inclusioncriteria, which does not sufficiently show the molecular mechanisms [9].  
         [0016]     These facts have created need for innovative diagnostic means that are supposed to improve the capability of the person skilled in the art to diagnose patients suffering from SIRS, sepsis, sepsis-like conditions, severe sepsis and systemic infection at an early stage, to render the severity of a SIRS measurable on a molecular basis and to make it comparable in the clinical progress and to derive information concerning the individual prognosis and the reaction on specific treatments.  
         [0017]     The contribution of sepsis with regard to morbidity and lethality is of multidisciplinary interest. Lethality of sepsis changed only marginally within the last decades, whereas, in comparison, the indices increased continuously (e.g. from 1979 to 1987 by 139% from 73.6 to 176 per 100,000 in-patients) [7]. This increasingly puts the success of treatment of the most advanced or experimental therapy methods of various special fields (visceral surgery, transplantation medicine, heamatology/onkology) at a risk due to the fact that they all imply without exception an increase of the risk of sepsis. Thus, the decrease of morbidity and lethality of many seriously ill patients goes along with a progress in prevention and treatment and especially diagnosis of sepsis.  
         [0018]     Sepsis is a result highly heterogeneous molecular processes that are characterized by the incorporation of many components and their interactions on every organizational level of the human body: genes, cells, tissues, organs. The complexity of the underlying biological and immunological processes resulted in many kinds of studies comprising a wide range of clinical aspects. One of the results from these studies was that the evaluation of new sepsis therapies is rendered more difficult due to relatively unspecific clinically-based inclusioncriteria which do not sufficiently show the molecular mechanisms [9].  
         [0019]     Technological improvements, especially the development of microarray technology, are now rendering it possible for the person skilled in the art to compare 10 000 genes or more and their gene products at the same time. The use of such microarray technologies can now give hints on the conditions of health, regulation mechanisms, biochemical interactions and signalization networks. As the comprehension how an organism reacts to infections is improved this way, this should facilitate the development of enhanced modalities of detection, diagnosis and therapy of systemic disorders.  
         [0020]     Microarrays have their origin in “southern blotting” [10], the first approach to immobilize DNA-molecules so that it can be addressed three-dimensionally on a solid matrix. The first microarrays consisted of DNA-fragments, frequently with unknown sequence, and were applied dotwise onto a porous membrane (normally nylon). It was routine to use cDNA, genomic DNA or plasmid libraries, and to mark the hybridized material with a radioactive group [11-13].  
         [0021]     Recently, the use of glass as substrate and fluorescence for detection together with the development of new technologies for the synthesis and for the application of nucleic acids in very high densities allowed the miniaturizing of the nucleic acid arrays. At the same time, the experimental throughput and the information content were increased [14-16].  
         [0022]     Further, it is known from WO 03/002763 that microarrays basically can be used for the diagnosis of sepsis and sepsis-like conditions.  
         [0023]     The first explanation for the applicability of microarray technology was obtained through clinical studies on the field of cancer research. Here, expression profiles proofed to be valuable with regard to identification of activities of individual genes or groups of genes, correlating with certain clinical phenotypes [17]. Many samples of individuals with or without leukemia or diffuse lymphoma of large B-cells were analyzed and gene expression labels (RNA) were found and used for the classification of those kinds of cancer [17, 18]. Golub et al. found out that an individual gene is not enough to make reliable predictions, however, that predictions made on gene expression profiles of 53 genes (selected from more than 6000 genes that were present on the arrays) are highly accurate [17].  
         [0024]     Alisadeh et al. [18] examined large B-cell lymphoma (DLBCL). The authors created expression profiles with a “lymph chip”, a microarray bearing 18 000 clones of complementary DNA that was developed to monitor genes that are involved in normal and abnormal development of lymphocytes. By using cluster analysis, they managed to classify DILBCL in two categories that showed great differences with regard to the survival chance of patients. The gene expression profiles of these subtypes corresponded to two important stages of differentiation of B-cells.  
         [0025]     To differentiate between symptoms that base on microbial infections and other symptoms of non-infectious etiology, which could indicate sepsis due to their clinical appearance, but are in fact not based on a systemic microbial infection, e.g. of symptoms that base on non-infectious inflammation of individual organs, the determination of gene expression profiles via differential diagnostics proofed to be particularly advantageous [19-22]. The use of body fluids for the measurement of gene expression profiles for the diagnosis of SIRS has not yet been described.  
         [0026]     The point of origin of the invention disclosed in the present patent application is the realization that RNA levels different from normal values respectively peptide levels or peptide segment levels derivable from the RNA levels, that can be detected in a serum or plasma of a patient whose risk is high that he will be suffering from SIRS, or who suffers from symptoms that are typical for SIRS, can be detected before SIRS, sepsis, sepsis-like conditions, severe sepsis and systemic Infections are detected in biological samples.  
         [0027]     Thus, it is an object of the present invention to provide a method for the detection, evaluation of the degree of severity, and/or the progress of the therapy, of SIRS and/or sepsis and/or severe sepsis and/or systemic infections.  
         [0028]     The method of the invention is characterized in that the activity of one or more leading genes can be determined in a sample of a biological liquid of an individual. Additionally, SIRS and/or the success of a therapeutic treatment can be deduced from the presence and/or, if present, the amount of the determined gene product.  
         [0029]     One embodiment of the present invention is characterized in that the method for in vitro detection of SIRS comprises the following steps: 
    a) Isolation of sample RNA from a sample of a mammal;     b) Labelling of the sample RNA and/or at least one DNA being a gene or gene fragment specific for SIRS, with a detectable label.     c) Contacting the sample RNA with the DNA under hybridization conditions;     d) Contacting control RNA representing a control for non-pathologic conditions, with at least one DNA, under hybridization conditions, whereby the DNA is a gene or gene fragment specific for SIRS;     e) Quantitative detection of the label signals of the hybridized sample RNA and control RNA;     f) Comparing the quantitative data of the label signals in order to determine whether the genes or gene fragments specific for SIRS are more expressed in the sample than in the control, or less.    
 
         [0036]     One alternative embodiment of the present invention is characterized in that the method for in vitro detection of sepsis and/or sepsis-like conditions comprises the following steps: 
    g) Isolation of sample RNA from a sample of a mammal;     h) Labelling of the sample RNA and/or at least one DNA being a specific gene or gene fragment for sepsis and/or sepsis-like conditions, with a detectable label.     i) Contacting the sample RNA with the DNA under hybridization conditions;     j) Contacting sample RNA representing a control for non-pathologic conditions, with at least one DNA, under hybridization conditions, whereby the DNA is a gene or gene fragment specific for sepsis and/or sepsis-like conditions;     k) Quantitative detection of the label signals of the hybridized sample RNA and control RNA;     l) Comparing the quantitative data of the marking signals in order to determine whether the genes or gene fragments specific for sepsis and/or sepsis-like conditions are more expressed in the sample than in the control, or less.    
 
         [0043]     One embodiment of the present invention is characterized in that the method for in vitro detection of severe sepsis comprises the following steps: 
    m) Isolation of sample RNA from a sample of a mammal;     n) Labelling of the sample RNA and/or at least one DNA being a specific gene or gene fragment for severe sepsis, with a detectable label.     o) Contacting the sample RNA with the DNA under hybridization conditions;     p) Contacting sample RNA representing a control for non-pathologic conditions, with at least one DNA, under hybridization conditions, whereby the DNA is a gene or gene fragment specific for severe sepsis;     q) Quantitative detection of the label signals of the hybridized sample RNA and control RNA;     r) Comparing the quantitative data of the label signals in order to determine whether the genes or gene fragments specific for severe sepsis are more expressed in the sample than in the control, or less.    
 
