bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of PCR in Diagnosis and Prognosis of Visceral Leishmaniasis in Patients Coinfected with Human Immunodeficiency Virus Type 1
Authors: Pizzuto Massimo , Piazza Manuela , Senese Daniela , Scalamogna Chiara , Calattini Sara , Corsico Laura , Persico Tiziana , Adriani Beatrice , Magni Carlo , Guaraldi Giovanni , Gaiera Giovanni , Ludovisi Alessandra , Gramiccia Marina , Galli Massimo , Moroni Mauro , Corbellino Mario , Antinori Spinello 
Published: July 2002

ABSTRACT

A group of 76 consecutive human immunodeficiency virus (HIV)-positive patients with fever of unknown origin (
n
= 52) or fever associated with pulmonary diseases was evaluated in order to assess the usefulness of PCR with peripheral blood in the diagnosis and follow-up of visceral leishmaniasis. We identified 10 cases of visceral leishmaniasis among the 52 patients with fever of unknown origin. At the time of diagnosis, all were parasitemic by PCR with peripheral blood. During follow-up, a progressive decline in parasitemia was observed under therapy, and all patients became PCR negative after a median of 5 weeks (range, 6 to 21 weeks). However, in eight of nine patients monitored for a median period of 88 weeks (range, 33 to 110 weeks), visceral leishmaniasis relapsed, with positive results by PCR with peripheral blood reappearing 1 to 2 weeks before the clinical onset of disease. Eight
Leishmania infantum
and two
Leishmania donovani
infections were identified by PCR-restriction fragment length polymorphism analysis. PCR with peripheral blood is a reliable method for diagnosis of visceral leishmaniasis in HIV-infected patients. During follow-up, it substantially reduces the need for traditional invasive tests to assess parasitological response, while a positive PCR result is predictive of clinical relapse.
Methicillin-Resistant, Quinupristin-Dalfopristin-Resistant
Staphylococcus aureus
with Reduced Sensitivity to Glycopeptides
Authors: Werner G. , Cuny C. , Schmitz F.-J. , Witte W. 
Published: July 2002

ABSTRACT

Of 3,052
Staphylococcus aureus
strains collected by the European SENTRY surveillance study, 35 were found to be nonsusceptible to quinupristin-dalfopristin (MIC of ≥2 mg/liter). These isolates originated from four hospitals in France and one in Spain. In isolates from two Parisian hospitals exhibiting the same
Sma
I macrorestriction pattern, streptogramin resistance was based on
vatA
and
vgbA
. One isolate from a hospital in Lyon and 22 from a hospital in Lille were of the
vatB vgaB
streptogramin A resistance genotype and possessed
ermA
and/or
ermC
. As deduced from the loss of either streptogramin A or streptogramin B resistance determinants in particular isolates, resistance to quinupristin-dalfopristin requires mechanisms conferring resistance to both compounds. The
Sma
I macrorestriction patterns of strains from hospitals in Lille and Lyon were different; however, similarity analysis suggested a relatedness of 20 methicillin-resistant
S. aureus
strains from the Lille hospital, a finding confirmed by PCR typing based on three different genomic polymorphisms. These groups of isolates were found to be hetero-glycopeptide-intermediate susceptible
S. aureus
. Information about the failure of glycopeptide chemotherapy has not been available.
Identification of a
p28
Gene in
Ehrlichia ewingii
: Evaluation of Gene for Use as a Target for a Species-Specific PCR Diagnostic Assay
Authors: Gusa Asiya A. , Buller Richard S. , Storch Gregory A. , Huycke Mark M. , Machado Linda J. , Slater Leonard N. , Stockham Steven L. , Massung Robert F. 
Published: July 2002

