bluuebunny/crossref_metadata_2025_split huggingface dataset:
Coinfection of Enteric
Helicobacter
spp. and
Campylobacter
spp. in Cats
Authors: Shen Z. , Feng Y. , Dewhirst F. E. , Fox J. G. 
Published: July 2002

ABSTRACT

During a 6-year period, 64 of 227 commercially reared cats had microaerobic bacteria isolated from their feces. All the isolates were initially identified as
Campylobacter
-like organisms based on biochemical and phenotypic characteristics. DNA extractions from 51 of these isolates were subjected to PCR using primers specific for
Helicobacter
spp. and
Campylobacter
spp. Of the isolates, 92% (47 of 51 isolates) were positive for
Campylobacter
spp., 41% (21 of 51 isolates) were positive for
Helicobacter
spp., 33% (17 of 51 isolates) were positive for both genera, 59% (30 of 51 isolates) were positive only for
Campylobacter
spp., and 8% (4 of 51) were positive only for
Helicobacter
spp. Sixteen of the 47
Campylobacter
-positive cultures were positive for more than one
Campylobacter
spp. Based on a species-specific PCR assay, 83% of the isolates were identified as
Campylobacter helveticus
, 47% of the isolates were identified as
Campylobacter upsaliensis,
and 6% of the isolates were classified as
Campylobacter jejuni
. The 1.2-kb PCR products of the 16S rRNA genes of 19
Helicobacter
species isolates were subjected to restriction fragment length polymorphism (RFLP) analysis. Of the five different RFLP patterns obtained, two clustered with
Helicobacter
(“
Flexispira
”) taxon 8, one clustered with
Helicobacter bilis
, one clustered with
Helicobacter canis
, and the remaining pattern was closely related to a novel
Helicobacter
sp. strain isolated from a woodchuck. The sequence data for the 16S rRNA genes of 10
Helicobacter
spp. validated the RFLP-based identification of these isolates. This study demonstrated that biochemical and phenotypic characteristics of microaerobic organisms in cat feces were insufficient to characterize mixed
Helicobacter
and
Campylobacter
infections. Molecular structure-based diagnostics using genus- and species-specific PCR, RFLP analysis, and 16S rRNA sequence analysis enabled the identification of multiple microaerobic species in individual animals. The clinical relevance of enteric
Helicobacter
and
Campylobacter
coinfection in cats will require further studies.
Molecular Epidemiology Study of Exogenous Reinfection in an Area with a Low Incidence of Tuberculosis
Authors: Bandera Alessandra , Gori Andrea , Catozzi Lidia , Esposti Anna Degli , Marchetti Giulia , Molteni Chiara , Ferrario Giulio , Codecasa Luigi , Penati Valeria , Matteelli Alberto , Franzetti Fabio 
Published: July 2002

ABSTRACT

In geographical areas with a low incidence of tuberculosis, recurrent tuberculosis is generally due to reactivation of the disease. However, the relative contribution of tuberculosis reinfection increases in parallel with the incidence of disease and is likely to depend on the epidemiological context: factors such as the spread of multidrug resistance, human immunodeficiency virus (HIV) infection, and immigration from developing countries could modify disease transmission in areas at low risk for tuberculosis. A molecular epidemiology study was performed in Lombardy, Northern Italy, where the incidence of tuberculosis is 17.5 cases per 100,000 persons. A total of 2,452 cases of culture-confirmed tuberculosis in 2,127 patients were studied. A group of 32 patients (1.5%), each of whom had two episodes of tuberculosis with cure as the outcome of the first episode and with more than 6 months between the two episodes, were studied by means of restriction fragment length polymorphism DNA fingerprinting analysis. For 5 of the 32 patients (16%), the DNA fingerprinting patterns of
Mycobacterium tuberculosis
strains responsible for the second episode did not match those of the corresponding isolates of the first episode, indicating exogenous reinfection. Two of these patients developed multidrug-resistant tuberculosis during the second episode, and in three cases the isolates belonged to clusters of
M. tuberculosis
strains spreading in the community. A fourfold-increased risk for reinfection was observed in immigrant patients compared to Italian subjects. In contrast, a higher risk of relapse rather than reinfection was evidenced in HIV-positive subjects and in patients infected with multidrug-resistant tuberculosis. Episodes of tuberculosis reinfection in areas with a low incidence of tuberculosis are rare compared to those in high-incidence geographical regions. In populations that have immigrated from high-risk areas, reinfection may represent a considerable contributor to the rate of recurrent tuberculosis. This finding emphasizes the importance of containing the spread of epidemic strains in close communities, in order to prevent changes in global tuberculosis trends for developed countries.
Phaeohyphomycotic Cyst Caused byColletotrichum crassipes
Authors: Martins Castro Luiz Guilherme , da Silva Lacaz Carlos , Guarro Josep , Gené Josepa , Heins-Vaccari Elisabeth Maria , de Freitas Leite Roseli Santos , Hernández Arriagada Giovana Letı́cia , Ozaki Reguera Márcia Maria , Ito Eunice Miki , Valente Neusa Yuriko Sakai , Spina Nunes Ricardo 
Published: July 2002

