Abstract:
:The detection of amplified products resulting from polymerase chain reactions (PCRs) remains a complicated process. To simplify the detection procedures, we developed a colorimetric microtiter plate hybridization assay for the specific detection of 5'-biotinylated PCR fragments of Mycobacterium leprae DNA. For this assay, an M. leprae DNA capture probe was made and immobilized on the wells of a microtiter plate. Hybridization of the biotin-labeled PCR fragments was detected through enzymatic color development. The resulting optical densities showed a logarithm-linear relationship with the amount of template DNA and corresponded to the intensity of the bands obtained through gel analysis and Southern blotting of the PCR products. The sensitivity of the assay was found to be 125 fg of genomic M. leprae DNA, or 20 lysed bacilli, revealing a detection limit similar to that of agarose gel analysis. The efficient coamplification of human DNA was used as a positive control for the presence of inhibitory substances in clinical material. For detection of human PCR products, a human DNA capture probe was also constructed for the colorimetric assay. This dual setup for hybridization, which thus detected both M. leprae and human DNA PCR products, was useful for ascertaining the presence of inhibiting substances in clinical specimens. All biopsy specimens (n = 10) from untreated patients with leprosy were positive. Apparently, this assay is more sensitive than microscopy, because biopsy specimens from half of the patients were negative upon histopathological examination. Biopsy specimens from three treated patients were negative, as were those from the three patients who did not have leprosy. We conclude that this colorimetric assay can replace agarose gel analysis and Southern hybridization, because it is as sensitive as those methods. Its advantages over conventional gel analysis and Southern hybridization are that it is less cumbersome and more rapid.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
van der Vliet GM,Hermans CJ,Klatser PRdoi
10.1128/JCM.31.3.665-670.1993subject
Has Abstractpub_date
1993-03-01 00:00:00pages
665-70issue
3eissn
0095-1137issn
1098-660Xjournal_volume
31pub_type
杂志文章abstract::When the standard procedure for determining antibiotic susceptibility of bacteria is used, the results are delayed, especially for bacteria that grow slowly, such as Helicobacter pylori. Treatment for this bacterium may involve clarithromycin, a compound for which resistance has been associated with point mutations on...
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
abstract::Collecting whole blood on filter paper simplifies the processing, transport, and storage of specimens used for the diagnosis of human immunodeficiency virus type 1 (HIV-1) and other tests. Specimens may be collected in tropical or rural areas with minimal facilities for handling specimens. To compare simulated tropica...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00359-08
更新日期:2008-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:2009-05-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.7.1940-1942.1993
更新日期:1993-07-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02364-05
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.9.2175-2179.1996
更新日期:1996-09-01 00:00:00
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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
abstract::Among 234 isolates comprising 26 different Candida species colonizing the oropharynx of 181 (54.3% of 399 surveyed) HIV-infected outpatients, 27 (11.7%) were fluconazole resistant. Antibacterial treatment was associated with increased rates of yeast colonization, while antiretroviral therapy and pneumococcal vaccinati...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00500-10
更新日期:2010-07-01 00:00:00
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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
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.4.1399-1403.2003
更新日期:2003-04-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.5.1134-1139.1992
更新日期:1992-05-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.8.3712-3718.2003
更新日期:2003-08-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00306-14
更新日期:2014-07-01 00:00:00
abstract::Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (O...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00304-18
更新日期:2018-06-25 00:00:00
abstract::Brucellosis is a globally significant zoonosis, the control of which is difficult and resource intensive. Serological tests form a vital part of a multifactorial approach to control and are often performed in large numbers. The aim of the present study was to develop a new assay to improve the efficiency, ease, and ef...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00919-09
更新日期:2009-10-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.12.2904-2908.1996
更新日期:1996-12-01 00:00:00
