Abstract:
:By using a single synthetic oligonucleotide primer pair in the polymerase chain reaction, we amplified specific Shiga-like-toxin (SLT) gene segments from DNAs of 20 clinical Escherichia coli isolates, irrespective of whether they produce SLT-I, SLT-II, or heretofore uncategorized SLTs. These segments were not detectable in any of 20 nontoxigenic E. coli strains. The primers deduced from a conserved region among SLT genes are so-called degenerate-sequence primers; i.e., they contain intentionally introduced sequence ambiguities to overcome minor sequence variations within different SLT genes. In direct gel hybridization with genomic DNA, both primers recognized SLT-I and SLT-II DNA sequences. Amplified sequences of target DNA obtained by polymerase chain reaction were visualized after gel electrophoresis by ethidium bromide staining, and definitive identification of the amplification product as an SLT gene segment was achieved by hybridization to SLT-I- and SLT-II-specific 20-base oligonucleotide probes complementary to a portion of the amplified sequences but not to the primers. The detecting oligonucleotide probes shared only 30% base homology and were shown to recognize specifically SLT-I or SLT-II sequences within genomic DNA. Moreover, they were used to distinguish whether the amplified sequence originated from SLT-I or SLT-II genes. The PCR system with the primers described here is a powerful technique to amplify SLT sequences in E. coli strains that produce serologically distinct SLTs and will facilitate identification of these pathogens, particularly among a multitude of nonpathogenic E. coli strains.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Karch H,Meyer Tdoi
10.1128/JCM.27.12.2751-2757.1989subject
Has Abstractpub_date
1989-12-01 00:00:00pages
2751-7issue
12eissn
0095-1137issn
1098-660Xjournal_volume
27pub_type
杂志文章abstract::The biochemical characteristics and the presence of human Fc(gamma) receptors of 52 throat isolates of group C beta-hemolytic streptococci were examined. Among these isolates, 38 were identified as "Streptococcus milleri" and 14 were identified as "Streptococcus equisimilis." The differentiation of group C "S. equisim...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.24.5.705-707.1986
更新日期:1986-11-01 00:00:00
abstract::Ribotyping was compared with multilocus enzyme electrophoresis (MEE) for subtyping 305 Listeria monocytogenes isolates from clinical and nonclinical sources. For ribotyping, EcoRI-restricted genomic DNA fragments of L. monocytogenes strains were separated by agarose gel electrophoresis, and Southern blots were probed ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.32.12.2936-2943.1994
更新日期:1994-12-01 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::We compared the new Abbott TestPack (TP) respiratory syncytial virus (RSV) enzyme immunoassay (EIA) with cell culture and two commercial RSV EIAs (from Abbott Diagnostics and Kallestad Laboratories) by using split samples of fresh nasal washings from children with suspected RSV disease. Two tubes of HEp-2 cells were i...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.6.1395-1397.1990
更新日期:1990-06-01 00:00:00
abstract::An examination of 45 clinical isolates of Cryptococcus neoformans revealed an unusually high incidence of C. neoformans var. gattii in South Australia (65%) and in the Northern Territory (95%). In assessing all the available data from Australian isolates of C. neoformans, there appeared to be an endemic focus for the ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.2.430-431.1987
更新日期:1987-02-01 00:00:00
abstract::During a taxonomic investigation of Nocardia spp. and Streptomyces spp., we received 658 isolates from laboratories of both human and veterinary medicine. Our procedure leading to the identification of 92% of these isolates, the species that they represented, and a characteristic pattern of properties of the strains o...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.11.6.728-736.1980
更新日期:1980-06-01 00:00:00
abstract::While foreign-born persons constitute only 11% of the population in the state of Rhode Island, they account for more than 65% of incident tuberculosis (TB) annually. We investigated the molecular-epidemiological differences between foreign-born and U.S.-born TB patients to estimate the degree of recent transmission an...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01952-10
更新日期:2011-03-01 00:00:00
abstract::Ten strains of enterotoxigenic Escherichia coli producing heat-labile enterotoxin (LT) were preserved under 12 different conditions. After 1 month, 9 months, and 3 years of preservation, the cultures were recovered and examined for LT production. Preservation of the cultures on Dorset Egg Medium at 4 degrees C and pre...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.10.2326-2328.1991