ABSTRACTA case of phaeohyphomycosis is reported in a male renal transplant recipient with a nodular lesion in the right leg who was treated with immunosuppressing drugs. The lesion consisted of a purulent cyst with thick walls. The cyst was excised surgically, and the patient did not receive any antifungal therapy. One year later he remains well. Histological study of the lesion showed a granulomatous reaction of epithelioid and multinucleate giant cells, with a central area of necrosis and pus. Fontana-Masson staining demonstrated the presence of pigmented hyphal elements. The fungusColletotrichum crassipeswas grown in different cultures from the cyst. The in vitro inhibitory activities of eight antifungal drugs against the isolate were tested. Clotrimazole and UR-9825 were the most active drugs. This case represents the first known reported infection caused by this rare species.
Assessment, by Transcription-Mediated Amplification, of Virologic Response in Patients with Chronic Hepatitis C Virus Treated with Peginterferon α-2a
Authors: Sarrazin Christoph , Hendricks David A. , Sedarati Farhad , Zeuzem Stefan 
Published: July 2002

ABSTRACT
Transcription-mediated amplification (TMA) is an isothermal, autocatalytic target amplification method which has the potential to detect less than 50 hepatitis C virus (HCV) RNA copies/ml (10 IU/ml). The TMA assay was used to assess the presence of residual HCV RNA in plasma from patients treated with polyethylene glycol-modified interferon α-2a (peginterferon α-2a) who showed a virologic relapse after the end of therapy. Stored end-of-treatment and end-of-follow-up plasma samples from 177 of 267 patients treated with peginterferon α-2a (S. Zeuzem et al., N. Engl. J. Med. 343:1666–1672, 2000) were available for retesting by TMA. Plasma samples from patients in the same study who exhibited virologic relapse after treatment with standard interferon α-2a served as controls. Virologic response during the trial was defined as HCV RNA that was undetectable using a PCR-based test system with a sensitivity of 50 IU/mL (Cobas Amplicor HCV version 2.0) and was compared with TMA-based retesting results (VERSANT HCV RNA Qualitative Assay). Residual HCV RNA was detected in 4 of 60 cases (7%) by the TMA technology in end-of-treatment plasma samples from patients who relapsed after receiving peginterferon α-2a and in 6 of 18 patients (33%) following therapy with standard interferon α-2a. For peginterferon α-2a-treated patients with sustained virologic response, HCV RNA was detectable by TMA in end-of-treatment samples in 3 of 78 cases but in none of the end-of-follow-up samples. For all end-of-treatment and end-of-follow-up plasma samples of virologic nonresponders, a complete concordance between the PCR-based assay and TMA was observed. In conclusion, in patients with virologic relapse after the end of therapy, according to PCR, who were treated with peginterferon α-2a or standard interferon α-2a, residual HCV RNA was detectable in end-of-treatment samples by the TMA-based assay in 7 or 33% of cases, respectively. The lower rate of residual HCV RNA detection by TMA for patients treated with peginterferon α-2a than that for patients treated with standard interferon α-2a may be due to the maintained antiviral pressure of the long-acting peginterferon α-2a at the end-of-treatment visit.
Enhancing the Specificity of the COBAS AMPLICOR CT/NG Test for
Neisseria gonorrhoeae
by Retesting Specimens with Equivocal Results
Authors: Van Der Pol Barbara , Martin David H. , Schachter Julius , Quinn Thomas C. , Gaydos Charlotte A. , Jones Robert B. , Crotchfelt Kimberly , Moncada Jeanne , Jungkind D. , Turner Buffy , Peyton Cynthia , Kelly James F. , Weiss Judith B. , Rosenstraus Maurice 
Published: July 2002

