bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of
Paracoccidioides brasiliensis
gp70 Circulating Antigen and Follow-Up of Patients Undergoing Antimycotic Therapy
Authors: Marques da Silva Silvia Helena , de Mattos Grosso Daniela , Lopes José Daniel , Colombo Arnaldo Lopes , Blotta Maria Heloisa Souza Lima , Queiroz-Telles Flávio , Pires de Camargo Zoilo 
Published: October 2004

ABSTRACT

Paracoccidioidomycosis (PCM), one of the most important systemic mycoses in Central and South America, is caused by the dimorphic fungus
Paracoccidioides brasiliensis
and has a high prevalence in Brazil. Glycoproteins of 43 and 70 kDa are the main antigenic compounds of
P. brasiliensis
and are recognized by Western blotting by 100 and 96% of PCM patient sera, respectively. In the present study, an inhibition enzyme-linked immunosorbent assay (ELISA) was used to detect gp70 in different biological samples from patients with PCM. gp70 was detected in 98.76% of 81 serum samples, with an average concentration of 8.19 μg/ml. The test was positive for 100% of the patients with the acute and chronic unifocal forms of PCM and 98.43% of the patients with the multifocal chronic form, with average concentrations of 11.86, 4.83, and 7.87 μg/ml, respectively. Bronchoalveolar lavage fluid from 23 patients with pulmonary unifocal PCM and 14 samples of cerebrospinal fluid from patients with neurological PCM were also tested for gp70 detection, with the test showing 100% sensitivity and 100% specificity, with mean gp70 concentrations of 7.5 and 6.78 μg/ml, respectively. To investigate the potential of gp70 detection by inhibition ELISA for the follow-up of PCM patients during antimycotic therapy with itraconazole (ITZ), the sera of 23 patients presenting with the chronic multifocal form of PCM were monitored at regular intervals of 1 month for 12 months. The results showed a decrease in circulating gp70 levels during treatment which paralleled the reduction in anti-
P. brasiliensis
antibody levels. The detection of
P. brasiliensis
gp70 from the biological fluids of patients suspected of having PCM proved to be a promising method for diagnosing infection and evaluating the efficacy of ITZ treatment.
Multiple-Locus Variable-Number Tandem Repeat Analysis, a Novel Typing Scheme To Study the Genetic Relatedness and Epidemiology of
Enterococcus faecium
Isolates
Authors: Top Janetta , Schouls Leo M. , Bonten Marc J. M. , Willems Rob J. L. 
Published: October 2004

ABSTRACT

Multiresistant
Enterococcus faecium
is a major cause of hospital acquired infections and outbreaks. Here, we describe the development of multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) as a novel typing method to assess the genetic relatedness of
E. faecium
isolates. Six VNTR loci were used to genotype 392 isolates recovered from different animals and human community, hospital survey, and clinical isolates. From 3 to 13 alleles were found per locus, resulting in 127 different MLVA profiles. Clustering of MLVA profiles confirmed the host-specific genogroups found by multilocus sequence typing (MLST) and showed the grouping of clinical and epidemic isolates that belonged to the MLST-C1 cluster in a distinct MLVA-C1 cluster (sensitivity of 97% and specificity of 90%). Furthermore, the discriminatory power of MLVA is comparable to MLST. MLVA profiles appeared to be relatively stable, since isolates from a single outbreak shared the same MLVA profile, which is a prerequisite when MLVA is used to study hospital outbreaks. Our data show that MLVA is a highly reproducible and portable typing method; in contrast to MLST, it is fast, relatively cheap, and easy to perform. Furthermore, it has the abilities of MLST to recognize genetically related and potential epidemic isolates. Submission of MLVA profiles is possible via a Web-based database for international comparison.
Development and Evaluation of a Chromogenic Agar Medium for Methicillin-Resistant
Staphylococcus aureus
Authors: Perry John D. , Davies Amie , Butterworth Lynne A. , Hopley Andrew L. J. , Nicholson Audrey , Gould F. Kate 
Published: October 2004