         [0050]     A further embodiment of the present invention is characterized in that the control RNA is hybridized with the DNA before the measurement of the sample RNA and the label signals of the control RNA/DNA complex is gathered and, if necessary, recorded in form of a calibration curve or table.  
         [0051]     Another embodiment of the present invention is characterized in that mRNA is used as sample RNA.  
         [0052]     Another embodiment of the present invention is characterized in that the DNA is arranged, particularly immobilized, on predetermined areas on a carrier in form of a microarray.  
         [0053]     Another embodiment of the invention is characterized in that the method is used for early detection by means of differential diagnostics, for control of the therapeutic progress, for risk evaluation for patients as well as for post mortem diagnosis of SIRS and/or sepsis and/or severe sepsis and/or systemic infections and/or septic conditions and/or infections.  
         [0054]     Another embodiment of the present invention is characterized in that the sample is selected from: body fluids, in particular blood, liquor, urine, ascitic fluid, seminal fluid, saliva, puncture fluid, cell content, or a mixture thereof.  
         [0055]     Another embodiment of the present invention is characterized in that cell samples are subjected a lytic treatment, if necessary, in order to free their cell contents.  
         [0056]     Another embodiment of the present invention is characterized in that the mammal is a human.  
         [0057]     Another embodiment of the invention is characterized in that the gene or gene segment specific for SIRS is selected from the group consisting of SEQ. ID No. 6373 to SEQ. ID No. 10540, as well as from gene fragments thereof having at least 5-2000, preferably 20-200, more preferably 20-80 nucleotides.  
         [0058]     Another embodiment of the invention is characterized in that the gene or gene segment specific for sepsis and/or sepsis-like conditions is selected from the group consisting of SEQ. ID No. 1 to SEQ. ID No. 6242, as well as gene fragments thereof with 5-2000 or more, preferably 20-200, more preferably 20-80 nucleotides.  
         [0059]     Another embodiment of the invention is characterized in that the gene or gene segment specific for severe sepsis is selected from the group consisting of SEQ. ID No. 6243 to SEQ. ID No. 6372, as well as gene fragments thereof with 5-2000 or more, preferably 20-200, more preferably 20-80 nucleotides.  
         [0060]     Another embodiment of the present invention is characterized in that the immobilized probes are labelled. As probes for this embodiment serve self-complementary oligonucleotides, so called molecular beacons. They bear a fluorophore/quencher pair at their ends, so that they are present in a folded hairpin structure and only deliver a fluorescence signal with corresponding sample sequence. The hairpin structure of the molecular beacons is stable until the sample hybridizes at the specific catcher sequence, this leading to a change in conformation and, thus, to the release of reporter fluorescence.  
         [0061]     Another embodiment of the present invention is characterized in that at least 2 to 100 different cDNAs are used.  
         [0062]     Another embodiment of the present invention is characterized in that at least 200 different cDNAs are used.  
         [0063]     Another embodiment of the present invention is characterized in that at least 200 to 500 different cDNAs are used.  
         [0064]     Another embodiment of the present invention is characterized in that at least 500 to 1000 different cDNAs are used.  
         [0065]     Another embodiment of the present invention is characterized in that at least 1000 to 2000 different cDNAs are used.  
         [0066]     Another embodiment of the present invention is characterized in that the cDNA of the genes listed in claim  10  is replaced by synthetic analoga as well as peptidonucleic acids.  
         [0067]     Another embodiment of the present invention is characterized in that the synthetic analoga of the genes comprise 5-100, in particular about 70 base pairs.  
         [0068]     Another embodiment of the present invention is characterized in that a radioactive label is used as detectable label, in particular  32 P,  14 C,  125 I,  155 Eu,  33 P or  3 H.  
         [0069]     Another embodiment of the present invention is characterized in that a non-radioactive label is used as detectable label, in particular a color- or fluorescence label, an enzyme label or immune label, and/or quantum dots or an electrically measurable signal, in particular the change in potential, and/or conductivity and/or capacity during hybridizations.  
         [0070]     Another embodiment of the present invention is characterized in that the sample RNA and control RNA bear the same label.  
         [0071]     Another embodiment of the present invention is characterized in that the sample RNA and control RNA bear different labels.  
         [0072]     Another embodiment of the present invention is characterized in that the cDNA probes are immobilized on glass or plastics.  
         [0073]     Another embodiment of the present invention is characterized in that the individual cDNA molecules are immobilized onto the carrier material by means of a covalent binding.  
         [0074]     Another embodiment of the present invention is characterized in that the individual cDNA molecules are immobilized onto the carrier material by means of adsorption, in particular by means of electrostatic and/or dipole-dipole and/or hydrophobic interactions and/or hydrogen bridges.  
         [0075]     Another embodiment of the method according to the present invention for in vitro detection of SIRS is characterized in that it comprises the following steps: 
    a) Isolation of sample peptides from a sample of a mammal;     b) Labelling of the sample peptides with a detectable label;     c) Contacting the labelled sample peptides with at least one antibody or its binding fragment, whereby the antibody binds a peptide or peptide fragment specific for SIRS;     d) Contacting the labelled control peptides originating from healthy subjects, with at least one antibody or its binding fragment immobilized in form of a microarray on a carrier, whereby the antibody binds a peptide or peptide fragment specific for SIRS;     e) Quantitative detection of the label signals of the sample peptides and the control peptides;     f) Comparing the quantitative data of the label signals in order to determine whether the genes or gene fragments specific for SIRS are more expressed in the sample than in the control, or less.    
 
         [0082]     Another alternative embodiment of the method according to the present invention for in vitro detection of sepsis and/or sepsis-like conditions is characterized in comprising the following steps: 
        g) Isolation of sample peptides from a sample of a mammal;     h) Labelling of the sample peptides with a detectable label;     i) Contacting the labelled sample peptides with at least one antibody or its binding fragment, whereby the antibody binds a peptide or peptide fragment specific for sepsis and/or sepsis-like conditions;     j) Contacting the labelled control peptides originating from healthy subjects, with at least one antibody or its binding fragment immobilized on a carrier in form of a microarray, whereby the antibody binds a peptide or peptide fragment specific for sepsis and/or sepsis-like conditions;     k) Quantitative detection of the label signals of the sample peptides and the control peptides;     l) Comparing the quantitative data of the label signals in order to determine whether the genes or gene fragments specific for sepsis and/or sepsis-like conditions are more expressed in the sample than in the control, or less.        
 
         [0089]     Another embodiment of the method according to the present invention for in vitro detection of severe sepsis is characterized in comprising the following steps: 
    m) Isolation of sample peptides from a sample of a mammal;     n) Labelling of the sample peptides with a detectable label;     o) Contacting the labelled sample peptides with at least one antibody or its binding fragment, whereby the antibody binds a peptide or peptide fragment specific for severe sepsis;     p) Contacting the labelled control peptides stemming from healthy subjects, with at least one antibody or its binding fragment immobilized on a carrier in form of a microarray, whereby the antibody binds a peptide or peptide fragment specific for severe sepsis;     q) Quantitative detection of the label signals of the sample peptides and the control peptides;     r) Comparing the quantitative data of the label signals in order to determine whether the genes or gene fragments specific for severe sepsis are more expressed in the sample than in the control, or less.    
 
         [0096]     Another embodiment of the present invention is characterized in that the antibody is immobilized on a carrier in form of a microarray.  
         [0097]     Another embodiment of the present invention is characterized in providing an immunoassay.  
         [0098]     Another embodiment of the invention is characterized in that the method is used for early detection by means of differential diagnostics, for control of the therapeutic progress, for risk evaluation for patients as well as for post mortem diagnosis of SIRS and/or sepsis and/or severe sepsis and/or systemic infections.  
         [0099]     Another embodiment of the present invention is characterized in that the sample is selected from: body fluids, in particular blood, liquor, urine, ascitic fluid, seminal fluid, saliva, puncture fluid, cell content, or a mixture thereof.  
         [0100]     Another embodiment of the present invention is characterized in that tissue- and cell samples are subjected to a lytic treatment, if necessary, in order to free the content of the cells.  
         [0101]     Another embodiment of the present invention is characterized in that the mammal is a human.  
         [0102]     Another embodiment of the invention is characterized in that the peptide specific for SIRS is an expression product of a gene or gene fragment selected from the group consisting of SEQ. ID No. 6373 to SEQ. ID No. 10540, as well as gene fragments thereof with 5-2000 or more, preferably 20-200, more preferably 20-80 nucleotides.  
         [0103]     Another embodiment of the invention is characterized in that the peptide specific for sepsis and/or sepsis-like conditions is an expression product of a gene or gene fragment selected from the group consisting of SEQ. ID No. 1 to SEQ. ID No. 6242, as well as gene fragments thereof with 5-2000 nucleotides or more, preferably 20-200, more preferable 20-80 nucleotides.  
         [0104]     Another embodiment of the invention is characterized in that the peptide specific for severe sepsis is an expression product of a gene or gene fragment selected from the group consisting of SEQ. ID No. 6243 to SEQ. ID No. 6372, as well as gene fragments thereof with 5-2000 or more, preferably 20-200, more preferably 20-80 nucleotides.  
         [0105]     Another embodiment of the present invention is characterized in that at least 2 to 100 different peptides are used.  
         [0106]     Another embodiment of the present invention is characterized in that at least 200 different peptides are used.  
         [0107]     Another embodiment of the present invention is characterized in that at least 200 to 500 different peptides are used.  
         [0108]     Another embodiment of the present invention is characterized in that at least 500 to 1000 different peptides are used.  
         [0109]     Another embodiment of the present invention is characterized in that at least 1000 to 2000 different peptides are used.  
         [0110]     Another embodiment of the present invention is characterized in that a radioactive label is used as detectable label, in particular  32 P,  14 C,  125 I,  155 Eu,  33 P or  3 H.  
         [0111]     Another embodiment of the present invention is characterized in that a non-radioactive label is used as detectable label, in particular a color- or fluorescence label, an enzyme label or immune label, and/or quantum dots or an electrically measurable signal, in particular the change in potential, and/or conductivity and/or capacity during hybridizations.  
         [0112]     Another embodiment of the present invention is characterized in that the sample peptides and control peptides bear the same label.  
         [0113]     Another embodiment of the present invention is characterized in that the sample peptides and control peptides bear different labels.  
         [0114]     Another embodiment of the present invention is characterized in that the peptide probes are immobilized on glass or plastics.  
         [0115]     Another embodiment of the present invention is characterized in that the individual peptide molecules are immobilized onto the carrier material by means of a covalent binding.  
         [0116]     Another embodiment of the present invention is characterized in that the individual peptide molecules are immobilized on the carrier material by means of adsorption, in particular by means of electrostatic and/or dipole-dipole and/or hydrophobic interactions and/or hydrogen bridges.  
         [0117]     Another embodiment of the present invention is characterized in that the individual peptide molecules are detected by means of monoclonal antibodies or their binding fragments.  
         [0118]     Another embodiment of the present invention is characterized in that the determination of individual peptides by means of immunoassay or precipitation assay is carried out using monoclonal antibodies.  
         [0119]     Another embodiment of the present invention is the use of recombinantly or synthetically produced nucleic acid sequences, partial sequences or protein-/peptide-sequences derived thereof, specific for SIRS, individually or as partial quantities as calibrator in SIRS-assays and/or to evaluate the effects and toxicity when screening for active agents and/or for the preparation of therapeutics as well as of substances and compounds that are designed to act as therapeutics, for prevention and treatment of SIRS.  
         [0120]     Another embodiment of the present invention is the use of recombinantly or synthetically produced nucleic acid sequences, partial sequences or protein-/peptide-sequences derived thereof, specific for sepsis and/or sepsis-like conditions, individually or as partial quantities as calibrator in sepsis-assays and/or to evaluate the effects and toxicity when screening for active agents and/or for the preparation of therapeutics as well as of substances and compounds that are designed to act as therapeutics, for prevention and treatment of sepsis, sepsis-like systemic inflammatory conditions and sepsis-like systemic infections.  
         [0121]     Another embodiment of the present invention is the use of recombinantly or synthetically produced nucleic acid sequences, partial sequences or protein-/peptide-sequences derived thereof, specific for severe sepsis, individually or as partial quantities as calibrator in sepsis-assays and/or to evaluate the effects and toxicity when screening for active agents and/or for the preparation of therapeutics as well as of substances and compounds that are designed to act as therapeutics, for prevention and treatment of severe sepsis.  
         [0122]     It is obvious to the person skilled in the art that the individual features of the present invention shown in the claims can be combined with each other in any desired way.  
         [0123]     The term leading genes as used in the present invention means all derived DNA-sequences, partial sequences and synthetic analoga (for example peptido-nucleic acids, PNA). In the present invention, it further means all proteins, peptides or partial sequences, respectively, or synthetic peptide mimetics decoded by leading genes are meant. The description of the invention referring to the determination of the gene expression is not a restriction but only an exemplary application of the present invention.  
         [0124]     The description of the invention referring to blood is only an exemplary embodiment of the present invention. The term biological liquids as used in the present invention means all human body fluids.  
         [0125]     One application of the method according to the invention is the measurement of differential gene expression with SIRS, sepsis, sepsis-like conditions, severe sepsis and systemic infections. For this measurement, the RNA is isolated from the whole blood of corresponding patients and a control sample of a healthy subject or of a subject that is not suffering from one of the above-mentioned disorders. Subsequently, the RNA is labelled, for example radioactively with  32 P or with dye molecules (fluorescence). All molecules and/or detection signals known in the state of the art for labelling molecules may be used as labelling molecules. The person skilled in the art is also aware of the corresponding molecules and/or methods.  
         [0126]     The RNA thus labelled is subsequently hybridized with cDNA-molecules that are immobilized on a microarray. The cDNA-molecules immobilized on the microarray are a specific selection of genes according to claim  12  of the present invention for the measurement of SIRS, according to claim  13  for sepsis and sepsis-like conditions, according to claim  14  for severe sepsis and systemic infections.  
         [0127]     The intensity signals of the hybridized molecules are measured afterwards by means of suitable instruments (phosphorimager, microarray scanner) and analyzed by means of additional computer-based analysis. The expression ratios of the sample of the patient and the control are determined with the signal intensities measured. The expression ratios of the under- and/or overregulated genes indicate, as in the experiments described below, whether SIRS, sepsis, sepsis-like conditions, severe sepsis and systemic infections are present or not.  
         [0128]     Another use of the method according to the invention is the measurement of the differential gene expression to determine how probable it is that the patient will respond to the planned therapy, and/or for determination of the reaction to a specialized therapy and/or the settlement of the end of the therapy in terms of a “drug monitoring” with patients suffering from SIRS, sepsis, sepsis-like conditions, severe sepsis and systemic infections. For this purpose, the RNA (sample RNA) is isolated from the blood samples of the patient, that have been taken in time intervals. The different RNA samples are labelled together with the control sample and hybridized with the selected genes that are immobilized on a microarray. Thus, the corresponding expression ratios show the probability that patients respond to the planned therapy, and/or whether the started therapy is effective, and/or how long the patients&#39; treatment has to go on, and/or whether the maximum effect of the therapy has already been achieved with the dose and duration applied.  
         [0129]     Another use of the method according to the invention is the measurement of the binding grade of proteins, for example monoclonal antibodies, by means of the use of immunoassays, protein- and peptide arrays or precipitation assays. Durch die Bestimmung der Konzentration der von den Sequenzen der in Anwendungsbeispiel 1 aufgeführten Nukleinsäuren entsprechenden Proteine or Peptide kann auf ein erhöhtes Risiko zur Entwicklung einer SIRS geschlossen werden. Additionally, this procedure allows the differential diagnostic determination in patients suffering from SIRS, sepsis, sepsis-like conditions, severe sepsis and systemic infections.  
         [0130]     Additionally, this indicates a higher risk of development of sepsis, sepsis-like conditions, severe sepsis and systemic infections.  
         [0131]     Further advantages and features of the present invention will become apparent from the description of the embodiments as well as from the drawing. 
     