ABSTRACT

The COBAS AMPLICOR CT/NG test for
Neisseria gonorrhoeae
cross-reacts with certain strains of nonpathogenic
Neisseria
species. In some strains, the target sequence is identical to that of
N. gonorrhoeae
, whereas other strains have a small number of mismatches within the regions recognized by the primers or probe used in the COBAS AMPLICOR NG test. These cross-reactive strains are occasionally present in urogenital specimens, causing false-positive results in the COBAS AMPLICOR NG test. Analysis of the data generated in a large multicenter clinical trial showed that 2.9% of the specimens gave signals between
A
660
s of 0.2 and 3.5 but that one-half of these equivocal specimens did not contain
N. gonorrhoeae
. Most of these equivocal specimens were correctly classified as true positive or true negative by retesting in duplicate and defining a PCR-positive result as two of three results with an
A
660
of ≥2.0. If specimens had been classified as positive or negative based on a single test result using a cutoff of an
A
660
of 0.2, specificity would have ranged from 96.2 to 98.9% depending on specimen type, sex, and presence of symptoms. By employing the equivocal zone-retesting algorithm, specificity increased to 98.6 to 99.9% with little effect (0.1 to 4.9% decrease) on sensitivity in most specimen types, enabling the test to achieve a positive predictive value of at least 90% in populations with a prevalence of 4% or higher. In lower-prevalence populations, the test could be used to screen for presumptive infections that would have to be confirmed by an independent test.
Francisella tularensis
Strain Typing Using Multiple-Locus, Variable-Number Tandem Repeat Analysis
Authors: Farlow Jason , Smith Kimothy L. , Wong Jane , Abrams Michelle , Lytle Michael , Keim Paul 
Published: July 2002

ABSTRACT

Francisella tularensis
, the etiological agent of tularemia, is found throughout the Northern hemisphere. After analyzing the
F. tularensis
genomic sequence for potential variable-number tandem repeats (VNTRs), we developed a multilocus VNTR analysis (MLVA) typing system for this pathogen. Variation was detected at six VNTR loci in a set of 56 isolates from California, Oklahoma, Arizona, and Oregon and the
F. tularensis
live vaccine strain. PCR assays revealed diversity at these loci with total allele numbers ranging from 2 to 20, and Nei's diversity index values ranging from 0.36 to 0.93. Cluster analysis identified two genetically distinct groups consistent with the current biovar classification system of
F. tularensis
. These findings suggest that these VNTR markers are useful for identifying
F. tularensis
isolates at this taxonomic level. In this study, biovar B isolates were less diverse than those in biovar A, possibly reflecting the history of tularemia in North America. Seven isolates from a recent epizootic in Maricopa County, Ariz., were identical at all VNTR marker loci. Their identity, even at a hypervariable VNTR locus, indicates a common source of infection. This demonstrates the applicability of MLVA for rapid characterization and identification of outbreak isolates. Future construction of reference databases will allow faster outbreak tracking as well as providing a foundation for deciphering global genetic relationships.
Molecular Fingerprinting of
Clostridium difficile
Isolates: Pulsed-Field Gel Electrophoresis versus Amplified Fragment Length Polymorphism
Authors: Klaassen Corné H. W. , van Haren Hanneke A. , Horrevorts Alfons M. 
Published: July 2002

ABSTRACT

Two molecular fingerprinting techniques, pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP), were used to investigate the epidemiological relatedness among
Clostridium difficile
isolates from suspected outbreaks in three general hospitals. Analysis by PFGE yielded inconclusive data as a result of extensive DNA degradation. Although this degradation could be prevented to a certain extent by the inclusion of thiourea in the electrophoresis buffer, the weak DNA banding patterns obtained in this way were still far from optimal. AFLP data were obtained by using fluorescently labeled PCR primers and analysis on an ABI PRISM automated DNA analysis platform. AFLP analysis yielded high resolution and highly reproducible DNA fingerprinting patterns from which the epidemiological relatedness among the isolates could easily be determined. AFLP results could be readily obtained within 24 h, whereas 3 to 4 days were routinely required to complete the lengthy PFGE protocol. AFLP clearly proved to be a much more fail-safe fingerprinting method for
C. difficile
isolates, especially for those isolates for which a standard PFGE procedure yielded inconclusive results due to DNA degradation.
Diagnosis of Human Leptospirosis by Monoclonal Antibody-Based Antigen Detection in Urine
Authors: Saengjaruk Patcharin , Chaicumpa Wanpen , Watt George , Bunyaraksyotin Gaysorn , Wuthiekanun Vanaporn , Tapchaisri Pramuan , Sittinont Chuanpit , Panaphut Thanachai , Tomanakan Kanchana , Sakolvaree Yuwaporn , Chongsa-Nguan Manas , Mahakunkijcharoen Yuvadee , Kalambaheti Thareerat , Naigowit Pimjai , Wambangco Michael Angelo L. , Kurazono Hisao , Hayashi Hideo 
Published: July 2002

