Abstract:
The present invention seeks to provide an express-control system for analyzing the compatibility of blood with other substances. The present invention relates to a system, method and apparatus for use in blood testing and more specifically, food intolerance testing through analysis of blood using luminescence. The present invention utilizes a blood testing system and apparatus, which includes a displacement assembly, a blood divider assembly, a reagent divider assembly, an electro-optical multiplier assembly and a central controller. A sample of blood is added to a corresponding one of a plurality of canisters containing a test substance. Thereafter a luminescent reagent is added and the plurality of canisters are moved in the blood testing apparatus until the canisters begin to luminese. The amount of luminescence is measured and analyzed an indicator of compatibility of the blood with the test substance and more specifically as an indicator of the presence and activity of neutrophil in the blood and more specifically the granulacy of the blood.

Description:
FIELD OF THE INVENTION 
     The present invention relates to a system, apparatus and method for use in blood testing and more specifically, food intolerance testing through analysis of blood using luminescence. The present invention also relates to a system, apparatus and method for successively testing blood to determine the compatibility of blood to different test substances using the luminescence as an indicator of the amount and/or activity of neutrophil in the blood. 
     BACKGROUND OF THE INVENTION 
     The immune system is an intricate collection of organs, tissues, cells and soluble factors that allow individuals to defend against harmful agents such as viruses, bacteria, fungi, parasitic organisms, and tumor cells. The immune system is a recognition system that distinguishes the body&#39;s own molecules from foreign molecules. When the immune system detects a foreign substance, called an antigen, it responds with a proliferation of cells that either attack the invader directly (the innate immune response) or produce specific defensive proteins called antibodies (the adaptive immune response). In contrast to the innate immune defences, which are always ready to fight a variety of infections, the adaptive immune response must be primed by the presence of an antigen, and the defensive cells and antibodies produced against the antigen are ineffective against any other foreign substance. Typically, the innate immune response has a rapid reaction time, whereas the adaptive immune response has a slow initiation and increases thereafter. 
     Allergies are hypersensitivities of the body&#39;s defence system to certain environmental antigens. Allergic reactions are typically very rapid and show extraordinary sensitivity to minute amounts of antigen. When antibodies that participate in allergenic reactions bind to antigens, a process called degranulation occurs in which the masked cells release a flood of histamine and other inflammatory mediators which cause some of the best recognized systems of allergy, namely sneezing, nasal irritation, itchiness of the skin and tearing of the eyes. 
     The presence of biologically active agents including antigens in a patient&#39;s body fluid, especially blood, has been determined using various techniques. One such technique entails the analysis of either bioluminescence or chemiluminescence for detecting the presence of a variety of luminescent analytes. For example, U.S. Pat. No. 5,445,794 discloses a luminescence measuring system comprising a luminometer designed for making bioluminescence and chemiluminescence measurements. The disclosed luminometer comprises a chamber into which a single test tube is received. The test tube once received in the chamber is held stationary while the chamber revolves around the test tube from the admission phase to the measurement phase and finally to the discharge phase. It is in the measurement phase that the reagent LUMIT is added to the sample and luminescence is measured. The disadvantage of the technique and apparatus disclosed in U.S. Pat. No. 5,445,794 is that it allows for only a single sample to move through the testing process from admission to discharge and as such, results in decreased efficiency and increased time in the overall testing process. 
     U.S. Pat. No. 6,335,166 discloses an automated analyser capable of performing multiple diagnostic assays simultaneously. The disclosed analyser includes a computer controller which runs analyser-controlling and assay-scheduling software to coordinate operation of the stations of the analyser and movement of each reaction receptacle through the analyser. The specimen pipette assembly of the disclosed analyser is coupled to a syringe pump which engages the specimen tubes carried on the specimen ring and which also engages pipette tips carried on a pipette wheel near the back portion of the specimen ring. The containers of the target capture reagent are carried on an inner rotable assembly constructed and arranged to selectively agitate the containers or present the containers for access by the probe of an automatic robotic pipette system. The reaction mixtures are prepared by the pipette system within each reaction receptacle. The contents of the reaction receptacle is subjected to magnetic separation wash procedures in the magnetic separation wash stations of the apparatus. The disclosed apparatus also contains a luminometer for detecting and/or quantifying the amount of light emitted by the contents of the reaction receptacle. The disclosed apparatus of U.S. Pat. No. 6,335,166 requires treatment of the contents of the reaction receptacle by magnetic separation wash procedures. Furthermore, the disclosed apparatus consists of several interconnected ring assemblies resulting in an overly complex and intricate apparatus. 