ABSTRACT

We describe here the development and evaluation of MRSA ID, a new chromogenic agar medium for the specific isolation and identification of methicillin-resistant
Staphylococcus aureus
(MRSA). We used S. aureus ID (bioMérieux, La Balme Les Grottes, France) and supplemented it with various antimicrobials, including cefoxitin, ciprofloxacin, oxacillin, and methicillin. Cefoxitin proved to be superior to the other antimicrobials for the selection of MRSA from other strains of
S. aureus
. MRSA ID (consisting of S. aureus ID supplemented with 4 mg of cefoxitin/liter) was evaluated by the use of 747 swabs from various clinical sites. All specimens were also cultured on CHROMagar MRSA and oxacillin resistance screening agar base (ORSAB) and in selective mannitol broth (SMB). A total of 85 MRSA strains were isolated by a combination of all methods. After 22 to 24 h of incubation, 80% of the MRSA strains were isolated as green colonies on MRSA ID, compared with 59 and 62% of the strains that were isolated as colored colonies on CHROMagar MRSA and ORSAB, respectively. After 48 h of incubation, 89, 72, and 78% of the MRSA strains were isolated on MRSA ID, CHROMagar MRSA, and ORSAB, respectively. Sixty-five percent of the strains were isolated by growth in SMB. The specificities of MRSA ID, CHROMagar MRSA, ORSAB, and SMB were 99.5, 99.3, 97.9, and 92.8%, respectively, after 22 to 24 h of incubation. We conclude that MRSA ID is a sensitive and specific medium for the isolation and identification of MRSA.
Longitudinal Study of Viruses Associated with Canine Infectious Respiratory Disease
Authors: Erles Kerstin , Dubovi Edward J. , Brooks Harriet W. , Brownlie Joe 
Published: October 2004

ABSTRACTIn this investigation a population of dogs at a rehoming center was monitored over a period of 2 years. Despite regular vaccination of incoming dogs against distemper, canine adenovirus type 2 (CAV-2), and canine parainfluenza virus (CPIV), respiratory disease was endemic. Tissue samples from the respiratory tract as well as paired serum samples were collected for analysis. The development of PCR assays for the detection of CPIV, canine adenovirus types 1 and 2, and canine herpesvirus (CHV) is described. Surprisingly, canine adenovirus was not detected in samples from this population, whereas 19.4% of tracheal and 10.4% of lung samples were positive for CPIV and 12.8% of tracheal and 9.6% of lung samples were positive for CHV. As reported previously, a novel canine respiratory coronavirus (CRCoV) was detected in this population (K. Erles, C. Toomey, H. W. Brooks, and J. Brownlie, Virology310:216-223, 2003). Infections with CRCoV occurred mostly during the first week of a dog's stay at the kennel, whereas CPIV and CHV were detected at later time points. Furthermore, the evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to CPIV and an immunofluorescence assay for detection of antibodies to CHV is described. This study shows that CPIV is present at kennels despite vaccination. In addition, other agents such as CHV and CRCoV may play a role in the pathogenesis of canine respiratory disease, whereas CAV-2 and canine distemper virus were not present in this population, indicating that their prevalence in the United Kingdom is low due to widespread vaccination of dogs.
Nocardia mexicana
sp. nov., a New Pathogen Isolated from Human Mycetomas
Authors: Rodríguez-Nava Verónica , Couble Andrée , Molinard Claudie , Sandoval Horacio , Boiron Patrick , Laurent Frédéric 
Published: October 2004

ABSTRACT

Three isolates collected from human mycetomas and showing an unusual brownish purple pigmentation on Bennett agar plates were analyzed by a polyphasic taxonomic approach, including morphological, biochemical, physiological, and chemotaxonomic properties coupled with genomic and phylogenetic analysis. It clearly appeared that these microorganisms were distinct from their closest phenotypic and genetic match, the most related species according to 16S rRNA gene sequence analysis being
Nocardia pseudobrasiliensis.
The data obtained indicated that the three clinical strains should be recognized as a new species for which the name
Nocardia mexicana
sp. nov. is proposed.
High Rates of Clustering of Strains Causing Tuberculosis in Harare, Zimbabwe: a Molecular Epidemiological Study
Authors: Easterbrook Philippa J. , Gibson Andrea , Murad Shahed , Lamprecht Dianie , Ives Natalie , Ferguson Alex , Lowe Odette , Mason Peter , Ndudzo Angelika , Taziwa Alfred , Makombe Robert , Mbengeranwa Lovemore , Sola Christophe , Rostogi Nalim , Drobniewski Francis 
Published: October 2004