ABSTRACT

PCR was used to amplify a 537-bp region of an
Ehrlichia ewingii
gene encoding a homologue of the 28-kDa major antigenic protein (P28) of
Ehrlichia chaffeensis
. The
E. ewingii p28
gene homologue was amplified from DNA extracted from whole blood obtained from four humans and one canine with confirmed cases of infection. Sequencing of the PCR products (505 bp) revealed a partial gene with homology to outer membrane protein genes from
Ehrlichia
and
Cowdria
spp.:
p30
of
Ehrlichia canis
(≤71.3%),
p28
of
E. chaffeensis
(≤68.3%), and
map1
of
Cowdria ruminantium
(67.3%). The peptide sequence of the
E. ewingii
partial gene product was deduced (168 amino acids) and the antigenicity profile was analyzed, revealing a hydrophilic protein with ≤69.1% identity to P28 of
E. chaffeensis
, ≤67.3% identity to P30 of
E. canis
, and ≤63.1% identity to MAP1 of
C. ruminantium
. Primers were selected from the
E. ewingii p28
sequence and used to develop a species-specific PCR diagnostic assay. The
p28
PCR assay amplified the expected 215-bp product from DNA that was extracted from EDTA-treated blood from each of the confirmed
E. ewingii
infections that were available. The assay did not produce PCR products with DNA extracted from
E. chaffeensis-
,
E. canis-
, or
E. phagocytophila
-infected samples, confirming the specificity of the
p28
assay for
E. ewingii
. The sensitivity of the
E. ewingii
-specific PCR assay was evaluated and determined to detect as few as 38 copies of the
p28
gene.
In Vitro Activity of Posaconazole against Clinical Isolates of Dermatophytes
Authors: Barchiesi Francesco , Arzeni Daniela , Camiletti Valeria , Simonetti Oriana , Cellini Andreina , Offidani Anna M. , Scalise Giorgio 
Published: July 2002

ABSTRACT

A broth macrodilution method following the recommendations established by the National Committee for Clinical Laboratory Standards was used to compare the in vitro activity of posaconazole (PCZ) with that of itraconazole (ITC) against 30 clinical isolates of dermatophytes belonging to six different species. In terms of MICs, PCZ showed an activity equal to that of ITC. MICs of PCZ at which 50% (MIC
50
) and 90% (MIC
90
) of the isolates were inhibited were 0.5 and >4.0 μg/ml, respectively. The MIC
50
and MIC
90
of ITC were 1.0 and >4.0 μg/ml, respectively. However, PCZ showed a more potent fungicidal activity than that of ITC against isolates belonging to the genus
Microsporum
(
P
= 0.03). PCZ merits further investigation as a potentially useful agent for treatment of dermatophytosis.
Kinetics of Dengue Virus-Specific Serum Immunoglobulin Classes and Subclasses Correlate with Clinical Outcome of Infection
Authors: Koraka P. , Suharti C. , Setiati T. E. , Mairuhu A. T. A. , Van Gorp E. , Hack C. E. , Juffrie M. , Sutaryo J. , Van Der Meer G. M. , Groen J. , Osterhaus A. D. M. E. 
Published: July 2002

ABSTRACTThe kinetics of dengue virus (DEN)-specific serum immunoglobulin classes (immunoglobulin M [IgM] and IgA) and subclasses (IgG1 to IgG4) were studied in patients suffering from dengue fever (DF), dengue hemorrhagic fever (DHF), and dengue shock syndrome (DSS). Serum samples from non-DEN febrile patients were included as controls. IgM, IgG1, and IgG3 serum antibodies were the predominant immunoglobulins throughout the course of illness in all three patient groups. In contrast, IgA antibodies were significantly higher in the acute phase in DSS patients compared to those in DF patients (P< 0.05). The levels of IgG1 differed significantly between patients with DF and those with DHF and DSS (P< 0.05). A significant difference was also found in IgG3 levels between DF patients and DHF patients (P< 0.05) but not between DF patients and DSS patients. Finally, levels of IgG4 antibodies differed significantly between DF patients and DSS patients (P< 0.05). Collectively, these data show that increased levels of DEN-specific IgA, IgG1, and IgG4 serum antibodies are risk markers for the development of DHF and DSS and that their measurement may provide valuable guidance for early therapeutic intervention.
Laboratory Diagnosis of Common Herpesvirus Infections of the Central Nervous System by a Multiplex PCR Assay
Authors: Markoulatos Panayotis , Georgopoulou Amalia , Siafakas Nikolaos , Plakokefalos Elias , Tzanakaki Georgina , Kourea-Kremastinou Jenny 
Published: July 2002