     Likewise, U.S. Pat. No. 3,617,222 discloses a complex apparatus and method for testing and classifying materials, which contain agglutinates. The method taught by U.S. Pat. No. 3,617,222 enables agglutinations to be detected either by nephelometry or by opacimetry and does not disclose a method for luminescence detection. The disclosed apparatus consists of an agitator comprising a turn-table subjected to motion around a circle in which the reaction mixture is introduced into the test vessel via two syringes. The first syringe is used to take small amounts of the sample, the second used to take into a small bottle a certain amount of liquid containing reagents, dilutant and flush water. A mass of liquid taken from the first and second syringes is directed into a probe which contains the sample of the reagent and dilutant, the contents of the probe being pushed into the reaction cups which are then subject to agitation. The apparatus taught in U.S. Pat. No. 3,617,222 discloses a two stage mixing procedure comprising a partial mixing in the probe and then a more complex mixing by agitation. Such a procedure disadvantageously results in an intricate preparation of the reaction mixture prior to the actual testing phase, thereby disadvantageously increasing the total time in the overall testing process. 
     U.S. Pat. No. 5,422,075 teaches a chemical luminescence-detection apparatus in which the chemical luminescence generated in a photometric cell is detected by an optical detector. The reagent used in the luminescent reaction is luminol. U.S. Pat. No. 5,422,075 also requires complex preparation of the sample prior to the testing phase and the luminescent reaction. The requisite preparation includes subjecting the reaction solution to several agitations and washes. Such a complex preparation of the sample disadvantageously increases the time required in the overall testing process. 
     SUMMARY OF THE INVENTION 
     Accordingly, it is an advantage of the present invention to provide an express-control system for analysing the compatibility of blood with other substances. 
     It is also an advantage of the present invention to provide an efficient system for blood testing using luminescence, which does not require complex preparation of the sample of blood prior to testing. 
     It is a further advantage of the present invention to provide a blood analyser which is able to test several substances concurrently or individually. 
     It is also an advantage of the present invention to provide a blood analyser, which is simple in design and easily disassembled for maintenance by the operator. 
     In one aspect, the present invention seeks to provide an efficient system for blood testing which is able to more efficiently and quickly test the compatibility of blood with various other substances without requiring complex preparation of the sample of blood prior to testing. Thereby, the present invention advantageously decreases the time required in the overall testing process by eliminating the expensive and time consuming steps associated with the complex preparation of the reaction mixture. The apparatus of the present invention is easy to use and advantageously facilitates easy maintenance by the user. 
     In another aspect, the present invention resides in a blood testing system for testing the compatibility of blood with different test substances, said system comprising: a displacement assembly comprising a plurality of canister holders, each canister holder adaptable to hold a canister, each canister initially containing a corresponding one of the test substances, said displacement assembly moving each of the plurality of canisters to a sampling stage, a reagent adding stage and an optical detector stage; a blood divider assembly at the sampling stage, said blood divider assembly operable to add a predetermined amount of the blood to each of the plurality of canisters containing the corresponding test substance as the canisters are moved to the sampling stage; a reagent divider assembly at the reagent adding stage, said reagent divider assembly operable to add a predetermined amount of luminescent reagent to each of the plurality of canisters as the canisters are moved to the reagent adding stage; an electro-optical multiplier assembly at the optical detector stage, the electro-optical multiplier assembly operable to measure luminescence of each of the plurality of canisters at a predetermined time period after the luminescent reagent has been added to the corresponding canister; and a central controller operable to control the functions of the drum assembly, the blood divider assembly, the reagent divider assembly and the electro-optical multiplier assembly and to process data received therefrom and assess the compatibility of the blood to each of the test substances. 
