Abstract:
:We describe a simple method for the rapid detection of bovine viral diarrhea virus (BVDV) that uses a one-tube reverse transcription PCR (RT-PCR) and total RNA extracted directly from a variety of bovine specimens, including whole blood and tissues. Reagents for both RT and PCR were combined in a one-tube, single-buffer system, and amplification was performed with a single uninterrupted thermal cycling program. Using the novel cationic surfactant tetradecyltrimethylammonium oxalate (Catrimox-14), we consistently extracted RT-PCR-quality RNA from specimens containing blood. Amplification with primers derived from conserved sequences within the BVDV 5'-untranslated region yielded a 244-bp product. Assay specificity was confirmed by ethidium bromide-stained gel electrophoresis and by chemiluminescence-assayed Southern blot hybridizations involving BVDV 5'-untranslated region-specific digoxigenin-labelled cDNA probes. The assay detection level was 0.1 50% tissue culture infectious dose of BVDV when ethidium bromide-stained gel electrophoresis was used and 0.01 50% tissue culture infectious dose of BVDV when Southern blot hybridization was used. Our method is an alternative to the conventional cell culture assays used in a diagnostic laboratory and is an improvement over existing RT-PCR assays for BVDV.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Hamel AL,Wasylyshen MD,Nayar GPdoi
10.1128/JCM.33.2.287-291.1995subject
Has Abstractpub_date
1995-02-01 00:00:00pages
287-91issue
2eissn
0095-1137issn
1098-660Xjournal_volume
33pub_type
杂志文章abstract::Identification of members of the Mycobacterium tuberculosis complex and the M. avium-M. intracellulare complex (MAC) directly from primary BACTEC cultures was evaluated by using acridinium-ester-labeled DNA probes (AccuProbe; GenProbe, Inc., San Diego, Calif.). In preliminary experiments, blood present in samples was ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.9.2427-2431.1992
更新日期:1992-09-01 00:00:00
abstract::A point prevalence culture survey was carried out to investigate the prevalence of fecal carriage of vancomycin-resistant enterococci (VRE) among patients admitted to an 800-bed general hospital where no VRE had been isolated previously. Twenty-two of 636 patients (3.5%) were found to be VRE carriers. Eighteen strains...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.11.2842-2846.1995
更新日期:1995-11-01 00:00:00
abstract::Carbapenem-resistant Acinetobacter baumannii isolates were obtained from 24 patients between March and May 2004 at the Centre Hospitalier de Polynésie Française, Tahiti, French Polynesia. The isolates were multidrug resistant, produced the carbapenemase OXA-23, and belonged to a single clone presenting several subtype...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.9.4826-4829.2005
更新日期:2005-09-01 00:00:00
abstract::A 42-plex clustered regularly interspaced short palindromic repeat (CRISPR)-based typing technique (spoligotyping) was recently developed at the French National Reference Center for Legionella. It allows the subtyping of the Legionella pneumophila sequence type 1/Paris pulsotype. In this report, we present the transfe...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00219-14
更新日期:2014-07-01 00:00:00
abstract::Diagnostic tests for respiratory viral infections have traditionally been performed on nasopharyngeal swabs or washings. Reverse transcriptase PCR (RT-PCR) is rapid, sensitive, and specific for viral infection diagnosis but is rarely applied to sputum samples. Thus, we evaluated the diagnostic yield of RT-PCR for dete...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.05841-11
更新日期:2012-01-01 00:00:00
abstract::In a clinical trial, Strep-A-Chek (a 10-min chromogenic test) was compared with the bacitracin disk susceptibility test for accuracy and turnaround time in the presumptive identification of Streptococcus pyogenes. Among 461 isolates of beta-hemolytic streptococci (344 throat isolates and 117 isolates from other sites)...