更新日期:1991-10-01 00:00:00
abstract::Vaginal swab specimens may be preferable to cervical swab or urine specimens for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae because of the ease of specimen collection and transport. The purpose of this study was to evaluate whether vaginal swab specimens are equivalent to cervical swab specimens ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.8.3592-3596.2003
更新日期:2003-08-01 00:00:00
abstract::Serotyping of Chlamydia trachomatis strains usually requires three to six serial passages in shell vials to attain sufficient antigen for typing procedures. To circumvent this problem, we developed a rapid low-passage method for serotyping of C. trachomatis clinical isolates. Isolates with an inclusion count of greate...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.7.1333-1338.1991
更新日期:1991-07-01 00:00:00
abstract::Ectromelia virus, an orthopoxvirus that can cause extensive morbidity and mortality (mousepox) in colonized mice, has been epizootically responsible for serious disruption of biomedical research since 1930. The lack of a sensitive and specific serological assay for infection with this virus became apparent during outb...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.18.5.1220-1225.1983
更新日期:1983-11-01 00:00:00
abstract::Sapporo-like viruses (SLVs) are associated with acute gastroenteritis in humans. Due to a limited supply of available reagents for diagnosis, little is known about the incidence and pathogenicity of these viruses. We have developed a first-generation generic reverse transcriptase (RT) PCR assay based on a single prime...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.2.530-536.2000
更新日期:2000-02-01 00:00:00
abstract::The fluorescent amplified-fragment length polymorphism (FAFLP) assay potentially amplifies a unique set of genome fragments from each bacterial clone. It uses stringently hybridizing primers which carry a fluorescent label. Precise fragment sizing is achieved by the inclusion of an internal size standard in every lane...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.5.1274-1279.1999
更新日期:1999-05-01 00:00:00
abstract::We compared the relative levels of effectiveness of three commercial identification kits and three nucleic acid amplification tests for the identification of coryneform bacteria by testing 50 diverse isolates, including 12 well-characterized control strains and 38 organisms obtained from pediatric oncology patients at...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01849-07
更新日期:2008-03-01 00:00:00
abstract::A monoclonal antibody (MAb) immunoglobulin G2a (2125) was produced against a 60-kDa Legionella heat shock protein (HSP), recognizing a unique epitope common to all species of the genus Legionella. The antibody reacted in the immunoblot with 59 Legionella species and serogroups that were tested and showed no cross-reac...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.2.346-354.1991
更新日期:1991-02-01 00:00:00
abstract::Escherichia hermannii was isolated in pure culture from a patient with acute purulent conjunctivitis after a minor ocular injury. This is the first report of E. hermannii isolated as the sole pathogen from an infected site without prior antibiotic exposure, confirming the pathogenic potential of the microorganism. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01119-08
更新日期:2008-11-01 00:00:00
abstract::Monitoring antiretroviral therapy requires that human immunodeficiency virus type 1 (HIV-1) viremia assays are applicable to all distinct variants. This study evaluates the performance of three commercial viral load assays-Versant HIV-1 RNA bDNA v3.0, Cobas AmpliPrep/Cobas TaqMan HIV-1, and NucliSens HIV-1 EasyQ v1.2-...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02414-07
更新日期:2008-09-01 00:00:00
abstract::Since their first detection in 1988, penicillin-resistant Streptococcus pneumoniae isolates have rapidly spread in Iceland to account for close to 20% of all pneumococcal disease in that country by 1993. The major component (70%) of the resistant pneumococci identified from 1989 to 1992 was the progeny of a single mul...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.4.1375-1381.2000
更新日期:2000-04-01 00:00:00
abstract::Serum samples collected from 68 patients (age, mean +/- the standard deviation [SD], 56.3 +/- 12.8 years) at admission who were subsequently molecularly diagnosed as having hepatitis E and from 2,781 individuals who were assumed not to have been recently infected with hepatitis E virus (HEV; negative controls; 52.9 +/...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.1.49-56.2005