     In yet another aspect, the present invention resides in a blood analyser comprising: a displacement instrument, the displacement instrument comprising an admission site, a sample adding site, a reagent adding site and a discharge site; a plurality of canisters holders adaptable to hold a plurality of canisters, each canister initially containing a different test substance, wherein the plurality of canister holders are attached to the displacement instrument for movement from the admission site to the discharge site; a sample divider for supplying a predetermined amount of a blood sample to each of the plurality of canisters at the sample adding site; a reagent divider for supplying a predetermined amount of luminescent reagent to each of the plurality of canisters at the reagent adding site; a detection instrument for detecting luminescence after the luminescent reagent is added to each of the plurality of canisters; a converting instrument for converting the detected luminescence into a luminescence signal; and a central controller operable to control the functioning of the blood analyser and process data received from the blood analyser and assess the compatibility of the blood to each of the test substances. 
     In still another aspect, the present invention resides in a method for measuring the compatibility of blood with test substances using luminescence, the method comprising: adding a sample of blood to each of a plurality of canisters, each of the plurality of canisters containing a different test substance; adding a luminescent reagent to each of the plurality of canisters with the sample of blood and the test substance; subjecting the canisters to movement for a predetermined period of time until the canisters begin to luminescence; and detecting the amount of luminescence by a photometric measurement. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       Further aspects of the present invention will become apparent upon reading the following detailed description together with the accompanying drawings in which: 
         FIG. 1  is a top view of the blood testing system according to an embodiment of the present invention; 
         FIG. 2  is a bottom view of the blood testing system of  FIG. 1  according to an embodiment of the present invention 
         FIG. 3  is a side view of the blood testing system of  FIG. 1  showing the connectors in accordance with a preferred embodiment of the invention; 
         FIG. 4  is a flowchart diagram of a preferred method of operation of the blood testing system of  FIG. 1  according to an embodiment of the invention; and 
         FIG. 5  is an algorithm of a preferred mode of operation of the blood testing system of  FIG. 1  according to an embodiment of the invention. 
     
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
       FIG. 1  is a top view of the blood testing system  2  according to a preferred embodiment of the present invention. The blood testing system  2  includes a displacement assembly  4 , a blood divider assembly  6 , a reagent divider assembly  7 , an electro-optical multiplier assembly  8  and a central controller  10 . 
     The displacement assembly  4  in one embodiment comprises a drum  12 . The drum  12  includes a hollow cylindrical body with a circular base (not shown), a wall of constant circular cross-section (not shown) and a lid  14 . The interior of drum  12  contains a plurality of canister holders  18 . The canister holders  18  are adaptable to hold a plurality of canisters  20  each containing a test substance. In a preferred embodiment, the test substance in each of the plurality of canisters  20  can be one of a variety of different antigens. The lid  14  contains an opening  22  through which each of the plurality of canisters  20  can be inserted into the canister holders  18 . Preferably the canisters  20  are test tubes with opaque walls. The displacement assembly  4  is operable to move each of the plurality of canisters  20  to an admission site  1 - 1 , a sample adding site  1 - 2  in the sampling stage, a reagent adding site  1 - 3  in the reagent adding stage and a discharge site  1 - 4  (shown in  FIG. 2 ) after an optical detection stage. 
     The plurality of the canisters  20  once inserted into the displacement assembly  4 , at the admission site  1 - 1 , are moved to the sampling site  1 - 2  where a sample of blood is added to each of the plurality of canisters  20  during the sampling stage. 