journal_title:Journal of clinical microbiology
pub_type: 临床试验,杂志文章
doi:10.1128/JCM.24.3.431-434.1986
更新日期:1986-09-01 00:00:00
abstract::The BACTEC NR660 blood culture system, which uses infrared spectroscopy to detect carbon dioxide generated by bacterial growth, was compared with the new medium formulation of the Oxoid Signal system. Two trials were conducted: a comparative study of 88 organisms in simulated blood cultures and a clinical trial of 3,3...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.7.1586-1590.1990
更新日期:1990-07-01 00:00:00
abstract::Noroviruses are the leading cause of outbreaks of gastroenteritis in the world. At present, norovirus genogroup II, genotype 4 (GII/4), strains are the most prevalent in many countries. In this study we investigated 55 outbreaks and 35 sporadic cases of norovirus-associated gastroenteritis in food handlers in food-cat...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01516-07
更新日期:2007-12-01 00:00:00
abstract::This study shows that an unbiased amplification method applied to equine arteritis virus RNA significantly improves the sensitivity of the real-time reverse transcription-quantitative PCR (RT-qPCR) recommended by the World Organization for Animal Health. Twelve viral RNAs amplified using this method were hybridized on...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01935-14
更新日期:2015-01-01 00:00:00
abstract::Seven rotavirus strains dually reactive to serotype G1- and G2-specific monoclonal antibodies (MAbs) from hospitalized children with rotavirus diarrhea were culture adapted. Six strains were neutralized with G1 antiserum to a higher titer than that of G2, and one was neutralized with G1 and G2 antisera to equal titers...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.11.5267-5269.2003
更新日期:2003-11-01 00:00:00
abstract::The NucliSENS easyMAG automated system was compared to the column-based Qiagen method for Epstein-Barr virus (EBV) or cytomegalovirus (CMV) DNA extraction from whole blood before viral load determination using the corresponding R-gene amplification kits. Both extraction techniques exhibited a total agreement of 81.3% ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01497-09
更新日期:2009-11-01 00:00:00
abstract::Cowdria ruminantium causes severe, often fatal disease in domestic ruminants, whereas wildlife species usually are not affected. Blood and bone marrow samples from healthy, free-ranging Zimbabwean ungulates were taken during translocation from areas harboring Amblyomma ticks and tested for the presence of C. ruminanti...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.9.2501-2504.1995
更新日期:1995-09-01 00:00:00
abstract::In the present study, we compared typed F. nucleatum strains with isolates from various human periodontal diseases, canines, and a Macaca mulatta monkey. All isolates displayed biochemical reactions similar to those of the typed strains in the API 20A system (Analytab Products, Plainville, N.Y.). The human and monkey ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.17.4.631-635.1983
更新日期:1983-04-01 00:00:00
abstract::A rapid dimethyl sulfoxide modification of an acid-fast technique was applied to direct fecal smears to monitor cryptosporidiosis in nonhuman primates. Brilliantly stained pink oocysts against a pale green background demonstrated well-preserved internal morphology and facilitated rapid, simple, noninvasive diagnosis w...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.19.6.952-953.1984
更新日期:1984-06-01 00:00:00
abstract::A previously described microbroth kinetic system (J. Meletiadis, J. F. Meis, J. W. Mouton, and P. E. Verweij, J. Clin. Microbiol. 39:478-484, 2001) based on continuous monitoring of changes in the optical density of fungal growth was used to describe turbidimetric growth curves of different filamentous fungi in the pr...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.10.4718-4725.2003
更新日期:2003-10-01 00:00:00
abstract::Clinical isolates of group B streptococci from body fluids and mucosal surfaces were examined for production of a trypsin-resistant antigen known as R protein. R protein was extracted with 1% trypsin from cells grown in a semidefined medium. The extracts were tested by immunodiffusion in agarose with a panel of antise...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.5.1050-1054.1989
更新日期:1989-05-01 00:00:00
abstract::Coprological examination based on egg detection in stool samples is currently used as the gold standard for the diagnosis of human fascioliasis. However, this method is not effective during the acute phase of the disease and has poor sensitivity during the chronic phase. Serodiagnosis has become an excellent alternati...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02875-13
更新日期:2014-03-01 00:00:00
abstract::In this study, we determined the sequences of four intimin variant genes detected in attaching and effacing Escherichia coli isolates of human origin. Three of them were novel and were designated eae-eta (eta), eae-iota (iota), and eae-kappa (kappa). The fourth was identical to the recently described eae-zeta (zeta), ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.12.4486-4492.2002