ABSTRACT
A sensitive multiplex PCR assay for single-tube amplification that detects simultaneous herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), varicella-zoster virus (VZV), human cytomegalovirus (CMV), and Epstein-Barr virus (EBV) is reported with particular emphasis on how the method was optimized and carried out and its sensitivity was compared to previously described assays. The assay has been used on a limited number of clinical samples and must be thoroughly evaluated in the clinical context. A total of 86 cerebrospinal fluid (CSF) specimens from patients which had the clinical symptoms of encephalitis, meningitis or meningoencephalitis were included in this study. The sensitivity of the multiplex PCR was determined to be 0.01 and 0.03 50% tissue culture infective doses/the reciprocal of the highest dilution positive by PCR for HSV-1 and HSV-2 respectively, whereas for VZV, CMV and EBV, 14, 18, and 160 ag of genomic DNA were detected corresponding to 48, 66, and 840 genome copies respectively. Overall, 9 (10.3%) of the CSF samples tested were positive in the multiplex PCR. HSV-1 was detected in three patients (3.5%) with encephalitis, VZV was detected in four patients (4.6%) with meningitis, HSV-2 was detected in one neonate (1.16%), and CMV was also detected in one neonate (1.16%). None of the samples tested was positive for the EBV genome. None of the nine positive CSF samples presented herpesvirus coinfection in the central nervous system. Failure of DNA extraction or failure to remove any inhibitors of DNA amplification from CSF samples was avoided by the inclusion in the present multiplex PCR assay of α-tubulin primers. The present multiplex PCR assay detects simultaneously five different herpesviruses and sample suitability for PCR in a single amplification round of 40 cycles with an excellent sensitivity and can, therefore, provide an early, rapid, reliable noninvasive diagnostic tool allowing the application of antiviral therapy on the basis of a specific viral diagnosis. The results of this preliminary study should prompt a more exhaustive analysis of the clinical value of the present multiplex PCR assay.
Rapid Identification of Methicillin-Resistant
Staphylococcus aureus
from Positive Blood Cultures by Real-Time Fluorescence PCR
Authors: Tan Thean Yen , Corden Sally , Barnes Rosemary , Cookson Barry 
Published: July 2002

ABSTRACT

Methicillin-resistant
Staphylococcus aureus
septicemia is associated with significant morbidity and mortality and requires treatment with intravenous glycopeptides. For blood cultures positive for gram-positive cocci, 24 to 48 h is required for the detection of
S. aureus
bacteremia and the provision of antibiotic susceptibility testing results. We describe a molecular biology-based assay that requires 2 h from the time of initial positivity of blood cultures. The assay correctly detected 96% of the
S. aureus
isolates including all methicillin-resistant
S. aureus
isolates. Clinical data collected during the study suggest that 28% of patients with
S. aureus
bacteremia do not receive early and appropriate treatment and that 10% of patients may initially be receiving inappropriate glycopeptide treatment.
Risk ofMycobacterium tuberculosisTransmission in a Low-Incidence Country Due to Immigration from High-Incidence Areas
Authors: Lillebaek Troels , Andersen Åse B. , Bauer Jeanett , Dirksen Asger , Glismann Steffen , de Haas Petra , Kok-Jensen Axel 
Published: July 2002