     At the sampling site  1 - 2 , the blood divider assembly  6  includes a blood container  23 , preferably a syringe, and canula  24  connectable to the blood container  23 . The blood container  23  contains therein a sample of blood drawn from a subject and is operable to add a predetermined amount of the blood to a corresponding one of the plurality of canisters  20  containing the test substance. The blood divider assembly  6  further comprises a piston  34  coupled to the blood container  23  and canula  24 , a piston connector  36  connecting the piston  34  to the central controller  10  and a blood divider sensor  32  operatively positioned adjacent the blood divider assembly  6 , and coupled to the central controller  10 . The blood divider sensor  32  is operable to detect the amount of blood dropped into each of the plurality of canisters  20 , generate a blood sampling signal indicative thereof and send the blood sampling signal to the central controller  10 . Movement of the piston  34  by the central controller  10  affects the release of a predetermined amount of blood from the blood container  23  and canula  24  into the corresponding canister of the plurality of canisters  20  containing the test substance. The dimension of the canula  23  is correlated to the amount the piston  34  must move in order to obtain the correct predetermined amount of blood in each of the plurality of canisters  20 . 
     The plurality of canisters  20  are then moved to the reagent adding site  1 - 3  where a sample of reagent is added during the reagent adding stage. 
     At the reagent adding site, the reagent divider assembly  7  is operable to add a predetermined amount of the luminescent reagent to each of the plurality of canisters  20  containing both the test substance and the predetermined amount of blood. The reagent divider assembly  7  includes a reagent container  26  and an electro-pneumatic block assembly  40 . The luminescent reagent, preferably Luminol, is held within the reagent container  26  while the electro-pneumatic block assembly  40  regulates the amount of reagent released from the reagent container  26  into each of the plurality of canisters  20 . 
     After the amount of reagent is added to each of the plurality of canisters  20 , the canisters  20  are moved to the optical detection stage. 
     At the optical detector stage, the electro-optical multiplier assembly  8  is electronically coupled to the drum  12  of the displacement assembly  4  and is operable to detect and measure photons or luminescence emanating from the plurality of canisters  20 . The electro-optical multiplier assembly  8  comprises an electro-optical multiplier  42  and at least one photosensitive cathode  44 . The electro-optical multiplier  42  transforms the luminescence detected by the at least one photosensitive cathode  44  into an electrical signal and transmits the electrical signal to the central controller  10 . 
     The central controller  10  is operatively coupled to each of the displacement assembly  4 , the blood divider assembly  6 , the reagent divider assembly  7  and the electro-optical multiplier assembly  8 . The central controller  10  is operable to control the functions of each of the aforementioned assemblies and is also operable to process data received therefrom and assess the compatibility of the blood to each of the test substances. The central controller  10  includes input ports (not shown), output ports (not shown), circuitry (not shown), a keypad  46  and an LCD display  48 . 
     Referring now to  FIG. 2  a bottom view of the blood testing system  2  in accordance with an embodiment of the present invention is shown. A base  49  of the drum  12  contains a canister discharge opening  50  and a discharge mechanism  52  at the discharge site  1 - 4  operable to discharge the canisters from the displacement assembly after use. The discharge mechanism  52  is preferably a flexible hose adaptable to be directed to a waste container (not shown). The displacement assembly  4  further comprises a drum rotator  56  and a secondary drum rotator  62 . 
     The drum rotator  56  is operable to rotate the drum from the admission site  1 - 1  through the sampling stage, the reagent adding stage and the optical detector stage to the discharge site  1 - 4 . The drum rotor  56  is mechanically coupled to a drum flywheel  57 . The drum rotor  56  is operable to rotate the drum flywheel  57  and thereby also rotate the drum  12 . The drum rotor  56  also includes a first optical sensor  58  positioned adjacent to the drum  12  and which is operable to generate a drum positioning signal indicative of the rotation and the annular position of the drum  12  during rotation. The first optical sensor  58  is electronically coupled to the central controller  10  and sends the drum position signal to the central controller  10 . The drum rotor  56  also comprises a second optical sensor  60  operatively positioned adjacent to the canister holders  18 . The second optical sensor  60  is operable to generate a canister positioning signal indicative of the positioning of the canisters  20  within the drum  12  and to send the canister positioning signal to the central controller  10 . The central controller  10  receives the drum positioning signal and the canister positioning signal and sends control signals to the displacement assembly  4  to control the displacement assembly  4 . 