更新日期:2002-12-01 00:00:00
abstract::A large pool of refined endotoxin was prepared from Escherichia coli O113 by extraction with hot acqueous phenol. It was characterized chemically and biologically and will be available for a reference standard designated as reference endotoxin EC. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1976-01-01 00:00:00
abstract::Longitudinal data on the E6/E7 mRNA-based Aptima human papillomavirus (AHPV) assay exceeding three years in comparison to the gold standard Digene Hybrid Capture 2 (HC2) test are not available. We previously reported the cross-sectional data of the German AHPV Screening Trial (GAST) in which 10,040 women were recruite...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01177-18
更新日期:2019-01-02 00:00:00
abstract::The sequences of mink astroviruses collected from 11 farms in Denmark and Sweden were analyzed and found to be homologous with one another but different from those of other astroviruses. A species-specific reverse transcriptase-PCR for mink astrovirus was established and shown to be suitable for the analysis of clinic...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.11.5192-5194.2003
更新日期:2003-11-01 00:00:00
abstract::The performance of a disk diffusion test using broth from positive blood cultures as inoculum (direct disk diffusion [dDD]) was evaluated for a collection of 20 challenge isolates of Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa Isolates seeded into human blood were inoculated into Bactec Plu...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01678-17
更新日期:2018-02-22 00:00:00
abstract::Salmonella enterica subsp. enterica serovar Agona was not frequently encountered in Finland until an increase in rates of isolation among animal and feed was seen in 1994. A small outbreak among cattle farms in the regions of Oulu and Vaasa in northwestern Finland in 1994-1995 included eight farms. After the outbreak,...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.10.3648-3653.2002
更新日期:2002-10-01 00:00:00
abstract::Haemophilus influenzae type e biotype 4 was isolated from a single antemortem blood culture obtained from a 60-year-old, white male with abdominal carcinomatosis. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1978-01-01 00:00:00
abstract::An enzyme-linked immunosorbent assay method was used to classify bovine serum as positive, negative, or doubtful for antibodies to Brucella abortus. Spectrophotometric data from assays of 64 serologically positive and 32 serologically negative bovine sera were analyzed statistically to define the range of spectrophoto...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.11.4.398-401.1980
更新日期:1980-04-01 00:00:00
abstract::"Campylobacter pyloridis" has been recently described as a gastritis-associated bacterium. We studied 20 strains. The bacteria had most of the characteristics of Campylobacter spp. strains. They were hippurate negative and tolerant to triphenyl tetrazolium chloride (at 0.4 and 1 mg/ml). They grew on all the media comm...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.22.6.1007-1010.1985
更新日期:1985-12-01 00:00:00
abstract::Accurate and internationally comparable human papillomavirus (HPV) DNA genotyping is essential both for evaluation of HPV vaccines and for effective monitoring and implementation of vaccination programs. The World Health Organization (WHO) HPV Laboratory Network (LabNet) regularly issues international proficiency stud...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00840-12
更新日期:2012-07-01 00:00:00
abstract::Initially, multilocus enzyme electrophoresis was used to examine genetic relationships among 63 isolates of Mycobacterium bovis and 13 other members of the M. tuberculosis complex. The isolates were divided into five electrophoretic types, with a mean genetic diversity of 0.1. The strains were genetically homogenous, ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.5.1136-1142.1996
更新日期:1996-05-01 00:00:00
abstract::Campylobacter jejuni (20 strains) and Campylobacter coli (12 strains) were assigned to four biovars for each species based on phenotypic tests that were easy to perform and interpret. The resulting biotyping schemes offer a greater degree of distinction among C. jejuni and C. coli strains than any of the other biotypi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.20.5.990-992.1984
更新日期:1984-11-01 00:00:00
abstract::DNA colony hybridization with three 18- to 20-base-long synthetic oligonucleotide probes for cholera toxin (CT) was used to screen 12 clinical isolates of Aeromonas hydrophila. Under stringent hybridizing (overnight at 40 degrees C) and washing (1 h at 50 degrees C) conditions, nine strains reacted with the 32P-labele...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.1.57-61.1988
更新日期:1988-01-01 00:00:00