Abstract:
:A set of universal degenerate primers which amplified, by PCR, a 600-bp oligomer encoding a portion of the 60-kDa heat shock protein (HSP60) of both Staphylococcus aureus and Staphylococcus epidermidis were developed. However, when used as a DNA probe, the 600-bp PCR product generated from S. epidermidis failed to cross-hybridize under high-stringency conditions with the genomic DNA of S. aureus and vice versa. To investigate whether species-specific sequences might exist within the highly conserved HSP60 genes among different staphylococci, digoxigenin-labelled HSP60 probes generated by the degenerate HSP60 primers were prepared from the six most commonly isolated Staphylococcus species (S. aureus 8325-4, S. epidermidis 9759, S. haemolyticus ATCC 29970, S. schleiferi ATCC 43808, S. saprophyticus KL122, and S. lugdunensis CRSN 850412). These probes were used for dot blot hybridization with genomic DNA of 58 reference and clinical isolates of Staphylococcus and non-Staphylococcus species. These six Staphylococcus species HSP60 probes correctly identified the entire set of staphylococcal isolates. The species specificity of these HSP60 probes was further demonstrated by dot blot hybridization with PCR-amplified DNA from mixed cultures of different Staphylococcus species and by the partial DNA sequences of these probes. In addition, sequence homology searches of the NCBI BLAST databases with these partial HSP60 DNA sequences yielded the highest matching scores for both S. epidermidis and S. aureus with the corresponding species-specified probes. Finally, the HSP60 degenerate primers were shown to amplify an anticipated 600-bp PCR product from all 29 Staphylococcus species and from all but 2 of 30 other microbial species, including various gram-positive and gram-negative bacteria, mycobacteria, and fungi. These preliminary data suggest the presence of species-specific sequence variation within the highly conserved HSP60 genes of staphylococci. Further work is required to determine whether these degenerate HSP60 primers may be exploited for species-specific microbic identification and phylogenetic investigation of staphylococci and perhaps other microorganisms in general.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Goh SH,Potter S,Wood JO,Hemmingsen SM,Reynolds RP,Chow AWdoi
10.1128/JCM.34.4.818-823.1996subject
Has Abstractpub_date
1996-04-01 00:00:00pages
818-23issue
4eissn
0095-1137issn
1098-660Xjournal_volume
34pub_type
杂志文章abstract::Statistical analysis of a limiting dilution assay (LDA) showed that the occurrence of infectious human immunodeficiency virus type 1 (HIV-1)-harboring cells in serially diluted samples of peripheral blood mononuclear cells (PBMCs) of HIV-1-seropositive patients fits the model describing a single-hit Poisson distributi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.10.2535-2538.1992
更新日期:1992-10-01 00:00:00
abstract::The human metapneumovirus (hMPV) was recently identified and linked to acute respiratory tract infections (ARTI). To assess the clinical importance of this virus in infants and children, we developed a rapid and efficient reverse transcription-PCR-based screening method for a large volume of samples and tested retrosp...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.11.5520-5525.2005
更新日期:2005-11-01 00:00:00
abstract::Adolescent children were surveyed for colonization with Mycoplasma hominis and Ureaplasma urealyticum by culturing urine specimens. Rates were compared between three study groups: (i) 397 children attending parochial schools, (ii) 293 children attending an adolescent clinic specializing in adjustment problems, and (ii...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1975-09-01 00:00:00
abstract::Clostridium difficile is now recognized as the major agent responsible for nosocomial diarrhea in adults. Among the genotyping methods available, arbitrarily primed PCR (AP-PCR), PCR-ribotyping, and pulsed-field gel electrophoresis (PFGE) have been widely used for investigating outbreaks of C. difficile infections. Ho...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.7.2484-2487.2000
更新日期:2000-07-01 00:00:00
abstract::A new anti-human immunodeficiency virus type 1 and 2 (anti-HIV 1 and 2) test is described. It uses recombinant p24 and peptides covering gp32, gp41, and gp120 to identify HIV-1 and HIV-2 infections. This test has been shown to be specific (99.5%) and sensitive (99.8%). In this respect, the assay was equal or superior ...