ABSTRACTDoes immigration from a high-prevalence area contribute to an increased risk of tuberculosis in a low-incidence country? The tuberculosis incidence in Somalia is among the highest ever registered. Due to civil war and starvation, nearly half of all Somalis have been forced from their homes, causing significant migration to low-incidence countries. In Denmark, two-thirds of all tuberculosis patients are immigrants, half from Somalia. To determine the magnitude ofMycobacterium tuberculosistransmission between Somalis and Danes, we analyzed DNA fingerprint patterns of isolates collected in Denmark from 1992 to 1999, comprising >97% of all culture-positive patients (n= 3,320). Of these, 763 were Somalian immigrants, 55.2% of whom shared identical DNA fingerprint patterns; 74.9% of these were most likely infected before their arrival in Denmark, 23.3% were most likely infected in Denmark by other Somalis, and 1.8% were most likely infected by Danes. In the same period, only 0.9% of all Danish tuberculosis patients were most likely infected by Somalis. The Somalian immigrants in Denmark could be distributed into 35 different clusters with possible active transmission, of which 18 were retrieved among Somalis in the Netherlands. This indicated the existence of some internationally predominant Somalian strains causing clustering less likely to represent recent transmission. In conclusion,M. tuberculosistransmission among Somalis in Denmark is limited, and transmission between Somalis and Danes is nearly nonexistent. The higher transmission rates between nationalities found in the Netherlands do not apply to the situation in Denmark and not necessarily elsewhere, since many different factors may influence the magnitude of active transmission.
Typing of
Candida albicans
Isolates by Sequence Analysis of the Cytochrome
b
Gene and Differentiation from
Candida stellatoidea
Authors: Biswas Swarajit Kumar , Yokoyama Koji , Wang Li , Nishimura Kazuko , Miyaji Makoto 
Published: July 2002

ABSTRACT

Including type strains, mitochondrial cytochrome
b
genes of 32 strains of
Candida albicans
and 6 strains of
Candida stellatoidea
, presently treated as a synonym for
C. albicans
, were partially sequenced. Analysis of 396-bp nucleotide sequences of the strains under investigation divided
C. albicans
isolates into three types: type I, type II, and type III; however, strains of
C. stellatoidea
represented distinct type IV isolates. Deduced amino acid sequences of type I, type II, and type III were identical and differed from that of type IV by one amino acid. Genotypes (rDNA type) of the test strains were also checked. Cytochrome
b
typing did not correlate with genotyping, and different genotypes occurred for one cytochrome
b
type. This study shows that cytochrome
b
gene sequences are useful for analyzing the genetic relatedness of
C. albicans
isolates and effective for differentiating
C. stellatoidea
from
C. albicans
.
Genotyping
Encephalitozoon hellem
Isolates by Analysis of the Polar Tube Protein Gene
Authors: Xiao Lihua , Li Lixia , Moura Hercules , Sulaiman Irshad , Lal Altaf A. , Gatti Simonetta , Scaglia Massimo , Didier Elizabeth S. , Visvesvara Govinda S. 
Published: July 2002

ABSTRACT

To develop an alternative genotyping tool, the genetic diversity of
Encephalitozoon hellem
was examined at the polar tube protein (PTP) locus. Nucleotide sequence analysis of the PTP gene divided 24
E. hellem
isolates into four genotypes, compared to two genotypes identified by analysis of the internal transcribed spacer of the rRNA gene. The four PTP genotypes differed from each other by the copy number of the 60-bp central repeat as well as by point mutations. A simple PCR test was developed to differentiate
E. hellem
genotypes based on the difference in the size of PTP PCR products, which should facilitate the genotyping of
E. hellem
in clinical samples.
Clinical Isolates of Non-O157 Shiga Toxin-Producing
Escherichia coli
: Serotypes, Virulence Characteristics, and Molecular Profiles of Strains of the Same Serotype
Authors: Eklund Marjut , Scheutz Flemming , Siitonen Anja 
Published: July 2002