ABSTRACT
We examined the pattern of tuberculosis (TB) transmission (i.e., reactivation versus recent transmission) and the impact of human immunodeficiency virus (HIV) infection in Harare, Zimbabwe. Consecutive adult smear-positive pulmonary TB patients presenting to an urban hospital in Harare were enrolled. A detailed epidemiological questionnaire was completed, and tests for HIV type 1 and CD4 cell counts were performed for each patient. Molecular fingerprinting of the genomic DNA recovered from cultures of sputum was performed by two molecular typing methods: spacer oligonucleotide typing (spoligotyping) and analysis of variable number of tandem DNA repeats (VNTRs). A cluster was defined as isolates from two or more patients that shared the same spoligotype pattern or the same VNTR pattern, or both. DNA suitable for typing was recovered from 224 patients. The prevalence of HIV infection was 79%. Of 187 patient isolates (78.6%) typed by both spoligotyping and analysis of VNTRs, 147 were identified as part of a cluster by both methods. By spoligotyping alone, 84.1% of patient isolates were grouped into 20 clusters. The cluster size was generally <8 patient isolates, although three large clusters comprised 68, 25, and 23 patient isolates. A total of 89.4% of the patient isolates grouped into 12 clusters defined by analysis of VNTRs, with 2 large clusters consisting of 127 and 13 patient isolates, respectively. Thirty-six percent of patient isolates with a shared spoligotype and 17% with a shared VNTR pattern were geographically linked within Harare, but they were not linked on the basis of the patient's home district. In a multivariate analysis, there were no independent predictors of clustering, including HIV infection status. Comparison with the International Spoligotype database (Pasteur Institute, Pointe à Pitre, Guadeloupe) demonstrated that our three largest spoligotype clusters are well recognized and ubiquitous in Africa. In this epidemiologically well characterized urban population with a high prevalence of HIV infection, we identified a very high level of strain clustering, indicating substantial ongoing recent TB transmission. Geographic linkage could be detected in a proportion of these clusters. A small group of actively circulating strains accounted for most of the cases of TB transmission.
Controlled Clinical Comparison of the BacT/ALERT FN and the Standard Anaerobic SN Blood Culture Medium
Authors: Mirrett S. , Petti C. A. , Woods C. W. , Magadia R. , Weinstein M. P. , Reller L. B. 
Published: October 2004

ABSTRACT

To determine the optimal anaerobic companion bottle to pair with the BacT/ALERT (bioMérieux, Durham, N.C.) nonvented aerobic FA (FA) medium for recovery of pathogenic microorganisms from adult patients with bacteremia and fungemia, we compared the BacT/ALERT FN (FN) anaerobic bottle with the standard BacT/ALERT SN (SN) anaerobic bottle. Each bottle, FA, FN, and SN, was filled with 8 to 12 ml of blood. Of 11,498 blood culture sets received in the clinical microbiology laboratories at two university medical centers, 7,945 sets had all three bottles filled adequately and 8,569 had both anaerobic bottles filled adequately. Of 686 clinically important (based on previously published criteria) isolates detected in one or both adequately filled anaerobic bottles, more staphylococci (
P
< 0.001), including
Staphylococcus aureus
(
P
< 0.001); members of the family
Enterobacteriaceae
(
P
< 0.001); and all microorganisms combined (
P
< 0.001) were detected in FN bottles. In contrast, more
Pseudomonas aeruginosa
isolates (
P
< 0.01) and yeasts (
P
< 0.001) were detected in SN bottles. More
Bacteroides fragilis
group bacteremias were detected only in the FN (six) than in the SN (one) anaerobic bottle (
P
= not significant). Overall, the mean time to detection was shorter with FN (16.8 h) than with SN (18.2 h). This difference in time to detection was greatest for the
B. fragilis
group: FN, 28 h, versus SN, 60.0 h. Many of the facultative microorganisms recovered in either FN or SN were also found in the companion FA. When microorganisms found in the companion FA bottle were omitted from the analysis, significantly more staphylococci (
P
< 0.001), including
S. aureus
(
P
< 0.001), and
Enterobacteriaceae
(
P
< 0.005) still were detected in FN bottles, whereas there were no significant differences for
P. aeruginosa
and yeasts, which were found as expected in FA bottles. We conclude that the companion anaerobic FN bottle detects more microorganisms than does the anaerobic SN bottle when used in conjunction with the nonvented aerobic FA bottle in the BacT/ALERT blood culture system.
Comparative Analysis of Gtf Isozyme Production and Diversity in Isolates of
Streptococcus mutans
with Different Biofilm Growth Phenotypes
Authors: Mattos-Graner Renata O. , Napimoga Marcelo H. , Fukushima Kasuo , Duncan Margaret J. , Smith Daniel J. 
Published: October 2004

