bluuebunny/crossref_metadata_2025_split huggingface dataset:
Leishmania major-Like Antigen for Specific and Sensitive Serodiagnosis of Human and Canine Visceral Leishmaniasis
Authors: Barbosa-de-Deus Rosângela , dos Mares-Guia Marcos Luíz , Nunes Adriane Zacarias , Costa Kátia Morais , Junqueira Roberto Gonçalves , Mayrink Wilson , Genaro Odair , Tavares Carlos Alberto Pereira 
Published: November 2002

ABSTRACTAn antigen (LMS) prepared fromLeishmania major-like promastigotes was used in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of human and dog visceral leishmaniasis. The results were compared with those from the indirect immunofluorescent antibody test (IFAT). A total of 1,822 canine sera were tested, including sera from dogs with visceral leishmaniasis, transmissible venereal tumors, ehrlichiosis, rickettsiosis, or Chagas' disease and sera from healthy dogs. The antigen was also tested with 227 samples of human sera, including sera from patients with visceral, cutaneous, or diffuse cutaneous leishmaniasis and from noninfected individuals, as well as sera from patients with Chagas' disease, toxoplasmosis, rickettsiosis, hepatitis B, schistosomiasis, ascaridiasis, malaria, rheumatoid factor, leprosy and rheumatoid factor, tuberculosis, or leprosy. All dogs and all human patients had a clinical and/or serological and/or parasitological diagnosis. For detecting antibodies in sera from dogs with leishmaniasis, the antigen showed a sensitivity of 98%, specificity of 95%, and concordance of 93% and when used for detecting antibodies in human sera presented a sensitivity of 92%, specificity of 100%, and concordance of 92%. Comparison between ELISA and IFAT demonstrated that ELISA using the LMS antigen yielded more reliable results than IFAT. The LMS antigen displayed no cross-reactivity with sera from patients or dogs that had any of the other diseases tested.
Comparison ofStaphylococcus aureusIsolates from Bovine and Human Skin, Milking Equipment, and Bovine Milk by Phage Typing, Pulsed-Field Gel Electrophoresis, and Binary Typing
Authors: Zadoks R. N. , van Leeuwen W. B. , Kreft D. , Fox L. K. , Barkema H. W. , Schukken Y. H. , van Belkum A. 
Published: October 2002

ABSTRACTStaphylococcus aureusisolates (n= 225) from bovine teat skin, human skin, milking equipment, and bovine milk were fingerprinted by pulsed-field gel electrophoresis (PFGE). Strains were compared to assess the role of skin and milking equipment as sources ofS. aureusmastitis. PFGE ofSmaI-digested genomic DNA identified 24 main types and 17 subtypes among isolates from 43 herds and discriminated between isolates from bovine teat skin and milk. Earlier, phage typing (L. K. Fox, M. Gershmann, D. D. Hancock, and C. T. Hutton, Cornell Vet. 81:183-193, 1991) had failed to discriminate between isolates from skin and milk. Skin isolates from humans belonged to the same pulsotypes as skin isolates from cows. Milking equipment harbored strains from skin as well as strains from milk. We conclude thatS. aureusstrains from skin and from milk can both be transmitted via the milking machine, but that skin strains are not an important source of intramammaryS. aureusinfections in dairy cows. A subset of 142 isolates was characterized by binary typing with DNA probes developed for typing of humanS. aureus. Typeability and overall concordance with epidemiological data were lower for binary typing than for PFGE while discriminatory powers were similar. Within several PFGE types, binary typing discriminated between main types and subtypes and between isolates from different herds or sources. Thus, binary typing is not suitable as replacement for PFGE but may be useful in combination with PFGE to refine strain differentiation.
Molecular Epidemiology of Pneumococcal Carriage among Children with Upper Respiratory Tract Infections in Hanoi, Vietnam
Authors: Bogaert D. , Ha N. T. , Sluijter M. , Lemmens N. , de Groot R. , Hermans P. W. M. 
Published: October 2002

