bluuebunny/crossref_metadata_2025_split huggingface dataset:
Purification and Characterization of a 43-Kilodalton Extracellular Serine Proteinase from
Cryptococcus neoformans
Authors: Yoo Jae il , Lee Yeong Seon , Song Chul-Yong , Kim Bong Su 
Published: February 2004

ABSTRACT

An extracellular proteinase was purified from culture filtrates of
Cryptococcus neoformans
NHPY24 by DEAE ion-exchange chromatography and gelatin affinity column chromatography with azoalbumin as the substrate. The molecular mass of the purified enzyme was 43 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, its pH optimum was 7.0 to 8.0, and maximal activity was obtained at pH 7.5 and 37°C. By isoelectric focusing, the purified enzyme had a pI of 4.77. Enzyme activity was inhibited by serine proteinase inhibitors such as phenylmethylsulfonyl fluoride and diisopropylfluorophosphate. The purified enzyme was thus a serine proteinase. It hydrolyzed natural substrates including hemoglobin, β-casein, and gamma globulin.
Viable but Nonculturable Bacteria Are Present in Mouse and Human Urine Specimens
Authors: Anderson Marc , Bollinger Daphne , Hagler Ashley , Hartwell Hadley , Rivers Bryan , Ward Kristie , Steck Todd R. 
Published: February 2004

ABSTRACT

The presence of viable but nonculturable bacteria in human clean-catch and mouse bladder-isolated urine specimens was investigated. Viable but nonculturable bacteria are alive but do not give rise to visible growth under nonselective growth conditions. Urine specimens obtained from human female volunteers with or without an active urinary tract infection were found to contain, on average, significantly more viable than culturable forms of bacteria. Additional support for the presence of viable but nonculturable cells in urine specimens considered sterile was obtained from examination of urine specimens obtained directly from the bladder of healthy mice. Because the viability assay used to study the viable but nonculturable condition is by necessity growth independent, and hence indirect, the accuracy of this assay that scores cells with intact cell membranes as being viable was studied. Greater than 95% of
Escherichia coli
cells exposed to lethal doses of UV irradiation were found to lose their membrane integrity within a day, a time frame similar to that used to examine urine specimens. These data suggest that viable but nonculturable cells can occur within regions of the urinary tract previously considered sterile.
Comparison of Sensitivities of Virus Isolation, Antigen Detection, and Nucleic Acid Amplification for Detection of Equine Influenza Virus
Authors: Quinlivan Michelle , Cullinane Ann , Nelly Maura , van Maanen Kees , Heldens Jacco , Arkins Sean 
Published: February 2004

ABSTRACT

Four seronegative foals aged 6 to 7 months were exposed to an aerosol of influenza strain A/Equi/2/Kildare/89 at 10
6
50% egg infective doses (EID
50
)/ml. Nasopharyngeal swabs were collected for 10 consecutive days after challenge. Virus isolation was performed in embryonated eggs, and the EID
50
was determined for all positive samples. The 50% tissue culture infective dose was determined using Madin-Darby canine kidney (MDCK) cells. Samples were also tested by an in vitro enzyme immunoassay test, Directigen Flu A, and by reverse transcription-PCR (RT-PCR) using nested primers from the nucleoprotein gene and a single set of primers from the matrix gene. RT-PCR using the matrix primers and virus isolation in embryonated eggs proved to be the most sensitive methods for the detection of virus. The Directigen Flu A test was the least sensitive method. The inclusion of 2% fetal calf serum in the viral transport medium inhibited the growth of virus from undiluted samples in MDCK cells but was essential for the maintenance of the virus titer in samples subjected to repeated freeze-thaw cycles.
Comparison of Mycosis IC/F and Plus Aerobic/F Media for Diagnosis of Fungemia by the Bactec 9240 System
Authors: Meyer Marie-Hélène , Letscher-Bru Valérie , Jaulhac Benoît , Waller Jocelyn , Candolfi Ermanno 
Published: February 2004