ABSTRACT

Hybridomas secreting specific monoclonal antibodies (MAb) to all members of the genus
Leptospira
(clone LF9) and those that are specific only to the pathogenic species (clones LD5 and LE1) were produced. MAb LF9, which was immunoglobulin G1 (IgG1), reacted to a 38-kDa component of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated whole-cell lysates of all
Leptospira
spp., while MAb LD5 and MAb LE1, which were IgG1 and IgG2a, respectively, reacted to the 35- to 36-kDa components of all serogroups of the pathogenic species of
Leptospira.
The MAb LD5 was used in a dot blot-enzyme-linked immunosorbent assay (dot-ELISA) for detecting
Leptospira
antigen in urine samples serially collected from two groups of patients diagnosed with leptospirosis, i.e., 36 clinically diagnosed patients and 25
Leptospira
culture confirmed patients. Their serum samples were tested serologically by IgM Dipstick assay, indirect immunofluorescence assay (IFA), and/or microscopic agglutination test (MAT). Urine samples of 26 patients diagnosed with other illnesses and 120 healthy individuals served as controls. For the first group of patients, who had been ill for an average of 3.4 days before hospitalization, the IgM Dipstick test, IFA, and MAT were positive for 69.4, 70.0, and 85.7% of patients, while the
Leptospira
antigenuria tested by the MAb-based dot-ELISA was positive for 75.0, 88.9, 97.2, 97.2, and 100% of patients on days 1, 2, 3, 7, and 14 of hospitalization, respectively. All but 1 of 11 patients whose serum samples collected on the first day of hospitalization were IgM seronegative, were positive by urine antigen test on day 1. This is strong evidence that detection of antigen in urine can provide diagnostic information that could be useful in directing early therapeutic intervention. The MAT was positive in 10 of 12 patients (83.3%) of the 25 culture-positive
Leptospira
patients who had been ill for an average of 5.04 days before hospitalization, and the
Leptospira
antigen was found in 64.0, 84.0, 96.0, 100, 100, 100, and 100% of the patients' urine samples collected on days 1, 2, 3, 4, 5, 6, and 7 of hospitalization, respectively.
Leptospira
antigenuria was found in 3 of the 26 patients diagnosed with other illnesses and 1 of the 120 healthy controls. The reasons for this positivity are discussed. The detection of antigen in urine by the monoclonal antibody-based dot-ELISA has high potential for rapid, sensitive, and specific diagnosis of leptospirosis at a low cost.
Detection and Multigenic Characterization of a Herpesvirus Associated with Malignant Catarrhal Fever in White-Tailed Deer (
Odocoileus virginianus
) from Missouri
Authors: Kleiboeker Steven B. , Miller Margaret A. , Schommer Susan K. , Ramos-Vara Jose A. , Boucher Magalie , Turnquist Susan E. 
Published: July 2002