     The secondary drum rotator  62  is also coupled to drum  12 . The secondary drum rotator  62  is operable to control friction of the drum  12  during rotation, and is also operable to prevent jamming of the plurality of canisters  20  contained therein. The secondary drum rotator  62  is also electronically coupled to the central controller  10 , and is operable to receive the control signals therefrom. In response to the control signals received from the central controller  10 , the secondary drum rotor  62  is able to adjust the rotation of the drum  12 . 
       FIG. 3  shows a side view of the blood testing system  2  of  FIG. 1  in accordance with a preferred embodiment of the present invention. The reagent container  26  includes a top portion  66  and a bottom portion  68 . The top portion  66  is removable to allow for the reagent container  26  to be filled with the luminescent reagent. 
     The reagent divider assembly  7  further comprises an electro-pneumatic block assembly  40 , which is operable to regulate the amount of reagent released from the reagent container  26 . The electro-pneumatic block assembly  40  includes an air compressor  74 , a hose  72 , a valve  78  and an electro-pneumatic block connector  86 . The top portion  66  of the reagent container  26  contains a fitting  70  to which the hose  72  is attached. The hose  72  is also attached to the air compressor  74  to allow air to pass from the air compressor  74  into the reagent container  26 . The bottom portion  68  of the reagent container  26  also contains a fitting  76  to which the valve  78  is attached on an inlet side  86  thereof. A disconnection safety cup  84 , which is operable to prevent leakage from the electro-pneumatic valve  78  when not in use, is attached to an outlet side  82  of the valve  78 . The outlet side  82  of the valve  78  is situated overhead of the plurality of canisters  20 . The valve  78  is connected to the central controller  10  by the electro-pneumatic block connector  86 . The valve  78  is operable to allow a predetermined amount of reagent to pass from the reagent container  26  into a corresponding one of the plurality of canisters  20 . 
     A panel of the central controller is also shown including a power on/off switch  90 , a USB PC connection Type B  92 , a COM PC connection port  94 , and a Blood-dozing system control connection port  96 . 
     Referring now to  FIG. 4 , a flowchart diagram of a preferred method of operation of the blood testing system  2  of  FIG. 1 , in accordance with an embodiment, is shown. 
     In a pre-testing stage a sample of blood is drawn from the subject into a syringe  28  and the needle  100 . An anticoagulant, preferably heparin is also added to the sample of blood in the syringe  28 . In a preferred embodiment the blood undergoes no further treatments, or washes other than the addition of the anticoagulant. The needle  100  of the syringe  28  is then replaced by the canula  24 . The syringe  28  and canula  24  are then inserted into the blood divider assembly  6 . 
     During the sampling stage  102  each of the plurality of canisters  20  pre-filled with the test substance, are moved into place beneath the blood divider assembly  6 . A predetermined amount of blood, approximately two or three drops, is then added. During the next stage, the reagent adding stage  104 , each of the plurality of canisters  20  with the test substance and the sample of blood are moved into place beneath the reagent divider assembly  7 , where a predetermined amount of reagent, preferably Luminol, is added to the reaction mixture in each of the plurality of canisters  20 . By way of non-limiting example, Luminol is added in an amount between 0 to 100 ml. 
     Each of the plurality of canisters  20  containing the test substance, blood, and Luminol are then moved for a predetermined period of time after which the canisters begin to luminese. In a preferred embodiment, the period of time which the canisters are moved corresponds approximately to the time taken for the displacement assembly  4  to move each of the plurality of canisters  20  from the reagent adding stage  104  to the optical detector stage  106 . At the optical detector stage  106  the total luminescence is detected by the electro-optical multiplier assembly  8 , converted into an electrical signal (a luminescence signal) and sent to the central controller  10 . Thereafter each of the plurality of canisters  20  are successively ejected from the displacement assembly  4  into a waste receptacle (not shown). 