ABSTRACT

Fungemia is associated with a high mortality rate. We compared the performance of the Mycosis IC/F selective fungal medium and the Plus Aerobic/F standard bacteriological medium for the diagnosis of fungemia on the Bactec 9240 automatic system. We retrospectively analyzed 550 blood culture pairs composed of one Mycosis IC/F vial and one Plus Aerobic/F vial, drawn in 187 patients with fungemia. The positivity rate by vial was significantly higher on Mycosis IC/F medium than on Plus Aerobic/F medium (88.0% versus 74.9%,
P
< 0.0001). The positivity rate for fungus detection on Plus Aerobic/F medium fell to 26.9% when bacteria were present in the same vial. The positivity rate by patient was also significantly higher on Mycosis IC/F medium than on Plus Aerobic/F medium (92.5% versus 75.9%,
P
< 0.0001). A marked superiority of Mycosis IC/F medium was demonstrated for diagnosis of
Candida glabrata
fungemia (31 of 31, 100%, versus 18 of 31, 58.1%,
P
< 0.0001). The mean detection time was significantly shorter on Mycosis IC/F medium than on Plus Aerobic/F medium (28.9 ± 22.2 h versus 36.5 ± 24.6 h,
P
< 0.0001). The mean time saving was 8.8 h for
Candida albicans
and 43.7 h for
C. glabrata
. Mycosis IC/F medium enabled more sensitive and earlier diagnosis, particularly for the two strains most frequently responsible for fungemia,
C. albicans
and
C. glabrata
, and also in the event of the concomitant presence of both yeasts and bacteria. In patients with risk factors, it would thus appear to be sensible to draw a Mycosis IC/F vial in addition to the standard bacteriological vials.
Multicenter Evaluation of the VERSANT Hepatitis B Virus DNA 3.0 Assay
Authors: Yao Joseph D. C. , Beld Marcel G. H. M. , Oon Lynette Lin Ean , Sherlock Christopher H. , Germer Jeffrey , Menting Sandra , Se Thoe Su Yun , Merrick Linda , Ziermann Rainer , Surtihadi Johan , Hnatyszyn H. James 
Published: February 2004

ABSTRACT

The VERSANT hepatitis B virus (HBV) 3.0 Assay (branched DNA [bDNA]) (referred to herein as VERSANT 3.0) was evaluated at four external sites for analytical sensitivity, specificity, reproducibility, linearity of quantification, and limits of detection. In addition, each of the test evaluation sites provided HBV DNA-positive clinical samples that were previously analyzed by one of three commercially available HBV DNA quantitative tests: Digene Hybrid Capture II HBV DNA Test (Digene); VERSANT HBV DNA 1.0 Assay (bDNA) (VERSANT 1.0); and COBAS AMPLICOR HBV Monitor Test (COBAS AMPLICOR). These samples were reexamined using VERSANT 3.0. The results from these studies showed that VERSANT 3.0 has high specificity (99.3%), excellent reproducibility (between-run coefficient of variation [CV] = 1.6 to 9.4%; within-run CV = 6.5 to 20.7%), and a broad linear range of quantification (2.0 × 10
3
to 1.0 × 10
8
HBV DNA copies/ml) that facilitate the monitoring of HBV DNA levels at diagnosis and throughout the course of treatment. In general, correlation was very good between results obtained from clinical samples analyzed by VERSANT 3.0 and the comparative HBV DNA quantitative assays (VERSANT 1.0,
R
2
= 0.900; Digene,
R
2
= 0.985; COBAS AMPLICOR,
R
2
= 0.771). The greatest differences in comparative quantitation occurred at HBV DNA levels approaching the limits of the dynamic ranges for the comparative assays. The performance characteristics of the new VERSANT 3.0 assay demonstrated that it provides a reliable and robust method for routinely monitoring serum HBV DNA levels in assessing disease activity and determining response to antiviral treatment.
Molecular Characterization of Resistance to Mupirocin in Methicillin-Susceptible and -Resistant Isolates of
Staphylococcus aureus
from Nasal Samples
Authors: Chaves Fernando , García-Martínez Jesus , de Miguel Sonia , Otero Joaquín R. 
Published: February 2004