ABSTRACT

To investigate the molecular epidemiology of pneumococcal nasopharyngeal carriage in Hanoi, Vietnam, we studied 84 pneumococcal strains retrieved from children with upper respiratory tract infections. Serotypes 23F (32%), 19F (21%), 6B (13%), and 14 (10%) were found most often. A significant number of strains were antibiotic resistant. Fifty-two percent of the strains were (intermediate) resistant to penicillin, 87% were (intermediate) resistant to co-trimoxazole, 76% were resistant to tetracycline, 73% were resistant to erythromycin, and 39% were (intermediate) resistant to cefotaxime. Seventy-five percent were resistant to three or more classes of antibiotics. A high degree of genetic heterogeneity among the penicillin resistance genes was observed. In addition, the tetracycline resistance gene
tet
(M) and the erythromycin resistance gene
erm
(B) were predominantly observed among the isolates. Molecular analysis of the 84 isolates by restriction fragment end labeling (RFEL) revealed 35 distinct genotypes. Twelve of these genotypes represented a total of eight genetic clusters with 61 isolates (73%). The two largest clusters contained 24 and 12 isolates, and the isolates in those clusters were identical to the two internationally spreading multidrug-resistant clones Spain 23F-1 and Taiwan 19F-14, respectively. The remaining RFEL types were Vietnam specific, as they did not match the types in our reference collection of 193 distinct RFEL types from 16 countries. Furthermore, 57 of the 61 horizontally spreading isolates (93%) in the eight genetic clusters were covered by the seven-valent conjugate vaccine, whereas this vaccine covered only 43% of the isolates with unique genotypes. According to the serotype distribution of the nasopharyngeal pneumococcal isolates, this study suggests a high potential benefit of the seven-valent pneumococcal conjugate vaccine for children in Hanoi.
Increased Risk of Parvovirus B19 Infection in Young Adult Cancer Patients Receiving Multiple Courses of Chemotherapy
Authors: Kuo Sung-Hsin , Lin Liang-In , Chang Chee-Jen , Liu Yun-Ru , Lin Kuo-Sin , Cheng Ann-Lii 
Published: October 2002

ABSTRACTAn increased human parvovirus B19 infection rate has been observed in immunocompromised hosts. In this study, we sought to determine the prevalence of parvovirus B19 infection in adult cancer patients receiving multiple courses of systemic chemotherapy. From March 1999 through April 2000, 59 men and 68 women, with a median age of 49 (18 to 79) years, were enrolled in this study. They had received an average of 7.1 (4 to 32) courses of systemic chemotherapy. The median duration from the date of starting chemotherapy to the date of blood sampling was 11 (4 to 88) months. Serum B19 immunoglobulin G (IgG) and IgM levels were examined by an enzyme-linked immunosorbent assay, and B19 DNA was examined by a nested PCR. A group of 400 healthy blood donors served as the control group. The overall prevalences of anti-B19 IgG in adult cancer patients and healthy blood donors were 61.4 and 25.0%, respectively (P< 0.01). Anti-B19 IgM and B19 DNA were not detectable in these anti-B19 IgG-seropositive individuals. A further age-stratified comparison revealed that only patients younger than 40 years had a significantly higher anti-B19 IgG seropositivity rate than the controls (19 of 39 versus 53 of 310;P< 0.001). The increased prevalence of B19 infection in these 39 adult patients younger than 40 years might be clinically significant, since unexplained anemia, a common sequela of B19 infection, was detected in 3 of 20 seronegative patients (15.0%) and in 12 of 19 seropositive patients (63.2%) (P< 0.005). The results of this study suggest that adult patients younger than 40 years and receiving multiple courses of systemic chemotherapy may have a significantly increased risk of B19 infection. Prospective studies to define the time course and clinical consequence of B19 infection in this group of patients are needed.
Alafosfalin as a Selective Agent for Isolation of
Salmonella
from Clinical Samples
Authors: Perry J. D. , Riley G. , Gould F. K. , Perez J. M. , Boissier E. , Ouedraogo R. T. , Freydière A. M. 
Published: October 2002