ABSTRACT

Between 1998 and 2001, tissues from four captive white-tailed deer were observed to have histologic lesions of systemic lymphocytic vasculitis. These lesions suggested malignant catarrhal fever, although epizootic hemorrhagic disease and bluetongue were included in the differential diagnosis. Initial diagnostic efforts, including virus isolation and reverse transcription-PCR for epizootic hemorrhagic disease virus and bluetongue virus, failed to identify an etiologic agent. However, consensus primer PCR targeted to the herpesvirus DNA polymerase gene detected viral genomic DNA in each of these four cases. Nucleotide sequence analysis of the amplified product demonstrated that the detected virus was identical over the compared region to the recently described malignant catarrhal fever virus of white-tailed deer (H. Li, N. Dyer, J. Keller, and T. B. Crawford, J. Clin. Microbiol. 38:1313-1318, 2000). Additional nucleotide sequencing of both the DNA polymerase gene and DNA packaging gene followed by phylogenetic analysis solidified this newly recognized herpesvirus as a member of the
Gammaherpesvirinae
and suggests that this virus, along with ovine herpesvirus 2, alcelaphine herpesvirus 1, alcelaphine herpesvirus 2 and caprine herpesvirus 2, may be part of a separate clade within this subfamily.
Application of the C
18
-Carboxypropylbetaine Specimen Processing Method to Recovery of
Mycobacterium avium
subsp.
paratuberculosis
from Ruminant Tissue Specimens
Authors: Thornton Charles G. , MacLellan Kerry M. , Stabel Judith R. , Carothers Christine , Whitlock Robert H. , Passen Selvin 
Published: July 2002

ABSTRACT

The causative agent of Johne's disease is
Mycobacterium avium
subsp.
paratuberculosis
. This is a chronic, debilitating gastrointestinal disorder that affects ruminants and is responsible for significant economic loss. The specimen processing method that combines C
18
-carboxypropylbetaine (CB-18) treatment and lytic enzyme decontamination has been shown to improve the diagnosis of mycobacterioses. This processing method was applied to the isolation of
M. avium
subsp.
paratuberculosis
from ruminant tissue samples. The BACTEC 12B liquid culture system was used but was supplemented with 1% egg yolk emulsion, 4 μg of mycobactin J, and 0.5% pyruvate (12B/EMP) for use in conjunction with this method. The final concentration of antibiotics used was 10 μg of vancomycin, 30 μg of amphotericin B, and 20 μg of nalidixic acid (VAN) per ml. A 7H10-based solid medium was also used that included mycobactin J, pyruvate, and VAN but excluded the egg yolk emulsion (7H10/MPV). Several
M. avium
subsp.
paratuberculosis
isolates were examined during the evaluation of this processing method. It was observed that treatment with lytic enzymes stimulated the growth of
M. avium
subsp.
paratuberculosis
; however, the growth of one isolate was markedly inhibited due to the presence of vancomycin. Subsequently, the vancomycin concentration in the VAN formulation was reduced to 2 μg/ml. A blinded panel of 60 previously characterized tissue samples from bovine and bison were then processed and analyzed by smear and culture. Historically, 31 and 37 specimens were classified as positive by histology and culture, respectively. The overall sensitivity and specificity of smear relative to culture following CB-18 processing were 97.6 and 89.5%, respectively. The 12B/EMP/VAN liquid culture system recovered
M. avium
subsp.
paratuberculosis
from 39 specimens, whereas 7H10/MPV and Herrold's egg yolk media recovered
M. avium
subsp.
paratuberculosis
from 26 and 16 specimens, respectively. The average times to positive were 7.4 ± 8.3, 29.9 ± 2.6, and 24 ± 0 days, respectively. The contamination rates were 4.8, 22.6, and 20.0%, respectively.
Evidence of the Presence of IS
1245
and IS
1311
or Closely Related Insertion Elements in Nontuberculous Mycobacteria outside of the
Mycobacterium avium
Complex
Authors: Keller Andreas P. , Beggs Marjorie L. , Amthor Beate , Bruns Fabian , Meissner Peter , Haas Walter H. 
Published: July 2002

ABSTRACT

A PCR assay based on the simultaneous detection of IS
1245
and IS
1311
was developed and used to determine the host range of these insertion elements. Specific PCR products were observed in
Mycobacterium malmoense
,
Mycobacterium scrofulaceum,
and
Mycobacterium nonchromogenicum
, indicating that IS
1245
and IS
1311
are not limited to the
Mycobacterium avium
complex.
Tetrazolium Microplate Assay as a Rapid and Inexpensive Colorimetric Method for Determination of Antibiotic Susceptibility of
Mycobacterium tuberculosis
Authors: Caviedes Luz , Delgado Jose , Gilman Robert H. 
Published: July 2002

ABSTRACT

The emergence of multidrug-resistant tuberculosis underscores the need for low-cost, rapid methods to determine the susceptibility of
Mycobacterium tuberculosis
to antibiotics. A new, rapid, easily read, and inexpensive colorimetric method with a tetrazolium indicator performs this determination as quickly and accurately as the more expensive Alamar Blue technique.