ABSTRACT

A total of 15 of 101 (14.8%) nasal methicillin-resistant
Staphylococcus aureus
(MRSA) isolates exhibited mupirocin resistance (Mup
r
) compared with 1 of 154 (0.6%) methicillin-susceptible
Staphylococcus aureus
isolates. A total of 14 (93%) isolates exhibiting high-level Mup
r
belonged to a single clone. Horizontal plasmid transfer and transmission of Mup
r
strains contribute to a high incidence of Mup
r
MRSA at our institution.
Comparison of
Acinetobacter baumannii
Isolates from United Kingdom Hospitals with Predominant Northern European Genotypes by Amplified-Fragment Length Polymorphism Analysis
Authors: Spence Richard P. , van der Reijden Tanny J. K. , Dijkshoorn Lenie , Towner Kevin J. 
Published: February 2004

ABSTRACT

Acinetobacter baumannii
isolates collected between 1999 and 2001 from 46 United Kingdom hospitals were compared with previously identified northern European genotypes by amplified-fragment length polymorphism (AFLP) analysis. Two predominant northern European genotypes associated with outbreaks in the mid-1980s had been superseded by new outbreak-associated genotypes.
Development of a New Serological Test for SerotypingHaemophilus parasuisIsolates and Determination of Their Prevalence in North America
Authors: Tadjine M. , Mittal K. R. , Bourdon S. , Gottschalk M. 
Published: February 2004

ABSTRACTHaemophilus parasuiscauses polyserositis in swine. Fifteen serovars have been characterized by immunodiffusion test, but many field strains are not typeable. Isolates (n= 300) ofH. parasuisfrom animals in North America were serotyped by a new indirect hemagglutination test. The test was rapid and effective for serotyping ofH. parasuis, and serovars 4, 5, 13, and 7 were the most prevalent serotypes.
Use of an Internal Positive Control in a Multiplex Reverse Transcription-PCR To Detect West Nile Virus RNA in Mosquito Pools
Authors: Eisler Diane L. , McNabb Alan , Jorgensen Danielle R. , Isaac-Renton Judith L. 
Published: February 2004

ABSTRACT

We report on the use of West Nile virus Armored RNA as an internal positive control (IPC) for the extraction and reverse transcription-PCR (RT-PCR) of RNA extracted from field-collected mosquitoes and on a multiplex real-time Taqman RT-PCR to simultaneously detect the 3′ noncoding region of West Nile virus and the West Nile virus NS5-2 region comprising the IPC. Mosquito pools from the province of British Columbia, Canada (
n
= 635), were tested in duplicate and found to be negative for West Nile virus and positive for the IPC. Known West Nile virus-positive supernatants from mosquito pools from the provinces of Alberta and Manitoba were tested in duplicate and found to be positive for both regions of the West Nile virus genome. The mean cycle threshold (Ct) value for the IPC in batch extraction controls ± 2 standard deviations was found to be 36.43 ± 1.78 cycles. IPCs of 98.4% (624) of West Nile virus-negative pools fell within this range, indicating the reproducibility of RNA extraction and RT-PCR for pools varying in mosquito genus and number. A comparison of mosquito pool genera revealed no significant genus effect on the Ct value of the IPC. The incorporation of West Nile virus Armored RNA as an IPC allows monitoring of RNA extraction and RT-PCR and detection of false-negative results due to failures in these processes or to PCR inhibition, respectively.
Identification of a New Type of
Babesia
Species in Wild Rats (
Bandicota indica
) in Chiang Mai Province, Thailand
Authors: Dantrakool Anchalee , Somboon Pradya , Hashimoto Tetsuo , Saito-Ito Atsuko 
Published: February 2004

ABSTRACT

A new type of rodent babesia, which resembled
Babesia microti
but was phylogenetically placed closest, with the highest level of statistical support, to
Babesia canis
, a canine babesia, was identified in Thai
Bandicota indica
in Thai provinces to which malaria is endemic. Close watch should be kept on human babesiosis in Thailand.
Resolution of Discrepant Results for
Candida
Species Identification by Using DNA Probes
Authors: Coignard Catherine , Hurst Steven F. , Benjamin Lynette E. , Brandt Mary E. , Warnock David W. , Morrison Christine J. 
Published: February 2004