ABSTRACT

Streptococcus mutans
is the main pathogenic agent of dental caries. Glucosyltransferases (Gtfs) produced by these bacteria are important virulence factors because they catalyze the extracellular synthesis of glucans that are necessary for bacterial accumulation in the dental biofilm. The diversity of GtfB and GtfC isozymes was analyzed in 44 genotypes of
S. mutans
that showed a range of abilities to form biofilms in vitro. Several approaches were used to characterize these isozymes, including restriction fragment length polymorphism analysis of the
gtfB
and
gtfC
genes, zymographic analysis of the identified GtfB and GtfC genotypes, and quantitation of isozyme production in immunoblot experiments with specific monoclonal antibodies. A high diversity of
gtf
genes, patterns of enzymatic activity, and isozyme production was identified among the isolates tested. GtfC and, to a lesser extent, GtfB were produced in significantly higher amounts by strains that had high biofilm-forming ability than by strains with low biofilm-forming ability. Biofilm formation was independent of the GtfB and GtfC genotype. Atypical strains that showed an apparent single Gtf isozyme of intermediate size between GtfB and GtfC were also identified. The results indicate that various expression levels of GtfB and GtfC isozymes are associated with the ability of distinct
S. mutans
genotypes to grow as biofilms, strengthening the results of previous genetic and biochemical studies performed with laboratory strains. These studies also emphasize the need to identify factors that control
gtf
gene expression.
Aqueous Humor and Serum Immunoblotting for Immunoglobulin Types G, A, M, and E in Cases of Human Ocular Toxoplasmosis
Authors: Garweg Justus G. , Garweg Silvia-Daniela L. , Flueckiger Franziska , Jacquier Patrick , Boehnke Matthias 
Published: October 2004

ABSTRACTThe purpose of this study was to compare the local and systemicToxoplasma-specific humoral immune responses in individuals with ocular toxoplasmosis (OT). To this end, paired aqueous humor and serum samples from 46 individuals with active OT and from 30 individuals without inflammatory eye disease (controls) were analyzed by immunoblotting for anti-Toxoplasmaimmunoglobulin G (IgG), IgA, IgM, and IgE directed against 20- to 120-kDa antigens. The presence in the aqueous humor of a unique band, or of at least three bands that were at least three times more intense in aqueous humor than in serum, was taken as evidence of local antibody production. IgG bands were detected in 98% of the aqueous humor samples, while IgA bands were detected in 76%, IgM bands were detected in 8%, and IgE bands were not detected in any. Evidence of local production of specific antibodies was found in 32 cases (70%) (IgG in 23 [50%]; IgA in 16 [35%]). In 10 instances (22%), routine laboratory tests were not indicative of OT. In 14 cases (30%), no local antibody production was detected by immunoblotting; 3 of these cases yielded evidence of local antibody production according to the Goldmann-Witmer coefficient. Local antibody production was revealed for 7 of the 30 controls (23%). Hence, the sensitivity of immunoblotting for IgG and IgA is 70%, and the specificity is 77%. We conclude that immunoblotting for local specific IgG and IgA supports the clinical diagnosis of OT in 70% of cases. In 22% of these, the diagnosis is not confirmed by other laboratory tests. Hence, immunoblotting increases the sensitivity of routine laboratory tests and should be considered for samples that register negative by such tests.
Seroprevalence of Antibodies against Noroviruses among Students in a Chinese Military Medical University
Authors: Dai Ying-chun , Nie Jun , Zhang Xu-fu , Li Zhi-feng , Bai Yang , Zeng Zhi-rong , Yu Shou-yi , Farkas Tibor , Jiang Xi 
Published: October 2004