journal_title:Journal of clinical microbiology
pub_type: 临床试验,杂志文章,多中心研究
doi:10.1128/JCM.32.2.403-406.1994
更新日期:1994-02-01 00:00:00
abstract::We studied the use of dried spots of bodily fluids (plasma, whole blood, and mother's milk) on filter paper as a means of sample collection and storage for human immunodeficiency virus type 1 (HIV-1) viral load testing under stringent field conditions. Plasma placed directly in lysis buffer, which is customarily used ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01919-06
更新日期:2007-03-01 00:00:00
abstract::The performances of a commercially available qualitative plasma PCR assay (AMPLICOR CMV test; Roche Diagnostics) and the pp65 antigenemia assay (AG) were evaluated for the monitoring of cytomegalovirus (CMV) viremia in 43 allogeneic stem cell transplant recipients. In addition, the suitabilities of both assays for tri...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.11.3938-3941.2001
更新日期:2001-11-01 00:00:00
abstract::The 37-kDa protein (P37) of Borrelia burgdorferi is an antigen that elicits an early immunoglobulin M (IgM) antibody response in Lyme disease patients. The P37 gene was cloned from a B. burgdorferi genomic library by screening with antibody from a Lyme disease patient who had developed a prominent humoral response to ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.3.548-552.1999
更新日期:1999-03-01 00:00:00
abstract::At the Islet Isolation Laboratory of the Scottish National Blood Transfusion Service, manual sterility testing data show that contamination rates are 57.7% for pancreas transport fluid, 4.3% for postpurification islet samples, and 0% for pretransplant islet samples. This pilot study presents the BacT/Alert System as a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00447-14
更新日期:2014-10-01 00:00:00
abstract::Clostridium difficile is the most common cause of health care-associated diarrhea. Accurate and rapid diagnosis is essential to improve patient outcome and prevent disease spread. We compared our two-step diagnostic algorithm, an enzyme immunoassay for glutamate dehydrogenase (GDH) followed by the cytotoxin neutraliza...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01648-10
更新日期:2010-12-01 00:00:00
abstract::Following intranasal administration of live influenza A virus vaccine or parenteral inoculation of inactivated influenza virus vaccine, immunoglobulin antibody to the influenza virus hemagglutinin was detected in nasal wash specimens from adult volunteers. Several observations supported the suggestion that this immuno...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.3.559-562.1987
更新日期:1987-03-01 00:00:00
abstract::Legionella is the causative agent for Legionnaires' disease (LD) and is responsible for several large outbreaks in the world. More than 90% of LD cases are caused by Legionella pneumophila, and studies on the origin and transmission routes of this pathogen rely on adequate molecular characterization of isolates. Curre...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01369-15
更新日期:2015-10-01 00:00:00
abstract::Classical microbiological diagnosis of human brucellosis is time-consuming, hazardous, and subject to variable interpretation. Intact-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was evaluated for the routine identification of Brucella spp. Analysis of mass peak patt...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01720-13
更新日期:2013-09-01 00:00:00
abstract::The frequencies of 10 opportunistic DNA viruses were determined by multiplex real-time PCR in paired cerebrospinal fluid (CSF) and brain tissue of HIV-infected individuals. In the CSF, viruses were detectable in 45/55 cases: JC virus (JCV) in 62%, Epstein-Barr virus (EBV) in 44%, cytomegalovirus (CMV) in 25%, varicell...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03321-12
更新日期:2013-03-01 00:00:00
abstract::The first World Health Organization (WHO) international standards (ISs) for nucleic acid amplification techniques were established two decades ago, with the initial focus on blood screening for three major viral targets, i.e., hepatitis C virus, hepatitis B virus, and human immunodeficiency virus 1. These reference ma...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,评审
doi:10.1128/JCM.01056-18
更新日期:2019-01-02 00:00:00
abstract::We report the recovery of Histoplasma capsulatum from blood specimens cultured for Mycobacterium sp. in BACTEC 13A radiometric medium. H. capsulatum was recovered from six of eight blood specimens submitted for mycobacterial cultures from five human immunodeficiency virus-positive individuals. Initial positive metabol...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.1.237-239.1992
更新日期:1992-01-01 00:00:00