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0132]      FIG. 1  is a 2-dimensional gel comprising a precipitated serum protein of a patient suffering from sepsis that is applied to it.  
         [0133]      FIG. 2  is a 2-dimensional gel comprising a precipitated serum protein of a control that is applied to it. 
     
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
     Embodiment 1  
     SIRS  
       [0134]     Studies of differential gene expression with patients suffering from SIRS.  
         [0135]     Whole blood samples of patients who were under the care of a surgical intensive care unit were examined for the measurement of the differential gene expression in connection with SIRS.  
         [0136]     Control samples were whole blood samples of the patients that were drawn immediately before the operation. No one of these patients showed an infection and/or clinical signs of SIRS (defined according to the SIRS-criteria [4]) at this point of time or before the stationary treatment.  
         [0137]     Additionally, whole blood samples of the same patients who had been subjected to a surgery, were drawn four hours after the operation (patient samples). Each of these patients developed SIRS after the surgery. A range of characteristics of the patients suffering from SIRS are shown in table 1. In Table 1, data with regard to age, gender, diagnosis as well as duration of the extracorporeal treatment are given.  
                                     TABLE 1                           Data of the group of patients                            Duration of                       extracorporeal       Patient   Gender   Age   Diagnosis   treatment [min]               1   male   57   coronary heart disease   82       2   male   70   coronary heart disease   83       3   female   67   coronary heart disease   72       4   male   70   coronary heart disease   55                  
 
         [0138]     After the whole blood had been drawn, the total RNA was isolated using the PAXGene Blood RNA Kit according to the producer&#39;s (Quiagen) instructions. Subsequently, the cDNA was synthesized from the total RNA by means of reverse transcriptions with Superscript II RT (Invitrogen) according to the producer&#39;s instructions, labelled with aminoallyl-dUTP and succinimidylester of the fluorescent dyes Cy3 and Cy5 (Amersham), and hydrolyzed.  
         [0139]     The microarrays (Lab-Arraytor human 500-1 cDNA) of the company SIRS-Lab GmbH were used for the hybridization. These micorarrays are loaded with 340 humane cDNA-molecules. The 340 humane cDNA-molecules are 3-fold immobilized in three subarrays on each microarray.  
         [0140]     The prepared and labelled samples were hybridized with the microarrays according to the producer&#39;s instructions and subsequently washed. The fluorescence signals of the hybridized molecules were measured by means of a scanner (AXON 4000B).  
         [0000]     Analysis  
         [0141]     One test was analyzed by means of scanned pictures of the microarrays after hybridization. The mean intensity value of the detected spots was defined as the measured expression value of the corresponding gene. Spots were automatically identified and their homogeneity was checked. The analysis was controlled manually. In addition to the desired information, namely the amount of nucleic acids bound, contain the detected signals also background signals which are caused by unspecific bindings to the surface of the membrane. The definition of the signals of the background rendered the optimum differentiation between spots and the surface of the chip possible, which also showed color effects. For the analysis of the microarrays blank spots were chosen as background. The mean expression value of the chosen blank spots within one block (of 14 times 14 spots) was subtracted from the expression values of the gene spots (in the corresponding block).  
         [0142]     Point signals not caused by binding of nucleic acids but by dust particles or other disturbances on the filter, could be told from real spots because of their irregular shape and were excluded from further analysis.  
         [0143]     In order to render the values between the 3 subarrays and between different microarrays comparable, it was necessary to normalize the data afterwards. Due to the high number of spots on the microarray, the mean value of all expression values was set as normalization reference. The mean expression per gene was calculated by choosing the two (from three) repetitions that were closest to each other.  
         [0144]     The expression ratios of the samples of the control and the patients were calculated from the signal intensities using the software AIDA Array Evaluation. The criteria for the grading of the examined genes was the level of the expression ratio. The interesting genes were those which were most overexpressed or underexpressed, respectively, compared with the control samples.  
         [0145]     Table 2 shows that 57 genes of the patient sample were found, which were significantly overexpressed, if compared with the control sample. Table 3 shows that 16 genes of the patient sample were found, which were significantly underexpressed, if compared with the control sample. Those results show that the genes listed in table 2 and table 3 correlate with the occurrence of SIRS. Thus, the gene activities of the genes mentioned are labels for a diagnosis of SIRS.  
                                                                           TABLE 2                           Significantly increased transcription activities and       their relative ratio to the control sample in SIRS            GenBank                       SEQUENCE-       Accession-No.   Hugo-Name   Patient 1   Patient 2   Patient 3   Patient 4   ID                    XM_051958   ALOX5   2.43   1.49   1.81   1.40   6408       XM_015396   ALOX5AP   3.71   7.39   3.89   2.68   6409       XM_008738   BCL2   1.16   6.76   1.55   1.04   6410       BC016281   BCL2A1   13.71   10.29   1.41   4.36   6468       NM_021073   BMP5   2.02   1.83   1.78   1.51   6411       XM_002101   BMP8   2.32   10.85   1.31   0.87   6412       XM_045933   CAMKK2   2.20   1.26   1.95   1.13   6413       XM_055386   CASP1   1.40   1.76   1.89   1.45   6414       NM_004347   CASP5   1.92   2.77   0.67   1.89   6415       NM_004166   CCL14   1.24   1.58   2.46   0.77   6463       XM_012649   SCYA7   1.24   9.78   0.85   1.82   6465       NM_001760   CCND3   1.23   2.68   1.56   1.12   6416       NM_000591   CD14   3.45   4.43   1.76   2.05   6417       XM_038773   CD164   0.84   1.91   3.26   3.15   6418       XM_048792   CD1A   3.24   3.10   1.00   1.11   6419       NM_001779   CD58   2.14   2.11   1.54   2.91   6420       XM_002948   CD80   1.69   1.16   2.25   0.69   6423       XM_027978   CFLAR   2.33   4.97   1.44   1.39   6424       NM_000760   CSF3R   1.55   1.47   1.81   1.02   6425       XM_012717   CSNK1D   1.95   3.15   1.24   1.32   6426       XM_048068   SCYD1   3.70   12.12   0.86   3.88   6466       XM_051229   CXCR4   2.33   2.10   2.15   1.60   6427       XM_039625   DUSP10   2.49   3.77   0.90   1.10   6429       XM_010177   DUSP9   2.17   5.27   1.12   1.63   6430       XM_055699   ENTPD1   1.91   3.18   0.71   0.86   6431       XM_007189   FOXO1A   1.61   3.10   1.09   1.67   6432       XM_012039   FUT4   1.55   5.07   1.88   0.93   6433       XM_040683   HPRT1   5.15   66.19   1.44   2.28   6434       NM_017526   OBRGRP   1.93   1.10   1.53   1.40   6435       XM_049516   ICAM1   1.27   1.88   2.05   1.30   6436       XM_049531   ICAM3   2.31   2.32   1.61   1.45   6437       XM_041744   IER3   4.17   7.25   1.98   2.08   6438       XM_048562   IFNAR1   2.16   4.87   1.09   2.36   6439       XM_006447   IL10RA   1.02   1.51   1.96   0.67   6440       M90391   IL-16   1.77   1.50   1.16   1.09   6441       XM_002765   IL1R2   2.84   12.75   1.03   2.75   6442       NM_000418   IL4R   3.34   6.44   2.05   2.79   6443       XM_057491   IL6   1.72   1.48   1.53   1.37   6444       NM_002184   IL6ST   2.50   9.25   1.07   1.87   6445       NM_000634   IL8RA   2.27   3.73   1.45   1.68   6446       NM_006084   ISGF3G   1.72   1.08   2.54   1.12   6447       XM_045985   ITGA2B   3.69   2.00   0.83   3.79   6448       XM_008432   ITGA3   2.11   7.62   1.08   1.06   6449       XM_028642   ITGA5   2.49   4.48   1.39   3.54   6450       XM_036107   ITGB2   1.72   1.13   2.08   1.13   6451       XM_009064   JUNB   2.21   1.84   3.59   2.05   6452       XM_036154   LAMP2   1.79   1.68   1.62   1.41   6453       XM_042066   MAP3K1   2.06   7.67   2.91   8.93   6454       NM_001315   MAPK14   2.50   12.01   0.90   4.20   6455       NM_003684   MKNK1   2.58   17.17   1.74   1.83   6456       U68162   MPL   2.58   1.10   1.39   6.99   6457       NM_004555   NFATC3   1.40   1.70   2.80   0.75   6458       XM_006931   OLR1   1.53   5.01   1.10   3.16   6459       XM_039764   PDCD5   1.11   3.09   1.21   1.95   6460       XM_029791   PIK3C2G   0.93   1.62   0.96   1.52   6461       NM_006219   PIK3CB   1.52   0.99   0.94   1.66   6467       XM_043864   PIK3R1   1.81   4.07   1.48   1.26   6462                  
 