     The luminescence signal generated by the electro-optical multiplier  42 , once received in the central controller  10  is processed in a measurement stage  108 . During the measurement stage  108  the received information is analysed through a software program designed to measure the luminosity as an indicator of at least one of the presence and activity of neutrophil in the blood and more specifically the granulacy of the blood. While it has been disclosed that the presence and activity of neutrophil in the blood is measured, a skilled artisan could envision alternate embodiments that are possible utilising the present invention as for example, the presence of Eosinophil in the blood. 
     Although it has been described in the preferred embodiment that the sampling stage  102  and the reagent adding stage  104  occur separately, it is to be understood that this is but one embodiment of the invention. A skilled artisan would readily appreciate that the sampling stage  102  and reagent adding stage  104  could occur concurrently to achieve the objects of the present invention. 
     Referring now to  FIG. 5  an algorithm is shown for a preferred mode of operation of the blood testing system  2  of  FIG. 1  according to an embodiment of the invention. 
     The central controller  10  processes data received from the various assemblies of the system  2  to assess the compatibility of the blood to each of the test substances. The central controller  10  receives the luminescence signal for each of the plurality of canisters  20 , correlates the signal to the corresponding test substance and generates a compatibility output for each of the test substances based on the luminescence signal. 
     After the blood testing system starts  109  it performs five measurements  110  generating data which is then normalized to values between 0 and 1000 at operation  112 . Thereafter the root mean square deviation (RMS) for the normalized data is calculated according to the formula √[(Σx 2 )/n] at operation  114 . Corridors are built for the values RMS, 2*RMS and 3*RMS at operation  116 . Thereafter the remaining measurements are performed at operation  117 . If none of the sample points in the interval 1 . . . 10 seconds are outside 2*RMS at operation  118 , then the reading is “Neutral” at operation  120  and the system proceeds to the “Finish” operation  130 . If any of the sample points in the interval 1 . . . 10 seconds are outside 2*RMS at operation  122  then the reading is “Mild” at operation  124  and the system proceeds to the “Finish” operation  130 . If any of the sample points in the interval 1 . . . 10 seconds are outside 3*RMS at operation  126  then the reading is “Severe” at operation  128  and the system proceeds to the “Finish” operation  130 . The aforementioned measurements of “Neutral”, “Mild” and “Severe” relate to the level of compatibility of the blood to the specific antigen. 
     Accordingly, in one embodiment, the luminescence signal comprises at least 3 luminescence sample measurements for each of the plurality of canisters, and, the central controller generates the compatibility output for each of the test substances by determining a root mean square deviation of each of the at least 3 measurements and generating a neutral output for the test substance if none of the sample measurements are outside of two times the root mean square, generating a mild output for the test substance if at least one of the sample measurements are outside of two times the root mean square, and, generating a severe output for the test substance if at least one of the sample measurements is outside of three times the root mean square. 
     In another embodiment, the luminescence signal comprises at least 5 luminescence sample measurements for each test sample, and, the electro-optical multiplier acquires the at least 5 sample measurements for each of the plurality of canisters within 5 to 15 seconds. 
     In yet another embodiment, the central controller outputs one of the neutral output, the mild output and the severe output for each of the test substances indicating the compatibility of the blood to each of the test substances in the plurality of canisters. 
     It is to be understood that all the various features of the invention have been described with respect to one or another of the embodiments in the invention, and that the various features and embodiments of the invention may be combined or used in combination with other features and embodiments of the invention as described and illustrated herein. 
     Furthermore although this disclosure has been described and illustrated as containing preferred embodiments of the invention, it is to be understood that the invention is not restricted to these particular embodiments. Rather, the invention includes all embodiments, which are functional, electrical, electronical or mechanical equivalents of the specific embodiments and features that have been described herein. It is also to be understood that other types of testing could be achieved through utilizing the system, method and apparatus of the present invention, as for example blood compatibility with tissue cells from transplanted organs.