ABSTRACT

Candida
species bloodstream isolates were collected from institutions participating in an active, population-based surveillance for candidemia. Species identifications were performed locally and then confirmed at the Centers for Disease Control and Prevention (CDC) by phenotype-based methods. Discrepancies in species identification between the referring institution and the CDC were noted for 43 of 935 isolates (4.6%). A DNA probe-based species identification system (PCR-enzyme immunoassay [EIA]) was then used to resolve these discrepancies. The PCR-EIA result was identical to the CDC phenotypic identification method for 98% of the isolates tested. The most frequently misidentified species was
Candida glabrata
(37% of all discrepant identifications). Such misidentifications could lead to the administration of inappropriate therapy given the propensity of
C. glabrata
to develop resistance to azole antifungal drugs.
Detection of the
Candida
Antigen Mannan in Cerebrospinal Fluid Specimens from Patients Suspected of Having
Candida
Meningitis
Authors: Verduyn Lunel Frans M. , Voss Andreas , Kuijper Ed J. , Gelinck L. B. S. , Hoogerbrugge Peter M. , Liem K. L. , Kullberg Bart Jan , Verweij Paul E. 
Published: February 2004

ABSTRACT

Cerebrospinal fluid samples from five patients from which
Candida
cells were cultured were tested for the presence of mannan. Samples from four patients categorized as having proven candidosis reacted positively. Samples from the remaining patient and from patients with other central nervous system infections were negative. Detection of mannan may be valuable in the diagnosis of
Candida
meningitis.
Direct Colorimetric Assay for Rapid Detection of Rifampin-Resistant
Mycobacterium tuberculosis
Authors: Abate Getahun , Aseffa Abraham , Selassie Alemayehu , Goshu Solomon , Fekade Bekele , WoldeMeskal Dawit , Miörner Håkan 
Published: February 2004

ABSTRACT

The colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was standardized for direct detection of rifampin-resistant
Mycobacterium tuberculosis
in sputum samples. The sensitivity and specificity of the direct MTT assay matched those of the standard indirect susceptibility assay on 7H10 medium, and interpretable results were obtained for 98.5% of the samples within 2 weeks. Traditional methods of in vitro drug susceptibility testing are time consuming and laborious. Susceptibility tests on clinical isolates require 6 to 9 weeks, and tests conducted directly on smear-positive samples take about 3 weeks (International Union Against Tuberculosis and Lung Disease, The public health service national tuberculosis reference laboratory and the national laboratory network. Minimum requirements, role and operation in a low-income country, Paris, France, 1998, and P. T. Kent and G. P. Kubica, Public health mycobacteriology. A guide for the level III laboratory, Centers for Disease Control and Prevention, Atlanta, Ga., 1985). More-rapid methods are available but are very expensive for routine use under program conditions in countries with high levels of tuberculosis endemicity.
Characterization of
Haemophilus segnis
, an Important Cause of Bacteremia, by 16S rRNA Gene Sequencing
Authors: Lau Susanna K. P. , Woo Patrick C. Y. , Mok Mo-yin , Teng Jade L. L. , Tam Victoria K. P. , Chan Katie K. H. , Yuen Kwok-yung 
Published: February 2004

ABSTRACT

We describe the application of 16S rRNA gene sequencing in defining eight cases of bacteremia due to
Haemophilus
species other than
Haemophilus influenzae
(non-
H. influenzae
bacteremia) during a 7-year period. The first case of acute pyelonephritis due to
Haemophilus segnis
is also reported. In contrast to the extremely rare incidence of
H. segnis
infections reported previously, our results suggested that
H. segnis
is an important cause of non-
H. influenzae
bacteremia.
Development, Implementation, and Impact of Acceptability Criteria for Serologic Tests for Infectious Diseases
Authors: Crump John A. , Corder Jennifer R. , Henshaw Nancy G. , Reller L. Barth 
Published: February 2004