abstract::While outbreaks of animal anthrax zoonoses still regularly occur in France, little is known about the epidemiology links between them. We have used the eight-locus multilocus variable-number tandem repeat analysis typing technique against a collection of 50 Bacillus anthracis isolates from France. There were eight dis...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.12.4732-4734.2002
更新日期:2002-12-01 00:00:00
abstract::An outbreak of pseudobacteremia due to Streptococcus pyogenes (group A streptococci [GAS]) and methicillin-susceptible Staphylococcus aureus (MSSA) was traced to the venting procedure for aerobic bottles prior to their loading into the incubator of the BacT/Alert analyzer (Organon Teknika). Bacteria shed by a laborato...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.1.392-393.2001
更新日期:2001-01-01 00:00:00
abstract::The fortuitous finding that Aeromonas spp. grew well on Butzler Campylobacter selective medium (BCSA) in a microaerobic atmosphere at 42 degrees C prompted us to evaluate the performance of BCSA for selective isolation of aeromonads in comparison with ampicillin (30 micrograms/ml) sheep blood agar (ASBA30). Overall re...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.2.346-347.1989
更新日期:1989-02-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.2.467-469.1998
更新日期:1998-02-01 00:00:00
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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::Forty-four nonreplicate clinical isolates of Pseudomonas aeruginosa that were resistant to extended-spectrum cephalosporins (ceftazidime and cefepime) and aztreonam, that putatively produced an acquired extended- spectrum beta-lactamase (ESBL), according to the results of a double-disk synergy test, and that had been ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.6.2523-2529.2004
更新日期:2004-06-01 00:00:00
abstract::We present a duplex, real-time PCR assay for detection of Klebsiella pneumoniae carbapenemase (blaKPC) and New Delhi metallo-β-lactamase (blaNDM) genes. Accuracy was assessed with 158 Gram-negative bacillary isolates, including 134 carbapenemase producers. The assay had 100% sensitivity and specificity compared with r...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03062-12
更新日期:2013-04-01 00:00:00
abstract::The Triage parasite panel (BIOSITE Diagnostics, San Diego, Calif.) is a new qualitative enzyme immunoassay (EIA) panel for the detection of Giardia lamblia, Entamoeba histolytica/E. dispar, and Cryptosporidium parvum in fresh or fresh, frozen, unfixed human fecal specimens. By using specific antibodies, antigens speci...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.9.3337-3340.2000
更新日期:2000-09-01 00:00:00
abstract::Serial cerebrospinal fluid samples were obtained from 14 multiple sclerosis patients over a period of several months and were tested under code for Inoue-Melnick virus. In five of the positive patients, virus was present in 28 of 32 specimens collected over a period of 2 to 5 months. Four patients, from whom a total o...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.22.4.651-653.1985
更新日期:1985-10-01 00:00:00
abstract::The BACTEC MGIT 960 system, a fully automated, nonradiometric, noninvasive system for detection and drug susceptibility testing of mycobacteria, was evaluated for the ability to test susceptibilities to second-line drugs. In this study, which was carried out in three phases (phase I, mostly susceptible strains; phase ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
doi:10.1128/JCM.44.3.688-692.2006
更新日期:2006-03-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.44.5.1881-1883.2006
更新日期:2006-05-01 00:00:00
abstract::In the last 2 decades, a variety of different molecular typing methods have been developed to differentiate strains of Mycobacterium avium subsp. paratuberculosis. The most successful techniques are based on insertion sequences, repetitive loci, comparative genomics, or single nucleotide polymorphisms. In the present ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02031-07
更新日期:2008-04-01 00:00:00
abstract::We describe the adoption of nucleic acid amplification tests (NAAT) for Clostridium difficile diagnosis and their impact on stool rejection policies and C. difficile positivity rates. Of the laboratories with complete surveys, 51 (43%) reported using NAAT in 2011. Laboratories using NAAT had stricter rejection policie...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02177-13
更新日期:2014-02-01 00:00:00
abstract::Staphylococcus lugdunensis is a coagulase-negative staphylococcus that has several similarities to Staphylococcus aureus. S. lugdunensis is increasingly being recognized as a cause of prosthetic joint infection (PJI). The goal of the present retrospective cohort study was to determine the laboratory and clinical chara...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01769-09
更新日期:2010-05-01 00:00:00