ABSTRACT

The selectivity of a range of culture media for the detection of
Salmonella
was assessed using 435 strains of gram-negative bacteria. These media showed limited ability to inhibit non-
Salmonella
strains found in stool samples. We report the evaluation of alafosfalin as a selective agent for isolation of
Salmonella
from stool samples. Susceptibility studies with this agent showed that non-
typhi Salmonella
strains were relatively resistant (mean MIC, 10.2 mg/liter) compared to many coliforms including
Escherichia coli
(mean MIC, 0.7 mg/liter). A chromogenic medium, ABC medium, was modified to incorporate alafosfalin and was compared with standard ABC medium and Hektoen enteric agar for the isolation of
Salmonella
from 1,000 stool samples. On direct culture, modified ABC medium showed higher recovery of
Salmonella
(53.6%) compared with either ABC medium (35.7%) or Hektoen enteric agar (48.2%). We conclude that alafosfalin is a useful selective agent for the isolation of
Salmonella
from stool samples.
Rates of Detection of
Neisseria meningitidis
in Tonsils Differ in Relation to Local Incidence of Invasive Disease
Authors: Greiner Oliver , Berger Christoph , Day Philip J. R. , Meier Gabriela , Tang Christoph M. , Nadal David 
Published: October 2002

ABSTRACT

Nasopharyngeal swabbing substantially underestimates carriage of
Neisseria meningitidis
. Real-time PCR assays were employed to examine the presence of a broad range of bacteria and of
N. meningitidis
groups B and C, respectively, in tonsils from 26 individuals from Oxford, England, and 72 individuals from Zurich, Switzerland. The detection limit of each PCR system was DNA from one bacterial cell per reaction mixture. Tonsillar DNA did not inhibit amplification of meningococcal gene sequences, and
N. meningitidis
was detected in tonsils exposed to the bacterium. Whereas in both sets of patients other bacteria were detected,
N. meningitidis
group B and group C were only found in tonsils from Oxford where the incidence of invasive meningococcal disease is much higher than in Zurich. These observations suggest that PCR-based methods could be used for the detection of meningococcal carriage and that difference in disease incidence could be explained by different transmission rates in the community rather than host genetics or coexisting infections.
Real-Time PCR Method for Detection ofEncephalitozoonintestinalisfrom Stool Specimens
Authors: Wolk D. M. , Schneider S. K. , Wengenack N. L. , Sloan L. M. , Rosenblatt J. E. 
Published: October 2002

ABSTRACTThe prevalence of microsporidiosis is likely underestimated due to the labor-intensive, insensitive, and nonspecific clinical laboratory methods used for the diagnosis of this disease. A real-time PCR assay was designed to assess DNA extraction methods and to detect threeEncephalitozoonspecies in feces. Modifications of the MagNA Pure LC DNA isolation kit protocol (Roche Applied Sciences, Indianapolis, Ind.) were compared by using the automated MagNA Pure LC instrument (Roche) and fecal specimens spiked withEncephalitozoonintestinalisspores. Extracted DNA was amplified by the LightCycler (Roche) PCR assay. Assay sensitivity, reproducibility, and efficiency were assessed by comparing threshold crossover values achieved with different extraction and storage conditions (fresh, refrigerated, frozen, and preserved specimens). Optimal extraction conditions were achieved by using a commercial buffer, tissue lysis buffer (Roche), as the specimen diluent. LightCycler PCR results were compared to those obtained from routine stool microscopy with trichrome blue stain. The lower limit of detection for the LightCycler PCR assay varied by storage conditions from 102to 104spores/ml of feces, a value which represented a significant improvement over that achieved by staining (≥1.0 × 106spores/ml). Melting temperature analysis of the amplicons allowed for the differentiation of threeEncephalitozoonspecies (E.intestinalis,E.cuniculi, andE.hellem). The assay is readily adaptable to the clinical laboratory and represents the first real-time PCR assay designed to detectEncephalitozoonspecies.
Validation of Performance of the Gen-Probe Human Immunodeficiency Virus Type 1 Viral Load Assay with Genital Swabs and Breast Milk Samples
Authors: DeVange Panteleeff Dana , Emery Sandra , Richardson Barbra A. , Rousseau Christine , Benki Sarah , Bodrug Sharon , Kreiss Joan K. , Overbaugh Julie 
Published: October 2002