         [0146]    
       
         
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                   
               
               
                 Significantly reduced transcription activities and 
               
               
                 their relative ratio to the control sample in SIRS 
               
             
          
           
               
                 GenBank 
                   
                   
                   
                   
                   
                 SEQUENCE- 
               
               
                 Accession-No. 
                 HUGO Name 
                 Patient 1: 
                 Patient 2: 
                 Patient 3: 
                 Patient 4: 
                 ID 
               
               
                   
               
               
                 BC001374 
                 CD151 
                 0.00 
                 0.00 
                 0.39 
                 0.71 
                 6375 
               
               
                 XM_006454 
                 CD3G 
                 0.63 
                 0.40 
                 0.75 
                 1.01 
                 6378 
               
               
                 XM_043767 
                 CD3Z 
                 0.43 
                 0.00 
                 0.82 
                 0.77 
                 6379 
               
               
                 XM_056798 
                 CD81 
                 0.50 
                 1.12 
                 0.32 
                 0.00 
                 6380 
               
               
                 M26315 
                 CD8A 
                 1.45 
                 0.00 
                 0.30 
                 1.31 
                 6381 
               
               
                 NM_004931 
                 CD8B1 
                 0.40 
                 0.90 
                 0.50 
                 1.19 
                 6382 
               
               
                 NM_001511 
                 CXCL1 
                 0.09 
                 0.00 
                 0.50 
                 1.34 
                 6385 
               
               
                 XM_057158 
                 ADCY6 
                 1.17 
                 0.00 
                 0.42 
                 1.34 
                 6383 
               
               
                 XM_044428 
                 ICAM2 
                 0.00 
                 1.16 
                 0.50 
                 1.10 
                 6386 
               
               
                 NM_000880 
                 IL7 
                 0.00 
                 1.06 
                 0.74 
                 0.10 
                 6388 
               
               
                 L34657 
                 PECAM-1 
                 0.68 
                 0.39 
                 1.13 
                 0.64 
                 6396 
               
               
                 XM_044882 
                 PTGS1 
                 0.00 
                 1.34 
                 0.52 
                 0.76 
                 6397 
               
               
                 XM_035842 
                 SCYA5 
                 0.60 
                 0.50 
                 0.80 
                 0.99 
                 6401 
               
               
                 NM_021805 
                 SIGIRR 
                 0.00 
                 0.40 
                 0.45 
                 0.66 
                 6402 
               
               
                 XM_057372 
                 TNFRSF5 
                 0.00 
                 0.49 
                 0.59 
                 1.03 
                 6406 
               
               
                 NM_003809 
                 TNFSF12 
                 1.34 
                 0.99 
                 0.53 
                 0.60 
                 6407 
               
               
                   
               
             
          
         
       
     
         [0147]     These characteristic changes can be used for the method according to the present invention.  
         [0148]     In the appended sequence listing, which is part of the present invention, the gene bank accession numbers indicated in tables 2 and 3 (access via internet via http://www.ncbi.nlm.nih.gov/) of the individual sequences are each allocated to one sequence ID.  
       Embodiment 2  
     SIRS  
       [0149]     Study of the gene expression of three patients suffering from SIRS, and one control.  
         [0150]     The gene expression of three patients suffering from SIRS and one control were measured. All patients developed SIRS as described in the criteria according to [4]. The control sample was taken from one patient who was subjected to surgical treatment, but who did not show any SIRS during this stationery treatment. The date of the patients suffering from SIRS and the control are summarized in table 4.  
                                                                   TABLE 4                           Characteristics of the samples of patients and controls                            Apache                           Score   SAPS II       Patient   Gender   Age   Diagnosis   [point]   [point]                    1   male   50   coronary heart   18   36                   disease       2   male   70   caecuM_perforation   19   64       3   male   67   aortic valve   9   21                   insuffiency       1   male   70   fracture of the   1   12                   skull cap                  
 
         [0151]     After the whole blood had been drawn, the total RNA was isolated using the RNAeasy-Kit according to the producer&#39;s (Quiagen) instructions. Subsequently, the cDNA was synthesized from the total RNA by means of reverse transcription with Superscript II RT (Invitrogen), labelled with  33 P according to the producer&#39;s instructions, and hydrolyzed.  
         [0152]     For the hybridization membrane filters of the Deutschen Ressourcenzentrum für Genomforschung GmbH (a German center for genome research) (RZPD) were used. This membrane filter was loaded with about 70,000 human cDNA-molecules.  
         [0153]     The prepared and labelled samples were hybridized with the membrane filter according to the RZPD&#39;s instructions and subsequently washed. The radioactive signals were analyzed after 24 hours of exposition in a phosphorimager.  
         [0000]     Analysis  
         [0154]     The analysis of the gene expression data from the radioactively labelled filters bases on the measurement of the dye intensities in the digitalized picture. This is achieved by the definition of circular areas over all 57600 spot positions, in which the pixel intensities are integrated. The areas are automatically positioned as accurately as possible over the spots by means of the analysis software (AIDA Array Evaluation, raytest Isotopenmessgeräte GmbH).  
         [0155]     In addition to the desired information, namely the amount of nucleic acids bound, contain the detected signals also background signals which are caused by unspecific bindings to the surface of the membrane. In order to eliminate these influences, the background signals are determined in 4608 empty areas of the filter and subtracted as background noise from the hybridization signals.  
         [0156]     In order to render the values of different filters comparable, it is necessary to normalize the data afterwards. Due to the high number of spots on the filter, the mean value of all expression values is set as normalization reference. Further, it is necessary to exclude minor spot signals (lower than 10% of the average expression signal), as these are subject to a percentually high error, and would lead to considerable variations of the results when used later on for calculations.  
         [0157]     The selection of the genes relevant to SIRS bases on the comparison of the gene expression values in a control person not suffering from SIRS compared to the patient suffering from SIRS. The criteria for the grading of the examined genes is the level of the expression ratio. When comparing the genes of the patients with those of the control, the genes, that were significantly overexpressed or underexpressed, respectively, are the interesting ones.  
         [0158]     Table 5 shows that there were 24 genes found in the patient sample, which were significantly overexpressed, if compared with the control sample. Table 6 shows that there were 24 genes found in the patient sample, which were significantly underexpressed, if compared with the control sample. Those results show that the genes listed in table 5 and table 6 correlate with the occurrence of SIRS. Thus, the genes mentioned are leading genes for the diagnosis of SIRS.  
                                         TABLE 5                           Significantly increased transcription activities and       their relative ratio to the control sample in SIRS            GenBank                   SEQUENCE-       Accession No.   HUGO Name   Patient 1:   Patient 2:   Patient 3:   ID               R33626   TFAP2A   57.57   30.43   96.57   6507       N54839   CRSP3   47.17   29.00   63.17   6552       AA010908   LCAT   32.90   15.00   18.60   6561       R59573   TU12B1   85.50   60.50   49.00   6570       R65820   GEF   38.00   45.80   78.00   6594       N30458   NCL   26.57   20.00   17.86   6624       H86783   RINZF   43.33   17.00   31.33   6646       R11676   CDC20   30.75   52.00   55.25   6672       H79834   SLC20A2   16.56   14.33   27.44   6681       H05746   SLC12A5   70.78   20.00   17.22   6685       N21112   ARHGEF12   62.00   14.50   27.00   6693       R71085   PCANAP7   23.00   17.63   21.96   6697       R40287   NIN283   35.00   28.00   28.00   6703       H52708   PDE2A   32.78   14.11   59.22   6723       AF086381   GNPAT   18.94   19.75   25.63   6725       W57892   FN1   23.61   14.67   17.06   6753       H75516   KIN   19.23   17.15   20.00   6761       R59212   MN1   19.65   16.65   18.61   6776       H62284   CMAH   23.40   36.20   32.40   6793       W16423   GCMB   23.83   45.67   21.00   6818       N40557   U5   55.78   20.67   22.11   6826       H52695   DDC   14.80   13.70   22.30   6844       R68244   HMG14   15.81   23.19   27.56   6865       R34679   ITGB8   19.20   32.00   79.20   6874                  
 