ABSTRACT

Noroviruses (NVs) are important causes of nonbacterial gastroenteritis in humans, but the role of NVs as a cause of diseases in the Chinese people, particularly in Chinese military personnel, remains unclear. This study investigated antibody prevalence and factors that associate with the prevalence of antibody to NVs among students attending a military medical university. Serum specimens were tested by an enzyme-linked immunosorbent assay for immunoglobulin G antibody to recombinant capsid antigens of three NVs (rNorwalk, rMxV, and rVA387). Of 588 serum samples tested, the antibody prevalence was 88.9, 54.1, or 90.0% for the three antigens, respectively. There were significant differences in the prevalence of antibody to rMxV between blood types (
P
< 0.05); the prevalence for type O was the highest (62.5%), and the prevalence for type B was the lowest (49.1%). The average optical density values for antibody to rNorwalk and rMxV were lowest among students with type B. The number of students who did not have antibody to any of the three antigens was the highest for blood type B (6.9%) compared to other blood types (0.8 to 3.4% [
P
< 0.006]). The antibody prevalence also varied with the hometown residencies of the students before joining the military, with the highest rates for students from rural areas, lower rates for students from small towns or villages, and the lowest rates for students from large cities. The numbers of students who did not have antibody to any of the three antigens were highest for students from the large cities, lower for students from small towns or villages, and lowest for students from rural areas. The distribution of ABO blood types did not differ among the three groups. These data suggest that NVs are prevalent in China and that both genetic and environmental factors play a role in NV infection.
Longitudinal Study of Transmission, Diversity, and Stability of
Streptococcus mutans
and
Streptococcus sobrinus
Genotypes in Brazilian Nursery Children
Authors: Klein Marlise Inêz , Flório Flávia Martão , Pereira Antonio Carlos , Höfling José Francisco , Gonçalves Reginaldo Bruno 
Published: October 2004

ABSTRACT

The aim of this study was to perform a follow-up evaluation of the
Streptococcus mutans
and
Streptococcus sobrinus
colonization profile of children's oral cavities, which included the pattern of vertical transmission from mother to child, genotypic diversity, and stability of the strains. The subjects were 16 mother-child pairs, who were monitored for 20 months. Samples of saliva, tongue dorsum, alveolar ridge mucosa, and dental plaque from the children were collected bimonthly. Saliva samples from the mothers were also collected. After isolation and identification, the arbitrarily primed PCR method was performed for the genotypic characterization of
S. mutans
(968 isolates) and
S. sobrinus
(111 isolates). At the time the strains were acquired, the children harbored one to four distinct genotypes of
S. mutans
and only one genotype of
S. sobrinus
. Although
S. mutans
prevalence and genotypic diversity were greater than those of
S. sobrinus
, the presence of matching genotypes of
S. mutans
and
S. sobrinus
was similar (in 81.25 and 83.33% of mother-child pairs, respectively), suggesting vertical transmission for both species. This longitudinal study showed an increase in genotypic diversity of
S. mutans
in the oral cavity during the follow-up period: most of the initially acquired genotypes persisted, normally those genotypes transmitted by the mother, and some were lost during follow-up; new strains were also acquired. In conclusion,
S. mutans
and
S. sobrinus
genotypes acquired from maternal or alternative sources may show effective persistence in the oral cavity and/or transitory detection in the children's mouths, reflecting the continuous development of oral microbiota in children.
Prospective Study of Human Metapneumovirus Infection in Children Less Than 3 Years of Age
Authors: König Brigitte , König Wolfgang , Arnold Ralf , Werchau Herrmann , Ihorst Gabriele , Forster Johannes 
Published: October 2004

ABSTRACT

Most lower respiratory tract infections (LRTIs) in children under the age of 3 years are due to respiratory syncytial virus (RSV). Epidemiological, host, and viral factors eventually account for the severity of LRTIs, but they do not completely explain it. Human metapneumovirus (hMPV) was recently identified in children with LRTIs. In a population-based prospective multicenter study (the PRI.DE study, conducted in Germany over 2 years), we tested 3,369 nasopharyngeal secretions from children younger than 3 years of age with LRTIs for RSV A and B, influenza viruses (IVs) A and B, and parainfluenza viruses (PIVs) 1 to 3. Of the children requiring intensive care (
n
= 85), 18% had hMPV infections, and 60% of these children were infected with hMPV in combination with RSV. We did not detect hMPV in a randomly selected subset of RSV-positive nasopharyngeal secretions (
n
= 120) from children not requiring intensive care support. hMPV was detected in <1% of virus-negative samples from patients without intensive care support (
n
= 620). Our data support the hypothesis that coinfections with RSV and hMPV are more severe than infections with either RSV or hMPV alone, at least in children younger than 3 years of age.