ABSTRACT

Serologic testing is essential for the diagnosis of some infectious diseases and yet is fraught with potential pitfalls. All parts of the diagnostic process must be optimized to ensure that serologic tests perform adequately. Recognizing that a lack of clinical data and correctly timed, paired sera frequently led to uninterpretable serology results at our laboratory, we developed and implemented simple acceptability criteria for serologic tests. We assessed the impact of these criteria by comparing submissions and results for the year before and the year after implementation of the criteria. The number of serologic tests performed declined by 25% after implementation of the acceptability criteria, despite an increase in requests for serologic tests. Inappropriate testing of acute-phase sera alone fell from 49 to 0% (
P
< 0.001) for the tests monitored. Appropriate submission of paired sera rose from 9 to 19% (
P
= 0.006). The proportion of results classified as interpretable rose from 52 to 100% (
P
< 0.001). We recommend that acceptability criteria be developed and applied to samples submitted to clinical microbiology laboratories for serologic testing.
Evaluation of Commercial Enzyme-Linked Immunosorbent Assay Kits for Detection of Tuberculosis in Argentinean Population
Authors: Imaz María Susana , Comini Marcelo Alberto , Zerbini Elsa , Sequeira María Delfina , Latini Omar , Claus Juan Daniel , Singh Mahavir 
Published: February 2004

ABSTRACT
Pathozyme-Myco G (Myco G), M, A, and TB complex plus (Omega Diagnostics Ltd., Alloa, Scotland) were evaluated for the serological diagnosis of pulmonary tuberculosis (TB) in an Argentinean population. Sera from 58 patients with pulmonary TB, 24 subjects with pulmonary mycobacteriosis or mycoses (pulmonary MM group), and 45 subjects with other underlying disorders (control group) were analyzed. The sensitivities of the tests ranged from 29% (Myco M) to 82% (Myco G) in smear-positive patients (17 subjects) and from 29% (TB complex plus) to 49% (Myco G) in smear-negative patients (41 subjects). The specificities of the assays varied from 93% (Myco M) to 100% (Myco G and TB complex plus) in controls and from 62% (Myco A) to 96% (TB complex plus) in the pulmonary MM group. Overall, for the diagnosis of smear-negative patients, Myco G had the best characteristics, with a sensitivity of 49% and specificities of 100% for controls and 75% for the pulmonary MM group; after its combination with TB complex plus, its sensitivity improved to 59%. Nevertheless, despite its relatively poor capacity to discriminate between pulmonary TB and pulmonary MM, Myco G, alone or in combination with TB complex plus, would be a useful diagnostic tool for patients with suspected pulmonary TB living in areas where the relative prevalence of pulmonary MM was low.
Genotypic and Phenotypic Characterization of Drug-Resistant
Mycobacterium tuberculosis
Isolates from Rural Districts of the Western Cape Province of South Africa
Authors: Streicher E. M. , Warren R. M. , Kewley C. , Simpson J. , Rastogi N. , Sola C. , van der Spuy G. D. , van Helden P. D. , Victor T. C. 
Published: February 2004

ABSTRACT

Genotypic and phenotypic analysis of drug-resistant
Mycobacterium tuberculosis
isolates from the Western Cape Province of South Africa showed that drug resistance is widespread and recently transmitted. Multidrug-resistant (MDR) isolates comprise 40% of this collection, and a large pool of isoniazid monoresistance may be a future source of MDR tuberculosis.
Virulence Properties and Characteristics of Shiga Toxin-Producing
Escherichia coli
in São Paulo, Brazil, from 1976 through 1999
Authors: Vaz T. M. I. , Irino K. , Kato M. A. M. F. , Dias Â. M. G. , Gomes T. A. T. , Medeiros M. I. C. , Rocha M. M. M. , Guth B. E. C. 
Published: February 2004