         [0159]    
       
         
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                   
               
               
                 Significantly reduced transcription activities and 
               
               
                 their relative ratio to the control sample in SIRS 
               
             
          
           
               
                 GenBank 
                   
                   
                   
                   
                 SEQUENCE- 
               
               
                 Accession No. 
                 HUGO Name 
                 Patient 1: 
                 Patient 2: 
                 Patient 3: 
                 ID 
               
               
                   
               
               
                 H18595 
                 RPL10A 
                 0.03 
                 0.07 
                 0.15 
                 6553 
               
               
                 N90220 
                 DGUOK 
                 0.04 
                 0.07 
                 0.12 
                 6574 
               
               
                 R19651 
                 H19 
                 0.09 
                 0.07 
                 0.19 
                 6701 
               
               
                 R52108 
                 UBE2D2 
                 0.13 
                 0.07 
                 0.02 
                 6741 
               
               
                 R83836 
                 LYN 
                 0.07 
                 0.03 
                 0.18 
                 6759 
               
               
                 H04648 
                 CSF2RB 
                 0.06 
                 0.07 
                 0.13 
                 6767 
               
               
                 H27730 
                 PPP2R1B 
                 0.09 
                 0.07 
                 0.16 
                 6788 
               
               
                 N70020 
                 PRO2822 
                 0.10 
                 0.04 
                 0.11 
                 6794 
               
               
                 N52437 
                 CHI3L2 
                 0.07 
                 0.08 
                 0.16 
                 6812 
               
               
                 W96179 
                 GCLM 
                 0.04 
                 0.01 
                 0.19 
                 6822 
               
               
                 H42506 
                 GABARAP 
                 0.08 
                 0.03 
                 0.17 
                 6842 
               
               
                 H66258 
                 SCP2 
                 0.10 
                 0.05 
                 0.21 
                 6846 
               
               
                 N38985 
                 RAP140 
                 0.10 
                 0.06 
                 0.21 
                 6896 
               
               
                 N73912 
                 TMP21 
                 0.09 
                 0.07 
                 0.08 
                 6905 
               
               
                 N51024 
                 TEGT 
                 0.08 
                 0.06 
                 0.07 
                 6909 
               
               
                 R99466 
                 EEF1A1 
                 0.07 
                 0.02 
                 0.14 
                 7008 
               
               
                 R14080 
                 CAMLG 
                 0.11 
                 0.02 
                 0.18 
                 7034 
               
               
                 W93782 
                 XPC 
                 0.12 
                 0.05 
                 0.21 
                 7036 
               
               
                 N91584 
                 RPS6 
                 0.06 
                 0.05 
                 0.12 
                 7353 
               
               
                 W52982 
                 PIG7 
                 0.05 
                 0.07 
                 0.10 
                 7412 
               
               
                 AA033725 
                 KLF8 
                 0.06 
                 0.08 
                 0.19 
                 7535 
               
               
                 N20406 
                 SRP14 
                 0.10 
                 0.04 
                 0.16 
                 7565 
               
               
                 T83104 
                 TAF2F 
                 0.02 
                 0.05 
                 0.12 
                 7630 
               
               
                 H79277 
                 CASP8 
                 0.12 
                 0.06 
                 0.13 
                 7677 
               
               
                   
               
             
          
         
       
     
         [0160]     These characteristic changes can be used for the method according to the present invention.  
         [0161]     In the appended sequence listing (SEQ. ID No: 6373 to SEQ. ID No: 10540), which is part of the present invention, the gene bank accession numbers indicated in tables 5 and 6 (access via internet via http://www.ncbi.nlm.nih.gov/) of the individual sequences are each allocated to one sequence ID.  
       Embodiment 3  
     Sepsis  
       [0162]     Study of the gene expression of one patient suffering from an early sepsis and one control sample.  
         [0163]     The gene expression of one case of an early sepsis and one control sample were measured. The patient&#39;s data are summarized in table 7.  
                                                                                                           TABLE 7                           Data of the samples of patients and controls                                            Apache                   Gen-   Age   Weight/       Intercurrent           Score   SAPS II   Selection of           der   [a]   Height   Main diagnosis   diagnosis   Operations   Indication   [point]   [point]   clinical data                        Patient   male   70   78 kg/   septic shock   intestine-,   1. Anastomotic-   Sepsis/   19   64   temperature: 35.2° C.                   178 cm   after caecum   instable   and sigma re-   septic           heart rate: 97/min                       perforation and   sternum   resection, rectum   shock           MAP 1: 62 mmHg;                       post operative       dead end               art. PH: 7.29                       anastomotic leak       blockage               Na: 135 mmol/l;                               2. Punctation               Creatine: 757 mmol/l;                               tracheotomy               Cholesterol: -                               (Griggs)               Breathing rate: 16/min                               3. re-wiring               Syst. BP: 105 mmHg;                               4. subtotal               Haematocrit: 33%                               hemiclolectomy               Total number of                               right side               leucocytes: 13100                               5. definitive               Urea: 19 mmol/l;                               ileostomy               Diast. BP: 40 mmHg;                               surgery               PaO2: 12.3 kPa;                                               K: 4.2 mmol/l;                                               Bilirubin: 15.1 mmol/l;       Control   male   35   90 kg/   Fracture of the   small hygroma   1. Craniotomy   Intacranial   1   12   Temp: 38.8° C.                   180 cm   skull, scalp   on both sides   and definite   bleeding           heart rate: 84/min                       haematoma       haemostasis               MAP 1: 72 mmHg;                                               art. PH: 7.42/l                                               Na: 140 mmol                                               Creatine: 56 μmol/l;                                               Breathing rate: 13/min                                               Syst. BD: 107 mmHg;                                               Haematocrit: 37%                                               HCO3: 28.2 mmol/l;                                               Total number of                                               leucocytes: 12600                                               Urea: 4.7 mmol/l;                                               Diast. Syst. BD: 54                                               mmHg;                                               PaO2: 10.9 kPa;                                               K: 3.8 mmol/l;                                               Bilirubin: 13.4 mmol/l;                  
 
         [0164]     After the whole blood had been drawn, the total RNA was isolated using RNAeasy according to the producer&#39;s (Quiagen) instructions. Subsequently, the cDNA was synthesized from the total RNA by means of reverse transcriptions with Superscript II RT (Invitrogen), labelled with  33 P, according to the producer&#39;s instructions, and hydrolyzed.  
         [0165]     For the hybridization membrane filters of the Deutschen Ressourcenzentrum für Genomforschung GmbH (RZPD) were used. This membrane filter was loaded with about 70,000 humane cDNA-molecules.  
         [0166]     The prepared and labelled samples were hybridized by means of the membrane filter according to the RZPD&#39;s instructions and subsequently washed. The radioactive signals were analyzed after 24 hours of exposition in a phosphorimager.  
         [0167]     The expression ratios of the samples of the patients and the control were calculated from the signal intensities using the AIDA Array Evaluation software.  
         [0168]     Table 8 shows that 230 genes of the patient sample were found, which were significantly overexpressed (expression ratios between 13.67 and 98.33), if compared with the control sample. Table 3 further shows that 206 genes of the patient sample were found, which were significantly underexpressed (expression ratios between 0.01 and 0.09), if compared with the control sample. Those results show that the genes listed in table 2 and table 3 correlate with the occurrence of SIRS. Thus, the genes mentioned are leading genes for the diagnosis of an early sepsis.  
                                                   TABLE 8                           Expression ratio of overexpressed genes       of samples of patients and controls            GenBank                   Gene Bank       Expression ratio of       Accession       overexpressed genes   SEQUENCE-       No.   HUGO Name   compared to control   ID                        FLJ20623   90.13   325       AI272878   FGF20   73.48   268       AI218453   FLJ22419   48.8   294       AI473374   SPAM1   42.63   235       AI301232   PRG4   36.79   262       AI452559   FLJ13710   32   240       AI339669   FLJ21458   31   248       AI142427   CGRP-RCP   30   331       AA505969   LOC56994   26.67   486       AI333774   AGM1   26.19   251       W86875   PSEN1   25.66   903       AI591043   NR2E3   25   196       AI128812   RBM9   23.56   324       AA453019   FLJ21924   23.07   672       AI690321   KCNK15   22.71   134       AA918208   ADAM5   21.83   363       AI344681   ABCA1   21.42   259       AI654100   KIAA0610   21.04   168       AI086719   FLJ12604   20.95   326       AA453038   LOC63928   20.74   671       AI740697   SP3   20.5   114       AI332438   KIAA1033   20.17   253       AI734941   MSR1   19.93   116       AA541644   PRV1   19.51   489       AA513806   C5ORF3   19.3   485       AI381513   B4GALT7   18.81   273       AI671360   SIM1   18.55   154       AI624830   SAGE   17.54   187       AI001846   KIAA0480   17.54   358       AA504336   TRAP95   17.25   495       AI142901   IMPACT   17.15   330       AI077481   SEMA5B   17.13   327       H41851   TNFRSF12   17.05   1511       AI160574   FLJ23231   17   314       AI033829   KIF13B   16.59   339       AI554655   HLALS   16.59   219       AI074113   LOC51095   16.4   328       AA992716   KIAA1377   16.14   348       AI382219   SETBP1   16.08   272       AI469528   KIAA1517   15.89   232       AI090008   NFYB   15.76   349       AI203498   WRN   15.72   310       AI832179   HPGD   15.66   65       AI278521   SPRR3   15.61   265       AA909201   FLJ23129   15.12   361       AI383932   ZNF214   14.98   269       AA455096   MDM1   14.9   652       AA953859   NOL4   14.68   363       R56800   GDF1   14.67   1755       AI676097   FCER1A   14.54   151       AI380703   KIAA1268   14.51   275       AI832086   RTKN   14.51   66       AI125328   FLJ22490   14.33   317       AI056693   LOC57115   14.3   329                  
 
