Abstract:
:Nucleic acid sequence capture extraction was coupled with LightCycler PCR amplification and product detection using real-time fluorescence for rapid, definitive detection of Mycobacterium bovis in lymph node specimens from 38 cattle with bovine tuberculosis lesions. PCR amplification of sequence-captured DNA using both a conventional heating block thermocycler and a LightCycler thermocycler was compared with culture and histopathological analyses. Conventional PCR enabled detection of 26 of 28 culture-positive specimens (93%) in approximately 9 h, and the LightCycler PCR detected 20 of 28 culture-positive specimens (71%) in only 30 min. Specific confirmation of Mycobacterium tuberculosis complex DNA was achieved by LightCycler PCR amplification using Syb Green 1 and an M. tuberculosis complex-specific Cy5-labeled fluorescence resonance energy transfer probe. The system described here enabled rapid and specific laboratory confirmation of bovine tuberculosis, and this is the first report of the detection of M. bovis in tissues using LightCycler PCR. The fluorescence technology used in the study has potential to allow development of a high-throughput molecular diagnostic test for bovine tuberculosis.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Taylor MJ,Hughes MS,Skuce RA,Neill SDdoi
10.1128/JCM.39.4.1272-1278.2001subject
Has Abstractpub_date
2001-04-01 00:00:00pages
1272-8issue
4eissn
0095-1137issn
1098-660Xjournal_volume
39pub_type
杂志文章abstract::The capsular-like envelope of Legionella pneumophila strains Togus 1 (serotype 2) and Philadelphia 1 (serotype 1) was isolated and purified by column chromatography on Sepharose 6B. Antibody raised in rabbits to these two antigenic materials did not cross-react in gel diffusion. Upon electrophoresis followed by gel di...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.13.4.637-642.1981
更新日期:1981-04-01 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::A Swedish bovine and a Dutch human phage set for coagulase-negative staphylococci were used to phage type coagulase-negative staphylococci isolated from bovine milk from Minnesota dairy herds. A comparison was also made of the deoxyribonuclease activity of coagulase-negative staphylococci isolated from bovine milk in ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.10.3.299-301.1979
更新日期:1979-09-01 00:00:00
abstract::The RD2 region of the internalization-associated gene prtF1, which encodes the fibronectin-binding repeat domain type 2 of protein F1, plays a crucial role in the entry of group A streptococci (GAS) into epithelial cells. A molecular study of the variability of the RD2 region was carried out with 77 independent Italia...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.42.2.639-644.2004
更新日期:2004-02-01 00:00:00
abstract::Porphyromonas gingivalis is implicated in the etiology of chronic periodontitis. Genotyping studies suggest that genetic variability exists among P. gingivalis strains; however, the extent of variability remains unclear and regions of variability remain largely unidentified. To assess P. gingivalis strain diversity, w...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00569-09
更新日期:2009-10-01 00:00:00
abstract::The way in which the extensive use of highly active antiretroviral therapy (HAART) has influenced the incidence of visceral leishmaniasis (VL) among human immunodeficiency type 1 (HIV-1)-infected patients is not yet understood. The present study assessed whether the incidence of symptomatic VL in HIV-infected patients...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.3.762-767.2002
更新日期:2002-03-01 00:00:00
abstract::In this report, we present a case of rhino-orbital mucormycosis in a 57-year-old female with poorly controlled diabetes mellitus. The only mold cultured at 25°C, 37°C, and 40°C from a specimen of the nasal crust was identified phenotypically and independently using nuclear ribosomal DNA sequence data as Thamnostylum l...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.06611-11
更新日期:2012-04-01 00:00:00
abstract::An organism resembling Corynebacterium striatum was isolated from the blood of a patient with acute aortic valvular insufficiency and no history of valvular heart disease. At autopsy, histopathologic examination of the aortic valve revealed pleomorphic gram-positive bacilli and destruction of valvular tissue. Our isol...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.1.8-10.1990
更新日期:1990-01-01 00:00:00