ABSTRACT
Human immunodeficiency type 1 (HIV-1) continues to spread at an alarming rate. The virus may be transmitted through blood, genital secretions, and breast milk, and higher levels of systemic virus in the index case, as measured by plasma RNA viral load, have been shown to correlate with increased risk of transmitting HIV-1 both vertically and sexually. Less is known about the correlation between transmission and HIV-1 levels in breast milk or genital secretions, in part because reliable quantitative assays to detect HIV-1 in these fluids are not available. Here we show that the Gen-Probe HIV-1 viral load assay can be used to accurately quantify viral load in expressed breast milk and in cervical and vaginal samples collected on swabs. Virus could be quantified from breast milk and swab samples spiked with known amounts of virus, including HIV-1 subtypes A, C, and D. As few as 10 copies of HIV-1 RNA could be detected above background threshold levels in ≥77% of assays performed with spiked breast milk supernatants and mock swabs. In genital swab samples from HIV-1-infected women, similar levels of HIV-1 RNA were consistently detected in duplicate swabs taken from the same woman on the same clinic visit, suggesting that the RNA values from a single swab sample can be used to measure genital viral load.
“
Actinobaculum massiliae
,” a New Species Causing Chronic Urinary Tract Infection
Authors: Greub Gilbert , Raoult Didier 
Published: October 2002

ABSTRACT

We report on a new
Actinobaculum
species, “
Actinobaculum massiliae
,” isolated from the urine of an elderly woman with recurrent cystitis. Its phenotypic pattern was similar to those of both of the other
Actinobaculum
species described to date. On 16S rRNA sequencing, the Marseille isolate shared 95% homology with
Actinobaculum suis
, 92 to 93% homology with
Actinobaculum schaalii
, 91 to 92% homology with
Arcanobacterium
spp., and 87 to 90% homology with
Actinomyces
species. A bootstrap value of 99% supports the node separating the
Actinobaculum
sp. from its closest neighbor (
A. suis
). In conclusion, on the basis of phenotypic, genotypic, and phylogenetic assessments, we show that the Marseille isolate is a previously unrecognized organism within the
Actinobaculum
genus, and we propose placement of the organism in the taxon “
Actinobaculum massiliae
.”
Dissemination of
Streptococcus pneumoniae
Clone Colombia
5
-19 in Latin America
Authors: Gamboa Liliana , Camou Teresa , Hortal María , Castañeda Elizabeth 
Published: October 2002