         [0169]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                   
               
               
                 Expression ratio of underexpressed genes 
               
               
                 of samples of patients and controls 
               
             
          
           
               
                 GenBank 
                   
                 Expression ratio of 
                   
               
               
                 Accession 
                   
                 underexpressed genes 
                 SEQUENCE- 
               
               
                 No. 
                 Hugo Name 
                 compared to control 
                 ID 
               
               
                   
               
             
          
           
               
                 R15296 
                 C9ORF9 
                 0.01 
                 2050 
               
               
                 AA609149 
                 FLJ10058 
                 0.01 
                 375 
               
               
                 AI566451 
                 KAI1 
                 0.01 
                 211 
               
               
                 AI334246 
                 PDCD7 
                 0.01 
                 250 
               
               
                 H38679 
                 NXPH3 
                 0.01 
                 1477 
               
               
                 AI696866 
                 KIAA1430 
                 0.01 
                 130 
               
               
                 AI922915 
                 FLJ00012 
                 0.01 
                 23 
               
               
                 AI889612 
                 KPNA6 
                 0.01 
                 46 
               
               
                 AI921695 
                 FLJ23556 
                 0.02 
                 26 
               
               
                 AA410933 
                 HRH1 
                 0.02 
                 764 
               
               
                 AA705423 
                 LOC57799 
                 0.02 
                 383 
               
               
                 AI206507 
                 RAD54B 
                 0.02 
                 298 
               
               
                 AI921327 
                 MED6 
                 0.02 
                 28 
               
               
                 AI682701 
                 VNN1 
                 0.02 
                 146 
               
               
                 H82822 
                 METAP2 
                 0.02 
                 1352 
               
               
                 AI890612 
                 MAGE1 
                 0.02 
                 42 
               
               
                 AI262169 
                 ALDOB 
                 0.02 
                 257 
               
               
                 H44908 
                 C21ORF51 
                 0.02 
                 1502 
               
               
                 AI572407 
                 FLJ22833 
                 0.02 
                 203 
               
               
                 AI924869 
                 STX4A 
                 0.02 
                 19 
               
               
                 AI925556 
                 AF140225 
                 0.02 
                 12 
               
               
                 AI798388 
                 KIAA0912 
                 0.03 
                 95 
               
               
                 AI623978 
                 SCEL 
                 0.03 
                 188 
               
               
                 AI889598 
                 MLYCD 
                 0.03 
                 47 
               
               
                 AI889648 
                 PAWR 
                 0.03 
                 45 
               
               
                 AI431323 
                 AREG 
                 0.03 
                 237 
               
               
                 AA446611 
                 CDH6 
                 0.03 
                 706 
               
               
                 AI697365 
                 P53DINP1 
                 0.03 
                 129 
               
               
                 H82767 
                 VAMP3 
                 0.03 
                 1353 
               
               
                 AI688916 
                 FLJ10933 
                 0.03 
                 137 
               
               
                 AI888660 
                 FLJ11506 
                 0.03 
                 51 
               
               
                 AI890314 
                 RAB6B 
                 0.03 
                 43 
               
               
                 AI653893 
                 LAMA5 
                 0.03 
                 169 
               
               
                 R89811 
                 HGFAC 
                 0.03 
                 1462 
               
               
                 AI863022 
                 MAGEA4 
                 0.04 
                 59 
               
               
                 AA749151 
                 XPOT 
                 0.04 
                 378 
               
               
                 AI355007 
                 ITPKB 
                 0.04 
                 246 
               
               
                 AI582909 
                 MESDC2 
                 0.04 
                 201 
               
               
                 AI832016 
                 APOL1 
                 0.04 
                 67 
               
               
                 H11827 
                 THOP1 
                 0.04 
                 1597 
               
               
                 AI560205 
                 KIAA1841 
                 0.04 
                 216 
               
               
                 AA503092 
                 UMPH1 
                 0.04 
                 490 
               
               
                 AI932616 
                 FLJ22294 
                 0.04 
                 5 
               
               
                 AI799137 
                 FLJ11274 
                 0.04 
                 93 
               
               
                 AI686838 
                 SARDH 
                 0.04 
                 142 
               
               
                 AI623132 
                 SREC 
                 0.04 
                 189 
               
               
                 R96713 
                 DKFZP434A0131 
                 0.04 
                 1442 
               
               
                 AI674926 
                 LBC 
                 0.04 
                 152 
               
               
                 AI886302 
                 HRI 
                 0.04 
                 53 
               
               
                 AI434650 
                 MGC2560 
                 0.04 
                 238 
               
               
                 AI631380 
                 GNG4 
                 0.04 
                 180 
               
               
                 AA508868 
                 ORC6L 
                 0.04 
                 491 
               
               
                 AI620374 
                 HP1-BP74 
                 0.04 
                 190 
               
               
                 AI679115 
                 KIAA1353 
                 0.04 
                 148 
               
               
                 AA652703 
                 MRPL49 
                 0.04 
                 386 
               
               
                 AI355775 
                 CDK3 
                 0.04 
                 245 
               
               
                   
               
             
          
         
       
     
         [0170]     These characteristic alterations can be used in particular for the method of the present invention.  
         [0171]     In the appended sequence listing, which is part of the present invention, the gene bank accession numbers (access via internet via http://www.ncbi.nlm.nih.gov/) indicated in tables 8 and 9 of the individual sequences are each allocated to one sequence ID.  
         [0000]     Implementation:  
         [0172]     Preparation of RNA. The conditioned media were removed from the culture flasks and the adherent cells were lysed directly in the culture flasks using TRIzol-reagent (GIBCO/BRL) according to the producer&#39;s instructions. One deproteinization cycle was carried out and afterwards, the RNA was precipitated by adding isopropyl alcohol, afterwards rinsed with ethyl alcohol, and again solved in 200 μl RNA-save resuspension solution (Ambion, Austin, Tex.). The RNA preparations were degraded with 0.1 units/μl DNase I, in DNase 1 buffer from CLONTECH. Additionally, proteins were removed from the RNA units in an alcohol mixture comprising phenol, chloroform and isoamyl alcohol, precipitated by adding ethyl alcohol, and solved in 50-100 μl RNA-save resuspension solution. The RNA concentration was spectro-photometrically determined, provided that 1A 260  corresponds to a concentration of 40 μg/ml. The samples were adapted to a final concentration of 1 mg/ml und stored at 80° C. No signs of deterioration of quality were observed. By means of agarose electrophoresis it was evaluated whether the RNA preparations were complete (i.e. they were not decayed into their components), here, RNA-standards (GIBCO/BRL) were used. Each of the preparations described herein contained intact RNA the 28S-, 18S- and 5S-bands of which were clearly detectable (data not given). No recognizable differences between healthy and infectious cells were determined with regard to the electrophoretically determined RNA samples.  
         [0173]     Preparation of radioactively labelled cDNA-samples and hybridizing by means of DNA arrays. The cDNA-synthesis was carried out according to the producer&#39;s instructions using gene specific primer (CLONTECH) and [ 32 P]-dATP with Moloney Murine Leukemea Virus Reverse Transkriptase (SuperScript II, GIBCO/BRL). For the cDNA-synthesis, the same amounts of RNA (5 μg) were used from each sample.  
         [0000]     Alternative  
         [0174]     RNA was extracted from the tissue samples by means of guanidinium thiocyanate and afterwards centrifuged in CsCl as described [19]. The RNA was extracted according to the producer&#39;s instructions from the cell lines with RNAzol (Biotex Laboratories, Houston). The poly(A) RNA was isolated from 500 μg RNA by means of DynaBeads (Dynal, Oslo), as described in the producer&#39;s recommendations.  
         [0175]     The differences in the gene expression were examined using Atlas Array membranes (CLONTECH). A first short step was the transcription of 1 μg RNA of each cell line in [− 32 P]dATP-labelled cDNA at a time.  
         [0000]     Analysis  
         [0176]     The analysis of the gene expression data from the radioactively labelled filters bases on the measurement of the dye intensities in the digitalized picture. This is achieved by the definition of circular areas over all 57600 spot positions, in which the pixel intensities are integrated. The areas are automatically positioned as accurately as possible over the spots by means of the analysis software (AIDA Array Evaluation, raytest Isotopenmessgeräte GmbH).  
         [0177]     In addition to the desired information, namely the amount of nucleic acids bound, contain the detected signals also background signals which are caused by unspecific bindings to the surface of the membrane. In order to eliminate these influences, the background signals are determined in 4608 empty areas of the filters and subtracted as background noise from the hybridization signals.  
         [0178]     It is possible to distinguish between punctual signals that are caused on the filter by dust particles or other disturbances instead of binding of nucleic acids, and real spots, due to their irregular form, and the punctual signals are excepted from further analysis.  
         [0179]     In order to render the values of different filters comparable, it was necessary to normalize the data afterwards. Due to the high number of spots on the filter, the mean value of all expression values is set as normalization reference. Further, it is necessary to exclude minor spot signals (lower than 10% of the average expression signal), as these are subject to a percentually high error, and would lead to considerable variations of the results when used later on for calculations.  
         [0180]     The selection of the genes relevant to SIRS/sepsis bases on the comparison of the gene expression values in a control person without SIRS/sepsis compared to one patient at a time suffering from sepsis/SIRS. The criteria for the grading of the examined genes is the level of the expression ratio. The interesting genes are those which were most overexpressed or underexpressed, respectively, in the patients compared with the control.  
       Embodiment 4  
     Sepsis  
       [0181]     Study of the protein expression of one patient suffering from sepsis and one control sample.  
         [0182]     The protein expression of one case of sepsis and one control sample were measured. The patients&#39; data are summarized in table 10.  
                                                                                                                       TABLE 10                       Data of the samples of patients and controls                                        Age       Main               Gender   [a]   Weight/Height   diagnosis   Intercurrent diagnosis                        Control   female   21   62 kg/167 cm   cranio-   Generalized cerebral oedema, brain stem contusion,                       cerebral-   fracture of the lateral orbital pillar, fracture                       trauma   wall left side, lateral fracture of the nasal                           sceleton, bleeding into the right side ventricle,                           free air intracraniellfrontally left side, ethmoid                           bone fracture, fracture of the front pelvic ring with                           impression and dislocation of the fragments, fracture                           of the massa lateralis of the OS sacrum right side                           in the heigh of S1/S2, clavikular fracture left side       Patient   male   59   70 kg/175 cm   septic shock   pleural effusions on both sides, multi organ failure,                       after   multiple necrosis of the acra and pretibial on both                       perforation of   sides, arterial microembolism, arterial thrombosis,                       one ulcus   secundary thrombocytopenia, acute kidney failure                       pylori and                       subsequent 4                       quadrant                       peritonitis                                Apache                           Score   SAPS II           Operations   Indication   [point]   [point]   Selection of clinical data                        Control   none   not applicable   21   —   temperature: 35.3° c.                           heart rate: 146/min                           map 1: 68 mmhg; art. ph: 7.48                           na: 145 mmol/l; ceratine: 52 μmol/l;                           syst. bp: 94 mmhg; diast. bp: 56 mmhg;                           haematocrit: 0.26%                           total number of leucocytes: 9200                           urea: 7.1 mmol/l;                           k: 5 mmol/l;                           bilirubin: 11.1 mmol/l;       Patient   relaparotomy,   septic shock   28   74   temperature: 37.7° c.           lavage, and partial               heart rate: 139/min           resection of the               map 1: 64 mmhg; art. ph: 7.15           omentum               na: 142 mmol/l, ceratine: 187 mmol/l;                           breathing rate: 19/min                           syst. bp: 99 mmhg; diast. bp: 49                           mmhg; haematocrit: 24%                           hco3: 13.7 mmol/l, total number of                           leucocytes: 5200                           urea: 27.6 mmol/l;                           pao2!: 13.2 kpa, k: 5.3 mmol/l;                           bilirubin: 33.9 mmol/l;                           urine: 110 ml, 14 h                  
 