ABSTRACT

All human Shiga toxin-producing
Escherichia coli
(STEC) non-O157 strains (
n
= 56) isolated in Finland from 1990 to August 2000 were characterized for the O:H serotype,
stx
1
and
stx
2
genes, production of enterohemolysin, and sensitivity to 12 antimicrobial agents. Strains of the same serotype were genotyped by pulsed-field gel electrophoresis (PFGE) after
Xba
I restriction of total DNA. The 56 non-O157 isolates belonged to 29 serotypes. Two of the serotypes (O102:H7 and OX181:H49) have not previously been described as being associated with STEC infections in humans or isolated from animals. Thirty-four strains (61%) within seven serotypes (O103:H2 [14 isolates], O26:H11 [6 isolates], O145:H28 [4 isolates], O145:HNM [3 isolates], O15:HNM [3 isolates], OX174:H21 [2 isolates], and O Rough:HNM [2 isolates]) were represented by more than one isolate. Of these strains, O103:H2 isolates were divided into seven, O26:H11 isolates were divided into four, and the rest within a serotype were divided into two genotypes in PFGE. In PCR, 31 (55%) of the 56 strains were positive for the
stx
2
gene only and 24 strains (43%) were positive for
stx
1
only. One strain (O43:H2) carried both
stx
1
and
stx
2
. Forty-two strains (75%) produced enterohemolysin, and 39 strains (70%) possessed the
eae
gene. Of the latter 39 strains, 36 (92%) were enterohemolytic, whereas only 6 (35%) of the 17 isolates lacking the
eae
gene were enterohemolytic (
P
< 0.001). The majority of the strains (44 strains, 79%) were sensitive to all 12 antimicrobials tested. Of the 56 strains, 20 (36%) were associated with small family outbreaks in nine families and 14 (25%) were associated with recent travel abroad.
Detection of Ebola Viral Antigen by Enzyme-Linked Immunosorbent Assay Using a Novel Monoclonal Antibody to Nucleoprotein
Authors: Niikura Masahiro , Ikegami Tetsuro , Saijo Masayuki , Kurane Ichiro , Miranda Mary E. , Morikawa Shigeru 
Published: July 2002

ABSTRACT
With the increase in international traffic, the risk of introducing rare but severe infectious diseases like Ebola hemorrhagic fever is increasing all over the world. However, the system for the diagnosis of Ebola virus infection is available in a limited number of countries. In the present study, we developed an Ebola virus antigen-detection enzyme-linked immunosorbent assay (ELISA) system using a novel monoclonal antibody (MAb) to the nucleoprotein (NP). This antibody recognized an epitope defined by a 26-amino-acid stretch near the C terminus of NP. In a sandwich ELISA system with the MAb, as little as 30 ng of purified recombinant NP (rNP) was detected. Although this MAb was prepared by immunization with rNP of subtype Zaire, it also reacted to the corresponding region of NP derived from the Reston and Sudan subtypes. These results suggest that our ELISA system should work with three of four Ebola subtypes. Furthermore, our ELISA system detected the NP in subtype Reston-infected monkey specimens, while the background level in noninfected specimens was very low, suggesting the usefulness of the ELISA for laboratory diagnosis with clinical specimens.
Evaluation of Recombinant
Leptospira
Antigen-Based Enzyme-Linked Immunosorbent Assays for the Serodiagnosis of Leptospirosis
Authors: Flannery Brendan , Costa Dirceu , Carvalho Fernanda Pinheiro , Guerreiro Hygia , Matsunaga James , Da Silva Emilson Domingos , Ferreira Antonio Gomes Pinto , Riley Lee W. , Reis Mitermayer G. , Haake David A. , Ko Albert I. 
Published: July 2002