ABSTRACT

Streptococcus pneumoniae
serotype 5 is the third most common capsular type causing invasive diseases in children younger than 5 years in Latin America. Preliminary data on Colombian serotype 5 isolates indicated a common clonal origin associated with resistance to tetracycline (TET) and chloramphenicol (CHL). We studied 172
S. pneumoniae
serotype 5 invasive isolates from Argentina, Brazil, Colombia, Guatemala, Mexico, and Uruguay and confirmed the presence of the Colombia
5
-19 clone throughout Latin America. Fifteen subtypes of a pulsed-field gel electrophoresis pattern and 4 electrophoretic types (ET) were obtained. Most of the isolates from different geographical regions belonged to pattern A (34.3%), subtype A5 (41.9%), and ET1 (91.1%). The A pattern (
n
= 59) was resistant to TET and had variable resistance to CHL; it was present in Brazil (10.2%), Colombia (78%), Guatemala (8.5%), and Mexico (3.4%). Subtype A5 with variable susceptibility to TET and sensitive to CHL was found in Argentina (29.2%), Mexico (8.3%), and Uruguay (62.5%). Subtypes A1-A4, A7-A8, and A9-A11 (closely related to A) also shared ET1, while subtype A6 was assigned to ET1, ET2, and ET3. Eleven subtypes (
n
= 21) were found to be specific for one country each. In summary, the
S. pneumoniae
serotype 5 isolates from Latin American are genetically closely related but show different patterns of antibiotic resistance, probably as a result of horizontal transfer.
Both Urinary and Rectal
Escherichia coli
Isolates Are Dominated by Strains of Phylogenetic Group B2
Authors: Zhang Lixin , Foxman Betsy , Marrs Carl 
Published: October 2002

ABSTRACT

To compare the genetic structures of uropathogenic and commensal
Escherichia coli
populations, a total of 181 urinary and rectal
E. coli
isolates were classified into intraspecies phylogenetic groups by PCR amplifications of phylogenetic markers. The genetic variability of these isolates within phylogenetic groups was further assessed by enterobacterial repetitive intergenic consensus (ERIC) typing. The distributions of 10 known virulence factors were also examined. In contrast with most reports, phylogenetic group B2 not only accounted for the majority of urinary isolates from young women with urinary tract infections (69%) but also was the dominant group among the rectal isolates from healthy young women (48%). Such difference may be explained by geographic variation, difference in host population characteristics, or differences in sampling method, or a combination of the three. Strains with known virulence factors most frequently belonged to phylogenetic groups B2 and D. Additionally, group B2 and D rectal isolates were more heterogeneous than urinary isolates. Two subclusters existed within group B2 strains by ERIC typing. These subclusters were not evenly distributed between rectal and urine isolates and differed in virulence gene distribution.
Protocol for Real-Time PCR Identification of Anthrax Spores from Nasal Swabs after Broth Enrichment
Authors: Oggioni Marco R. , Meacci Francesca , Carattoli Alessandra , Ciervo Alessandra , Orru Germano , Cassone Antonio , Pozzi Gianni 
Published: October 2002

ABSTRACT

A mass-screening protocol for the diagnosis of anthrax from nasal swabs based on an enrichment step in liquid medium was devised. Incubation for growth was performed in autoclavable vials and racks which allow real-time PCR analysis of sterilized cultures. A dual-color PCR was set up with primers and probes for the chromosomal marker
rpoB
and the plasmid marker
lef
. Specific primer and probe sets were designed for the differentiation of
Bacillus anthracis
from
B. cereus
and for the differentiation of the Sterne vaccine strain from field isolates and the Ames strain, which was used in the recent anthrax bioterrorist attack. The present protocol thus combines the high specificity and sensitivity of real-time PCR with excellent biosafety and the low hands-on time necessary for the processing of large numbers of samples, which is extremely important during control programs involving the processing of large numbers of samples.
Extrapulmonary and Pulmonary Tuberculosis in Antananarivo (Madagascar): High Clustering Rate in Female Patients
Authors: Rasolofo Razanamparany Voahangy , Ménard Didier , Aurégan Guy , Gicquel Brigitte , Chanteau Suzanne 
Published: October 2002