ABSTRACT

Twenty-nine Shiga toxin-producing
Escherichia coli
(STEC) strains were identified in a collection of 2,607 isolates from patients with diarrhea in São Paulo, Brazil, from 1976 to 1999. The STEC strains belonged mainly to serotypes O111:HNM (HNM, nonmotile) (13 of 29 [44.8%]), O111:H8 (7 of 29 [24%]), and O26:H11 (4 of 29 [13.8%]);
stx
1
eae
(26 of 29 [89.6%]), in combination with either enterohemorrhagic
E. coli hlyA
(11 of 26 [42%]) or
astA
(24 of 26 [92.3%]), prevailed. The O111 STEC strains were distinguished by their inability to decarboxylate lysine. The predominance of STEC O111 and O26 since the late 1970s and the identification of STEC serotypes O55:H19, O93:H19, and O118:H16 in association with human infections in Brazil are described for the first time.
Evaluation of Sensititre Automated Reading and Incubation System for Automated Reading of Sensititre Broth Microdilution Susceptibility Plates
Authors: Chapin Kimberle C. , Musgnug Michael C. 
Published: February 2004

ABSTRACT
The Sensititre Automated Reading and Incubation System (ARIS) performs automated reading of Sensititre identification and susceptibility plates. ARIS susceptibility results were compared to manually read results. A total of 212 isolates (3,352 organism-antimicrobial agent combinations) yielded 99.4% essential agreement and categorical error rates of 0.2% minor, 0% major, and 1.0% very major. ARIS yields accurate susceptibility results compared to plates read manually.
Onychomycosis Due to
Emericella quadrilineata
Authors: Gugnani Harish C. , Vijayan Vanan K. , Tyagi Puneet , Sharma Sonal , Stchigel Alberto M. , Guarro Josep 
Published: February 2004

ABSTRACT

Nondermatophytic fungi are increasingly being reported as etiological agents of onychomycosis. We describe here a case of hand nail infection caused by
Emericella quadrilineata
(anamorph
Aspergillus tetrazonus
), a species not so far known to be an etiological agent of onychomycosis.
Enterococcus cecorum
Empyema Thoracis Successfully Treated with Cefotaxime
Authors: Woo Patrick C. Y. , Tam Dorothy M. W. , Lau Susanna K. P. , Fung Ami M. Y. , Yuen Kwok-Yung 
Published: February 2004

ABSTRACT

We report the first case of
Enterococcus cecorum
empyema thoracis and spontaneous bacterial peritonitis in a 44-year-old man with underlying cirrhosis. The patient responded to cefotaxime (MIC, 0.25 μg/ml) treatment and drainage of the empyema. Susceptibility of
E. cecorum
to expanded-spectrum cephalosporins could be due to its production of types of penicillin-binding proteins similar to those produced by
Streptococcus
species rather than to those produced by
Enterococcus
species (as predicted by phylogenetic analysis of the 16S rRNA gene sequences).
Culture-Negative Pericarditis Caused by
Neisseria meningitidis
Serogroup C
Authors: Moumile Kaoutar , Carbonnelle Etienne , Dessemme Philippe , Tamisier Daniel , Iserin Franck , Houdouin Véronique , Nassif Xavier , Berche Patrick 
Published: February 2004

ABSTRACT

We describe a case of primary purulent culture-negative pericarditis caused by
Neisseria meningitidis
serogroup C occurring in an 8-month-old previously healthy boy, which was detected in pericardial fluid by broad-spectrum PCR amplification.
Isolation of
Neosartorya pseudofischeri
from Blood: First Hint of Pulmonary Aspergillosis
Authors: Järv H. , Lehtmaa J. , Summerbell R. C. , Hoekstra E. S. , Samson R. A. , Naaber P. 
Published: February 2004

ABSTRACT

We report a case of invasive pulmonary aspergillosis caused by
Neosartorya pseudofischeri
S. W. Peterson [anamorph
Aspergillus thermomutatus
(Paden) S. W. Peterson]. The diagnosis was initially based on a positive blood culture for a strain isolated from a neutropenic patient by means of a BACTEC 9050 blood culture system. The final diagnosis was established based on X-ray and computer tomography scan results as well as the detection of
Aspergillus
antigen in the patient's serum.