ABSTRACT

There is an urgent need for development of new serodiagnostic strategies for leptospirosis, an emerging zoonosis with worldwide distribution. We have evaluated the diagnostic utility of five recombinant antigens in enzyme-linked immunosorbent assays (ELISAs) for serodiagnosis of leptospirosis. Sera from 50 healthy residents of a high-incidence region were used to determine cutoff values for 96% specificity. In paired sera from 50 cases of leptospirosis confirmed by the microscopic agglutination test, immunoglobulin G (IgG) but not IgM reacted with the recombinant leptospiral proteins. The recombinant LipL32 IgG ELISA had the highest sensitivities in the acute (56%) and convalescent (94%) phases of leptospirosis. ELISAs based on recombinant OmpL1, LipL41, and Hsp58 had sensitivities of 16, 24, and 18% during the acute phase and 72, 44, and 32% during convalescence, respectively. Compared to sera from healthy individuals, patient sera did not react significantly with recombinant LipL36 (
P
> 0.05). Recombinant LipL32 IgG ELISA demonstrated 95% specificity among 100 healthy individuals, and specificities ranging from 90 to 97% among 30 dengue patients, 30 hepatitis patients, and 16 patients with diseases initially thought to be leptospirosis. Among 39 Venereal Disease Research Laboratory test-positive individuals and 30 Lyme disease patients, 13 and 23% of sera, respectively, reacted positively with the rLipL32 antigen. These findings indicate that rLipL32 may be an useful antigen for the serodiagnosis of leptospirosis.
Hemorrhagic Fever with Renal Syndrome: Diagnostic Problems with a Known Disease
Authors: Wichmann Dominic , Slenczka Werner , Alter Peter , Boehm Stephan , Feldmann Heinz 
Published: July 2002

ABSTRACT
Hemorrhagic fever with renal syndrome (HFRS), caused by different hantaviruses, is a distinct clinical syndrome endemic in several parts of Asia and Europe. However, the clinical picture can sometimes be indistinguishable from that of other infectious or noninfectious diseases. In this report we describe a clinical case, which is a rare occurrence but is a prime example of the difficulties in the diagnosis of HFRS in areas with a low prevalence of the disease.
Phenotypic Variation ofHelicobacter pyloriIsolates from Geographically Distinct Regions Detected by Lectin Typing
Authors: Hynes Sean O. , Broutet Nathalie , Eurohepygast Study Group  , Wadström Torkel , Mikelsaar Marika , O’Toole Paul W. , Telford John , Engstrand Lars , Kamiya Shigeru , Mentis Andreas F. , Moran Anthony P. 
Published: July 2002

ABSTRACTA total of 309Helicobacter pyloriisolates from 18 different countries were analyzed with a previously developed lectin typing system. The system was developed by using a proteolytic pretreatment to enhance the carbohydrate fraction of the sample. Four lectins fromUlex europaeus,Lotus tetragonolobus,Erythrina cristigali, andTriticum vulgariswere used to type the strains. The lectins were chosen for their specificities for sugars commonly encountered in the lipopolysaccharide ofH. pylori. The isolates were received from their parent institutions as pellets of biomass and were typed at one of three centers (in Ireland, Sweden, and Estonia). All 16 possible lectin reaction patterns were observed in the study, with the isolates with the predominant pattern exhibiting reactions with all the lectins in the panel. For European patients suffering from gastritis, an association was noted between lectin reaction pattern MH4 and atrophic chronic gastritis; isolates with lectin reaction pattern MH4 were isolated from patients with atrophic chronic gastritis, whereas isolates with this pattern were not isolated from patients with chronic gastritis (P= 0.0006). In addition, statistically significant relationships were noted between the lectin reaction pattern and the associated pathology of isolates from the Swedish population. Isolates with patterns MH13 and MH16, which had low lectin reactivities, correlated with nonulcer disease (P= 0.0025 andP= 0.0002, respectively), and all four isolates from adenocarcinoma patients were characterized as possessing reaction pattern MH16. In contrast, isolates with lectin reaction patterns MH1 and MH10, which had high lectin reactivities, were associated with ulcer disease (P= 0.046 andP= 0.0022, respectively).
Molecular Typing of
Treponema pallidum
in South Africa: Cross-Sectional Studies
Authors: Pillay A. , Liu H. , Ebrahim S. , Chen C. Y. , Lai W. , Fehler G. , Ballard R. C. , Steiner B. , Sturm A. W. , Morse S. A. 
Published: July 2002

ABSTRACT

We evaluated a molecular subtyping system for
Treponema pallidum
for its ability to differentiate between strains obtained from male patients with primary syphilis in South Africa. Of 201
T
.
pallidum
-positive specimens, 161 were typeable, revealing 35 subtypes. The unique subtypes identified in Durban, Cape Town, and Carletonville and the total number of subtypes suggested that the strain population was very diverse and varied geographically.