更新日期:2005-01-01 00:00:00
abstract::Between 1991 and 1995, an apparent high rate of Staphylococcus warneri bacteremias at the Royal Children's Hospital, Melbourne, Victoria, Australia, raised the possibility of a virulent nosocomial strain. In a retrospective review of 30 S. warneri bacteremias in children, organisms were viable and verified in 22 episo...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.8.2174-2177.1997
更新日期:1997-08-01 00:00:00
abstract::A procedure for detecting pneumococcal capsular polysaccharides in immune complexes is described. Separation of antigen from immune complexes is achieved by electrophoresis at 56 degrees C. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1978-08-01 00:00:00
abstract::Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease (JD) in cattle and may be associated with Crohn's disease (CD) in humans. It is the slowest growing of the cultivable mycobacteria, and culture from clinical, veterinary, food, or environmental specimens can take 4 months or even lon...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01492-10
更新日期:2011-05-01 00:00:00
abstract::Seventy-nine methicillin-resistant Staphylococcus aureus (MRSA) strains, isolated during 1980 to 1990, were classified as MRSA Aggl- (14 strains) and MRSA Aggl+ (65 strains) strains on the basis of test results in slide agglutination assays designed to detect fibrinogen-binding protein (clumping factor) and protein A ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.32.1.143-147.1994
更新日期:1994-01-01 00:00:00
abstract::The PLEX-ID system is a novel technology that couples PCR amplification and electrospray ionization-mass spectrometry to identify pathogens directly in clinical specimens. The analytical performance of the PLEX-ID Broad Fungal assay was compared with that of traditional culture identification by using 91 characterized...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03282-12
更新日期:2013-06-01 00:00:00
abstract::Quantitative microdilution plate hybridization was used to identify 22 Mycobacterium species. DNAs of clinical strains were rapidly extracted and labeled with photoreactive biotin. Labeled DNAs were distributed into wells of a microdilution plate in which reference DNAs had been immobilized. After 2 h of hybridization...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.8.1596-1603.1991
更新日期:1991-08-01 00:00:00
abstract::In the last decade, the global emergence of carbapenem resistance in Enterobacteriaceae has posed great concern to public health. Data concerning the role of environmental contamination in the dissemination of carbapenem-resistant Enterobacteriaceae (CRE) are currently lacking. Here, we aimed to examine the extent of ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01992-12
更新日期:2013-01-01 00:00:00
abstract::Establishment of latently infected cell lines from primary effusion lymphomas (PEL) presently is the most efficient system for the propagation of clinical strains of human herpesvirus 8 (HHV-8) in culture. Here we describe a new approach to culture productively replicating HHV-8 from patient samples. A BJAB-derived B-...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.6.2866-2875.2005
更新日期:2005-06-01 00:00:00
abstract::In this work, we describe the assembly of a synthetic gene coding for several antigenic determinants found in different Leishmania infantum antigens. Selected epitopes were derived from the ribosomal proteins LiP2a, LiP2b, and LiP0 and from the histone H2A. The resulting gene was overexpressed in Escherichia coli eith...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.1.58-63.1998
更新日期:1998-01-01 00:00:00
abstract::The clinical performance of the cobas human papillomavirus (HPV) test for detection of high-grade disease in a colposcopy-referred population was compared with that of Hybrid Capture 2 (HC2). The overall agreement between the tests was 92.3%. Clinical sensitivity and specificity for detection of cervical intraepitheli...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01949-13
更新日期:2013-10-01 00:00:00
abstract::The number of reported cases of infections with non-serogroup O1 Vibrio cholerae in the United States has increased recently. These cases have almost invariably been associated with travel, seawater exposure, or the ingestion of shellfish. We report a case of bacteremic cellulitis caused by non-O1 V. cholerae that was...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.12.2874-2876.1989
更新日期:1989-12-01 00:00:00