         [0183]     Whole blood was drawn and inserted into a serum tube and centrifugation (5500 rcf; 10 min; 4° C.) was carried out. The supernatant of serum was transferred into cryo tubes immediately upon centrifugation and stored at −35° C.  
         [0184]     To downgrade the albumin, the serum was treated with Affi-Gel Blue Affinity Chromatography Gel for Enzyme and Blood Protein Purifications (Bio-Rad) according to the producer&#39;s instructions. To avoid undesired interactions of protein and matrix, the equilibration- and binding buffer were added 400 mM NaCl.  
         [0185]     Non-binding proteins were collected and precipitated with methanol and chloroform according to the protocol of Wessel and Flügge (Anal. Biochem. 1984 April; 138(1): 141-3). 250 microgram of precipitated serum protein were added to a solution consisting of 8M urea; 2.0 M thiourea; 4% CHAPS; 65 mM DTT and 0.4% (w/v) Bio-Lytes 3/10 (Bio-Rad) and subjected to an isoelectric focusing as well as a subsequent SDS-PAGE.  
       SDS-PAGE  
       [0186]     K4 in  FIG. 1  and in  FIG. 2  is the acute phase protein transthyretin (TTR; P02766, SEQ. ID 6241, SEQ. ID 6242) and K5 and K6 are the vitamin D-binding protein (DBP; P02774, SEQ. ID 1554, SEQ. ID 1555).  
         [0187]     The gels can be produced as follows (Cibacron FT, W1-W3, 400 mM NaCl, IEF pH 3-10, Coomassie):  
         [0188]     250 microgram of precipitated serum protein were added to a solution consisting of 8M urea; 2.0 M thiourea; 4% CHAPS; 65 mM DTT and 0.4% (w/v) Bio-Lytes 3/10 (Bio-Rad) and subjected to an isoelectric focusing as well as a subsequent SDS-PAGE.  
         [0189]     The prepared 2-dimensional gels were colored with Coomassie Brilliant Blau G-250 and differentially expressed proteins were identified by mass spectroscopy.  
         [0190]     A comparing analysis shows ( FIG. 1 ,  FIG. 2 =that the acute phase protein transthyretin (TTR; P02766, SEQ. ID: 6241, SEQ. ID 6242), as well as the vitamin D-binding protein (DBP; P02774, SEQ. ID 1554, SEQ. ID 1555) are less expressed by the sepsis patient, if compared with the control patient.  
         [0191]     These results clearly indicate that the protein expression or the protein composition, respectively, of serum and plasma change in the course of the disease.  
       Embodiment 5  
     Severe Sepsis  
       [0192]     Studies of differential gene expression with patients suffering from severe sepsis.  
         [0193]     Whole blood samples of patients who were under the care of a surgical intensive care unit were examined for the measurement of the differential gene expression in connection with severe sepsis.  
         [0194]     Control samples were whole blood samples of the patients that were drawn after an uncomplicated neurosurgical operation. The patients were treated on the same intensive care unit. No one of these patients developed an infection and/or showed clinical signs of a generalized inflammatory reaction (defined according to the SIRS-criteria [4]) during the whole time of stationary treatment.  
         [0195]     Additionally, whole blood samples were drawn from six male and two female patients (patients&#39; samples). In the time period of 24 hours before the whole blood was drawn, each of these patients developed a new severe sepsis with organ dysfunction. A range of characteristics of the patients suffering from severe sepsis are shown in table 1. Information concerning the age, gender, the cause of the severe sepsis (see diagnosis) as well as concerning the clinical severity, measured with the APACHE-II- and SOFA-Scores (in points each), that are well documented in clinical literature, is given. Equally, the plasma protein levels of procalcitonin (PCT), a new kind of sepsis label, are given, as well as the individual survival conditions.  
                                                                                   TABLE 11                           Data of the group of patients                            Apache II   SOFA                           Classification   Score   Score   PCT   survival       Age   Gender   Diagnosis   according to [4]   [points]   [points]   [ng/ml]   conditions                    68   female   peritonitis   severe   17   4   269   died                   sepsis/       39   male   ARDS   septic shock   17   11   0.39   died       36   male   peritonitis   septic shock   11   7   9.77   survived       80   male   peritonitis   severe   28   4   23.61   survived                   sepsis       32   male   bacterial   septic shock   21   7   1.69   survived               pancreatitis       73   male   ARDS   septic shock   16   14   9.96   died       67   male   ARDS   septic shock   24   12   12.88   survived       76   female   peritonitis   septic shock   30   11   4.19   died                  
 
         [0196]     After the whole blood had been drawn, the total RNA was isolated using the PAXGene Blood RNS Kit according to the producer&#39;s (Qiagen) instructions. Subsequently, the cDNA was synthesized from the total RNA by means of reverse transcription with Superscript II RT (Invitrogen) according to the producer&#39;s instructions, labelled with aminoallyl-dUTP and succinimidylester of the fluorescent dyes Cy3 and Cy5 (Amersham), and hydrolyzed.  
         [0197]     The microarrays (Lab-Arraytor human 500-1 cDNA) of the company SIRS-Lab GmbH were used for the hybridization. These micorarrays are loaded with 340 human cDNA-molecules. The 340 human cDNA-molecules are 3-fold immobilized in three subarrays on each microarray.  
         [0198]     The prepared and labelled samples were hybridized with the microarrays according to the producer&#39;s instructions and subsequently washed. The fluorescence signals of the hybridized molecules were measured by means of a scanner (AXON 4000B).  
         [0000]     Analysis  
         [0199]     One test was analyzed by means of scanned pictures of the microarrays after hybridization. The mean intensity value of the detected spots were defined as the measured expression value of the corresponding gene. Spots were automatically identified by means of picture analysis and their homogeneity was checked. The analysis was controlled manually. The detected signals comprise not only the desired information, namely the amount of nucleic acids bound, but also background signals which are caused by unspecific bindings to the surface of the membrane. The definition of the signals of the background rendered an optimum differentiation between spots and the surface of the chip possible, which surface also showed color effects. For the analysis of the microarrays blank spots were chosen as background. The mean expression value of the chosen blank spots within one block (of 14 times 14 spots) was subtracted from the expression values of the gene spots (in the corresponding block).  
         [0200]     It was possible to distinguish between punctual signals that were caused on the filter by dust particles or other disturbances instead of bindings of nucleic acids, and real spots, due to their irregular form, and the punctual signals were excepted from further analysis.  
         [0201]     In order to render the values between the 3 subarrays and between different microarrays comparable, it was necessary to normalize the data afterwards. Due to the high number of spots on the microarray, the mean value of all expression values was set as normalization reference. The mean expression per gene was calculated by choosing the two (from three) repetitions that were closest to each other.  
         [0202]     The expression ratios of the samples of the patients and the control were calculated from the signal intensities using the AIDA Array Evaluation software. The criterion for the grading of the examined genes was the level of the expression ratio. The interesting genes were those which were most overexpressed or underexpressed, respectively, compared with the control samples.  
         [0203]     Table 12 shows that 41 genes of the patient sample were found, which were significantly overexpressed, if compared with the control sample. Table 13 shows that 89 genes of the patient sample were found, which were significantly underexpressed, if compared with the control sample. Those results show that the genes listed in table 12 and table 13 correlate with the occurrence of a severe sepsis. Furthermore, these results correlate with the clinical classification according to [4] as well as patients&#39; PCT-concentrations, that are typical for the occurrence of a severe sepsis [23]. Thus, the gene activities of the genes mentioned are labels for the diagnosis of a severe sepsis.  
                                                   TABLE 12                           Expression ratio of overexpressed genes       of samples of patients and controls            GenBank       Expression ratio of           Accession       overexpressed genes   SEQUENCE-       No.   HUGO Name   compared to control   ID                    XM_086400   S100A8   4.4   6243       XM_001682   S100A12   3.03   6244       NM_002619   PF4   2.21   6245       NM_002704   PPBP   1.66   6246       NM_001101   ACTB   1.65   6247       NM_001013   RPS9   1.61   6248       XM_057445   SELP   1.61   6249       BC018761   IGKC   1.53   6250       XM_030906   TGFB1   1.51   6251       NM_001760   CCND3   1.48   6252       XM_035922   IL11   1.28   6253       XM_039625   DUSP10   1.17   6254       XM_002762   TNFAIP6   1.17   6255       XM_015396   ALOX5AP   1.15   6256       NM_003823   TNFRSF6B   1.15   6257       XM_029300   DPP4   1.15   6258       NM_001562   IL18   1.14   6259       NM_005037   PPARG   1.11   6260       M90746   FCGR3B   1.07   6261       NM_001315   MAPK14   0.99   6262       BC001506   CD59   0.88   6263       XM_042018   BSG   0.88   6264       XM_010177   DUSP9   0.87   6265       BC013992   MAPK3   0.84   6266       NM_001560   IL13RA1   0.82   6267       NM_004555   NFATC3   0.74   6268       NM_001154   ANXA5   0.73   6269       NM_001310   CREBL2   0.7   6270       XM_036107   ITGB2   0.65   6271       XM_009064   JUNB   0.65   6272       NM_001774   CD37   0.62   6273       XM_049849   TNFRSF14   0.6   6274       NM_003327   TNFRSF4   0.57   6275       BC001374   CD151   0.56   6276       XM_051958   ALOX5   0.56   6277       NM_021805   SIGIRR   0.5   6278       NM_017526   HSOBRGR   0.48   6279       XM_011780   DAPK1   0.46   6280       NM_006017   PROML1   0.44   6281       D49410   IL3RA   0.43   6372       XM_027885   RPL13A   0.33   6282                  
 