abstract::Chlamydia pecorum is a significant pathogen of domestic livestock and wildlife. We have developed a C. pecorum-specific multilocus sequence analysis (MLSA) scheme to examine the genetic diversity of and relationships between Australian sheep, cattle, and koala isolates. An MLSA of seven concatenated housekeeping gene ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00992-13
更新日期:2013-08-01 00:00:00
abstract::The IS200-like insertion sequence (IS) is a 708-bp element recently found in Yersinia pestis. Its nucleotide sequence is 85% identical to that of IS200 recovered in most Salmonella enterica isolates. It is also present in multiple copies in Y. pseudotuberculosis. In contrast, this IS is found in some (biotype 1B strai...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.9.2231-2235.1996
更新日期:1996-09-01 00:00:00
abstract::Use of whole-genome sequencing (WGS) for routine mycobacterial species identification and drug susceptibility testing (DST) is becoming a reality. We compared the performances of WGS and standard laboratory workflows prospectively, by parallel processing at a major mycobacterial reference service over the course of 1 ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01480-17
更新日期:2018-01-24 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.11.3285-3290.1998
更新日期:1998-11-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.4.964-970.1999
更新日期:1999-04-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.9.2381-2385.1997
更新日期:1997-09-01 00:00:00
abstract::Ninety-one isolates of nutritionally variant streptococci (NVS) that were previously isolated from the human mouth were regarded as consisting of 7 Streptococcus defectivus isolates, 78 Streptococcus adjacens isolates, and 6 Gemella morbillorum isolates. However, recent references to the taxonomic reclassification of ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.8.2553-2556.1999
更新日期:1999-08-01 00:00:00
abstract::Bacteriuria due to Salmonella typhi usually occurs following recent typhoid fever or in chronic carrier states. Data from 18 patients with S. typhi bacteriuria, seen during 5 years, were analyzed. Fourteen patients had localized urinary tract infection due to S. typhi. Four others had bacteriuria, probably associated ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.7.1791-1792.1995
更新日期:1995-07-01 00:00:00
abstract::Several biochemical test systems were studied for their potential usefulness for the examination of strains of Campylobacter species. Most (81%) of the C. jejuni strains hydrolyzed sodium hippurate, but strains of C. fetus, C. sputorum, and C. fecalis did not. Some (46%) of the C. jejuni strains and all of the C. sput...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.15.6.1065-1073.1982
更新日期:1982-06-01 00:00:00
abstract::We describe a multiplex PCR assay to identify and discriminate between isolates of Campylobacter coli, Campylobacter jejuni, Campylobacter lari, and Campylobacter upsaliensis. The C. jejuni isolate F38011 lpxA gene, encoding a UDP-N-acetylglucosamine acyltransferase, was identified by sequence analysis of an expressio...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.12.5549-5557.2004
更新日期:2004-12-01 00:00:00
abstract::A Campylobacter-like organism was isolated from the ilea of normal hamsters. The organism was isolated from an ileal homogenate which was passed through a filter (0.65-micron pore size) and cultured on blood-agar plates in a microaerophilic atmosphere at 37 degrees C. Pinpoint translucent colonies were first observed ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.11.2497-2501.1989
更新日期:1989-11-01 00:00:00
abstract::A total of 50 strains of Staphylococcus aureus, including 41 methicillin-resistant S. aureus (MRSA) strains, were characterized by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins. The protein patterns contained 40-50 discrete bands and were highly reproducible. Partial ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.11.2574-2581.1989
更新日期:1989-11-01 00:00:00
abstract::We systematically evaluated 5 methods for testing daptomycin versus 48 Enterococcus faecalis, 51 Enterococcus faecium, and 50 Staphylococcus aureus isolates using (i and ii) broth microdilution (BMD) with 50-mg/liter calcium medium supplementation (reference method) and 30-mg/liter calcium medium supplementation (BMD3...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03439-13
更新日期:2014-06-01 00:00:00
abstract::Rapid tests for glucose, maltose, lactose, sucrose, and starch fermentation were performed on 112 strains of anaerobic bacteria. The tests were incubated under aerobic conditions, and results were read within 4 h. An overall correlation of 89% was achieved between the rapid tests and the Virginia Polytechnic Institut...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1976-03-01 00:00:00