ABSTRACT

Antananarivo, the capital city of Madagascar, has an endemic focus of tuberculosis (TB). We specifically studied patients with extrapulmonary TB (EPTB) and grouped patients according to infected body site. The strains were characterized by IS
6110
fingerprinting and compared with those isolated from patients with pulmonary TB (PTB) during the same period in order to determine the possible association between the genotype and the clinical expression of TB. A total of 316 TB patients were included in this study: 151 individuals with EPTB, 10 with both PTB and EPTB, and 155 with PTB alone. Pleural TB was the major EPTB localization (77%) and was found more often in older patients, while PTB or EPTB in which the localization was other than pleural (other EPTB) was found in younger patients. The male-to-female ratio was slightly higher in pleural TB patients (3.06:1) than in patients with other EPTB (1.35:1). There was no significant difference in the BCG status among patients with PTB, pleural TB, and other EPTB. Analysis of IS
6110
patterns showed that 167 patients (52.8%) were assigned to 37 clusters of 2 to 34 patients. Analysis of the IS
6110
clusters and the IS
6110
families did not show any association with a particular clinical expression of the disease. Patients with PTB or other EPTB were more likely to have strains with one IS
6110
copy than patients with pleural TB. The clustering rate was found to be significantly higher in female patients (62%) than in male patients (48%) (
P
= 0.029), suggesting that Malagasy women were more likely to progress to disease after infection than men.
mecA
Gene Is Widely Disseminated in
Staphylococcus aureus
Population
Authors: Wielders C. L. C. , Fluit A. C. , Brisse S. , Verhoef J. , Schmitz F. J. 
Published: October 2002

ABSTRACT

Methicillin-resistant
Staphylococcus aureus
(MRSA) is one of the most important causes of hospital infections worldwide. High-level resistance to methicillin is caused by the
mecA
gene, which encodes an alternative penicillin-binding protein, PBP 2a. To determine the clonal relationships between methicillin-susceptible
S. aureus
(MSSA) and MRSA, we typed 1,069
S. aureus
isolates (493 MSSA isolates and 576 MRSA isolates), collected mainly in North American and European hospitals between the 1960s and the year 2000, using pulsed-field gel electrophoresis and ribotyping. Of 10 widespread
S. aureus
lineages recognized, 8 had corresponding
mecA
-positive strains. Multiresistant MRSA strains are found in hospitals worldwide, while unrelated and more susceptible strains represent less than 1% of the MRSA population. This supports the hypothesis that horizontal transfer plays an important role in the dissemination of the
mecA
gene in the
S. aureus
population.
Multicenter Evaluation of Ethambutol Susceptibility Testing of
Mycobacterium tuberculosis
by Agar Proportion and Radiometric Methods
Authors: Madison B. , Robinson-Dunn B. , George I. , Gross W. , Lipman H. , Metchock B. , Sloutsky A. , Washabaugh G. , Mazurek G. , Ridderhof J. 
Published: October 2002

ABSTRACT

Reproducibility of ethambutol (EMB) susceptibility test results for
Mycobacterium tuberculosis
has always been difficult for a variety of reasons, including the narrow range between the critical breakpoint for EMB resistance and the MIC for susceptible strains, borderline results obtained with the BACTEC 460TB method, the presence of microcolonies determined using the agar proportion (AP) method, and a lack of agreement between these two testing methods. To assess the frequency of these problems,
M. tuberculosis
drug susceptibility data were collected in a multicenter study involving four laboratories. Resistant, borderline, and susceptible isolates were shared among the laboratories to measure interlaboratory test agreement. Half of isolates determined by BACTEC 460TB to be resistant were determined to be susceptible by the AP method. Isolates determined to be resistant to EMB by both BACTEC 460TB and AP methods were almost always resistant to isoniazid. Results from isolates tested by the BACTEC 460TB method with an EMB concentration of 3.75 μg/ml in addition to the standard 2.5 μg/ml did not show improved agreement by the AP method. While these results do not indicate that the AP method is more accurate than the BACTEC 460TB method, laboratories should not report EMB monoresistance based on BACTEC 460TB results alone. Monoresistance to EMB should only be reported following confirmation by the AP method. Microcolonies could not be confirmed as resistant by the BACTEC 460TB method or by repeat testing with the AP method and do not appear to be indicative of resistance.