         [0204]    
       
         
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 13 
               
             
             
               
                   
               
               
                   
               
               
                 Expression ratio of underexpressed genes 
               
               
                 of samples of patients and controls 
               
             
          
           
               
                 GenBank 
                   
                 Expression ratio of 
                   
               
               
                 Accession 
                   
                 underexpressed genes 
                 SEQUENCE- 
               
               
                 No. 
                 HUGO Name 
                 compared to control 
                 ID 
               
               
                   
               
             
          
           
               
                 NM_007289 
                 MME 
                 −2.11 
                 6283 
               
               
                 XM_008411 
                 SCYA13 
                 −2.06 
                 6284 
               
               
                 XM_055188 
                 ENG 
                 −2.01 
                 6285 
               
               
                 NM_021073 
                 BMP5 
                 −1.99 
                 6286 
               
               
                 XM_007417 
                 TGFB3 
                 −1.93 
                 6287 
               
               
                 NM_001495 
                 GFRA2 
                 −1.88 
                 6288 
               
               
                 XM_009475 
                 AHCY 
                 −1.86 
                 6289 
               
               
                 XM_006738 
                 CD36L1 
                 −1.86 
                 6290 
               
               
                 NM_001772 
                 CD33 
                 −1.86 
                 6291 
               
               
                 NM_057158 
                 DUSP4 
                 −1.83 
                 6292 
               
               
                 XM_058179 
                 CD244 
                 −1.77 
                 6293 
               
               
                 NM_001770 
                 CD19 
                 −1.75 
                 6294 
               
               
                 NM_004931 
                 CD8B1 
                 −1.73 
                 6295 
               
               
                 XM_006454 
                 CD3G 
                 −1.71 
                 6296 
               
               
                 XM_041847 
                 TNF 
                 −1.65 
                 6297 
               
               
                 NM_145319 
                 MAP3K6 
                 −1.62 
                 6298 
               
               
                 XM_045985 
                 ITGA2B 
                 −1.61 
                 6299 
               
               
                 XM_055756 
                 TIMP1 
                 −1.61 
                 6300 
               
               
                 NM_004740 
                 TIAF1 
                 −1.61 
                 6301 
               
               
                 XM_008432 
                 ITGA3 
                 −1.57 
                 6302 
               
               
                 XM_034770 
                 PAFAH1B1 
                 −1.56 
                 6303 
               
               
                 NM_014326 
                 DAPK2 
                 −1.55 
                 6304 
               
               
                 XM_043864 
                 PIK3R1 
                 −1.49 
                 6305 
               
               
                 U54994 
                 CCR5 
                 −1.49 
                 6306 
               
               
                 NM_004089 
                 DSIPI 
                 −1.49 
                 6307 
               
               
                 XM_037260 
                 F2R 
                 −1.45 
                 6308 
               
               
                 NM_172217 
                 IL16 
                 −1.45 
                 6309 
               
               
                 AF244129 
                 LY9 
                 −1.45 
                 6310 
               
               
                 NM_003775 
                 EDG6 
                 −1.43 
                 6311 
               
               
                 NM_001781 
                 CD69 
                 −1.41 
                 6312 
               
               
                 NM_019846 
                 CCL28 
                 −1.39 
                 6313 
               
               
                 NM_001511 
                 CXCL1 
                 −1.38 
                 6314 
               
               
                 NM_006505 
                 PVR 
                 −1.33 
                 6315 
               
               
                 NM_000075 
                 CDK4 
                 −1.33 
                 6316 
               
               
                 XM_042066 
                 MAP3K1 
                 −1.32 
                 6317 
               
               
                 NM_003242 
                 TGFBR2 
                 −1.31 
                 6318 
               
               
                 NM_003874 
                 CD84 
                 −1.31 
                 6319 
               
               
                 XM_033972 
                 ATF6 
                 −1.3 
                 6320 
               
               
                 XM_001840 
                 PLA2G2A 
                 −1.3 
                 6321 
               
               
                 NM_018310 
                 BRF2 
                 −1.29 
                 6322 
               
               
                 AF212365 
                 IL17BR 
                 −1.25 
                 6323 
               
               
                 XM_056798 
                 CD81 
                 −1.25 
                 6324 
               
               
                 NM_000061 
                 BTK 
                 −1.24 
                 6325 
               
               
                 XM_001472 
                 JUN 
                 −1.23 
                 6326 
               
               
                 XM_007258 
                 TNFAIP2 
                 −1.23 
                 6327 
               
               
                 XM_048555 
                 IFNAR2 
                 −1.23 
                 6328 
               
               
                 XM_041060 
                 FOS 
                 −1.23 
                 6329 
               
               
                 XM_056556 
                 TNFSF7 
                 −1.23 
                 6330 
               
               
                 XM_016747 
                 LTBP1 
                 −1.22 
                 6331 
               
               
                 XM_006953 
                 TNFRSF7 
                 −1.21 
                 6332 
               
               
                 NM_015927 
                 TGFB1I1 
                 −1.19 
                 6333 
               
               
                 XM_010807 
                 INHBB 
                 −1.16 
                 6334 
               
               
                 NM_002184 
                 IL6ST 
                 −1.14 
                 6335 
               
               
                 XM_008570 
                 VAMP2 
                 −1.13 
                 6336 
               
               
                 NM_006856 
                 ATF7 
                 −1.1 
                 6337 
               
               
                 NM_000674 
                 ADORA1 
                 −1.09 
                 6338 
               
               
                 NM_000173 
                 GP1BA 
                 −1.08 
                 6339 
               
               
                 XM_048068 
                 SCYD1 
                 −1.07 
                 6340 
               
               
                 NM_022162 
                 CARD15 
                 −1.07 
                 6341 
               
               
                 NM_001199 
                 BMP1 
                 −1.02 
                 6342 
               
               
                 NM_000960 
                 PTGIR 
                 −1.01 
                 6343 
               
               
                 XM_012039 
                 FUT4 
                 −0.99 
                 6344 
               
               
                 XM_034166 
                 NOS2A 
                 −0.99 
                 6345 
               
               
                 NM_003188 
                 MAP3K7 
                 −0.98 
                 6346 
               
               
                 NM_006609 
                 MAP3K2 
                 −0.98 
                 6347 
               
               
                 XM_027358 
                 PCMT1 
                 −0.95 
                 6348 
               
               
                 XM_007189 
                 FOXO1A 
                 −0.93 
                 6349 
               
               
                 XM_030523 
                 MAP3K8 
                 −0.92 
                 6350 
               
               
                 XM_002923 
                 CCR2 
                 −0.88 
                 6351 
               
               
                 XM_054837 
                 TNFRSF1B 
                 −0.87 
                 6352 
               
               
                 NM_000634 
                 IL8RA 
                 −0.87 
                 6353 
               
               
                 NM_004590 
                 CCL16 
                 −0.86 
                 6354 
               
               
                 XM_012717 
                 CSNK1D 
                 −0.86 
                 6355 
               
               
                 XM_012649 
                 SCYA7 
                 −0.84 
                 6356 
               
               
                 XM_008679 
                 TP53 
                 −0.84 
                 6357 
               
               
                 XM_030509 
                 PTGIS 
                 −0.83 
                 6358 
               
               
                 XM_039086 
                 CDW52 
                 −0.82 
                 6359 
               
               
                 XM_027978 
                 CFLAR 
                 −0.81 
                 6360 
               
               
                 NM_005343 
                 HRAS 
                 −0.79 
                 6361 
               
               
                 XM_043574 
                 DAP3 
                 −0.78 
                 6362 
               
               
                 NM_002188 
                 IL13 
                 −0.77 
                 6363 
               
               
                 XM_055699 
                 ENTPD1 
                 −0.72 
                 6364 
               
               
                 NM_000565 
                 IL6RA 
                 −0.67 
                 6365 
               
               
                 NM_002211 
                 ITGB1 
                 −0.65 
                 6366 
               
               
                 XM_049864 
                 CSF3 
                 −0.63 
                 6367 
               
               
                 XM_045933 
                 CAMKK2 
                 −0.63 
                 6368 
               
               
                 NM_033357 
                 CASP8 
                 −0.55 
                 6369 
               
               
                 XM_008704 
                 DNAM-1 
                 −0.52 
                 6370 
               
               
                 NM_030751 
                 TCF8 
                 −0.5 
                 6371 
               
               
                   
               
             
          
         
       
     
         [0205]     It is for example possible to take advantage of these characteristic changes in the method of the present invention.  
         [0206]     In the appended sequence listing, which is part of the present invention, the gene bank accession numbers (access via internet via http://www.ncbi.nlm.nih.gov/) indicated in tables 12 and 13 of the individual sequences are each allocated to one sequence ID. (SEQ. ID No.: 6243 to SEQ. ID No. 6372). The following sequence listing is part of the present invention.  
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