abstract::We report a case of disseminated Scedosporium/Pseudallescheria infection due to Pseudallescheria boydii sensu stricto after lung transplantation in a patient with cystic fibrosis. Dissemination occurred under voriconazole. Despite surgery and combination therapy with voriconazole, caspofungin, and terbinafine, the pat...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01840-09
更新日期:2010-05-01 00:00:00
abstract::The in vitro susceptibilities of 66 molecularly identified strains of the Mucorales to eight antifungals (amphotericin B, terbinafine, itraconazole, posaconazole, voriconazole, caspofungin, micafungin, and 5-fluorocytosine) were tested. Molecular phylogeny was reconstructed based on the nuclear ribosomal large subunit...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.06133-11
更新日期:2012-01-01 00:00:00
abstract::Staphylococcal cassette chromosome mec (SCCmec) types and Panton-Valentine leukocidin (PVL) gene carriage were compared among suspected community-associated methicillin-resistant Staphylococcus aureus MRSA (CA-MRSA) and health care-associated MRSA (HA-MRSA) isolates. CA-MRSA isolates carried the SCCmec type IV complex...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01706-06
更新日期:2007-03-01 00:00:00
abstract::An enzyme-linked immunosorbent assay (ELISA) for the detection and quantitation of human immunoglobulin M (IgM), IgG, and IgA antibodies against Yersinia enterocolitica by using lipopolysaccharides as antigens is described. The results obtained with the lipopolysaccharide ELISA were compared with the results of the wh...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.14.1.6-14.1981
更新日期:1981-07-01 00:00:00
abstract::An optimized reverse transcription (RT)-PCR protocol for the intravitam detection of rabies virus genomic RNA was tested with clinical samples obtained from 28 patients suspected of having rabies, 9 of whom were confirmed to have had rabies by postmortem examination. RT-PCR using saliva combined with an immunofluoresc...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.4.1117-1121.1998
更新日期:1998-04-01 00:00:00
abstract::This report presents the case of a patient with a massive pyelonephritis and a urosepsis caused by Streptococcus pneumoniae. This case is unusual as the focus was distant from the respiratory tract, the usual primary site of infection caused by this organism. No other primary site of infection was documented. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.9.4383-4385.2004
更新日期:2004-09-01 00:00:00
abstract::A singleplex PCR assay using a single primer pair targeting the putative sugar transporter gene was developed here to distinguish Cryptococcus neoformans var. grubii, Cryptococcus neoformans var. neoformans, and Cryptococcus gattii according to the distinct size of the amplicon. The interspecies and intravarietal hybr...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00064-13
更新日期:2013-06-01 00:00:00
abstract::Helicobacter pylori strains possessing the cag pathogenicity island (PaI) are associated with the development of gastroduodenal diseases, including gastric cancer. cag PaI products induce the secretion of interleukin-8 (IL-8) from epithelial cells and facilitate the translocation of CagA into the cell cytosol. In East...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.2.786-790.2005
更新日期:2005-02-01 00:00:00
abstract::Concentrations of Mycobacterium avium-M. intracellulare ranging from 10(-1) to 10(3) CFU/ml were added to blood, placed in Isolator tubes, and held at room temperature for intervals ranging from 4 h to 56 days before being processed (centrifugation and culture on Middlebrook 7H10 agar). At all concentrations tested, M...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.6.1211-1214.1991
更新日期:1991-06-01 00:00:00
abstract::Evaluations of screening tests for bacteriuria have traditionally compared the test results with those of quantitative urine cultures. However, many patients with symptomatic urinary tract infections can have less than 10(5) CFU/ml in their urine. Therefore, the results of urine culture and three screening tests (Bac-...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.3.467-470.1987
更新日期:1987-03-01 00:00:00
abstract::The correlation between response to antiviral therapy and pretreatment viral load in patients with chronic hepatitis C has prompted the development of quantitative assays to measure viral load. The aim of our study was to assess the clinical relevance of the newly developed semiautomated PCR system COBAS HCV MONITOR v...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.7.2722-2725.2000
更新日期:2000-07-01 00:00:00
abstract::Approximately 8% of Rift Valley fever (RVF) cases develop severe disease, leading to hemorrhage, hepatitis, and/or encephalitis and resulting in up to 50% of deaths. A major obstacle in the management of RVF and other viral hemorrhagic fever cases in outbreaks that occur in rural settings is the inability to rapidly i...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01905-08
更新日期:2009-04-01 00:00:00
abstract::Quantitative PCR (qPCR) is more sensitive than microscopy for detecting Pneumocystis jirovecii in bronchoalveolar lavage (BAL) fluid. We therefore developed a qPCR assay and compared the results with those of a routine immunofluorescence assay (IFA) and clinical data. The assay included automated DNA extraction, ampli...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.06036-11
更新日期:2012-02-01 00:00:00
abstract::Many central vascular catheters (CVCs) are removed unnecessarily because current diagnostic methods for CVC-associated infection are unreliable. A quantitative PCR assay using primers and probe targeted to bacterial 16S ribosomal DNA was used to measure the levels of bacterial DNA in blood samples drawn through the CV...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.42.4.1402-1408.2004
更新日期:2004-04-01 00:00:00
abstract::Clonal spread and horizontal transfer in the spread of vancomycin resistance genes were investigated. Multiplex PCR, pulsed-field gel electrophoresis (PFGE), hybridization of enterococcal plasmids with the vanA and vanB probes, and sequencing of a fragment of vanB were used in the analysis. Before May 1996, 12 vancomy...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.12.3934-3939.1999
更新日期:1999-12-01 00:00:00
abstract::We determined the E-Test and National Committee for Clinical Laboratory Standards standardized agar dilution MICs of ceftazidime, ciprofloxacin, piperacillin, and tobramycin for Pseudomonas aeruginosa during tests of 100 rough and mucoid P. aeruginosa isolates from cystic fibrosis patients. The levels of agreement (+/...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.12.3191-3193.1995
更新日期:1995-12-01 00:00:00
abstract::A total of 525 cerebrospinal fluid (CSF) samples submitted during the 2007 and 2008 enteroviral seasons were included in a study to determine the prevalence of and potential risk factors for invalid Cepheid GeneXpert enterovirus assay (GXEA) results, as well as possible solutions for the problem. The invalid GXEA resu...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01238-09
更新日期:2009-09-01 00:00:00
abstract::The kinetics of delta antigen (HDAg) and anti-delta antibody (anti-HD) was analyzed in 22 acute delta hepatitis infections (11 coinfections and 11 superinfections), with an enzyme immunoassay developed by Organon Teknika and with two commercially available assays: Deltassay, a test for HDAg from Noctech and Abbott ant...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.7.1339-1342.1988
更新日期:1988-07-01 00:00:00
abstract::We compared examination of unstained and Gram-stained smears as methods of evaluating sputum quality. Of 100 slides examined, 96 (96%) were graded identically. Each discrepancy between methods was near the cutoff for specimen acceptability. We concluded that examination of unstained smears can provide an accurate, cos...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.24.1.143-144.1986
更新日期:1986-07-01 00:00:00
abstract::Naturally acquired murine cytomegalovirus (MCMV) infection in laboratory strains of mice induces antibody levels which are generally undetectable by standard techniques; therefore, MCMV has not been included routinely in mouse viral antibody screening programs. The relative sensitivity of three assay systems, the nucl...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.18.4.753-758.1983
更新日期:1983-10-01 00:00:00
abstract::A disk method for detecting hippurate hydrolysis by Campylobacter spp. was evaluated and compared with the conventional tube test used at the Centers for Disease Control (CDC) (G.K. Morris and C.M. Patton, p. 302-308, in E.H. Lennette, A. Balows, W.J. Hausler, Jr., and H.J. Shadomy, ed., Manual of Clinical Microbiolog...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.2.359-360.1989
更新日期:1989-02-01 00:00:00
abstract::To determine whether DNA detection of high-risk human papillomavirus could represent a reliable screening technique and a useful follow-up method, we investigated the performance of the Hybrid Capture II assay in detecting high-grade cervical lesions. The test positivity at 1.86 pg/ml had a high sensitivity (94.7%) an...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.42.1.366-368.2004
更新日期:2004-01-01 00:00:00