NDLI: Direct detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction and DNA hybridization.

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Ability of clinical laboratories to detect antimicrobial agent-resistant enterococci.

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Direct detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction and DNA hybridization.

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Evidence of multiple taxa within commercially available reference strains of Corynebacterium xerosis.

Reproducibility of lysis-centrifugation cultures for quantification of Mycobacterium avium complex bacteremia.

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Identification of Actinobacillus actinomycetemcomitans by leukotoxin gene-specific hybridization and polymerase chain reaction assays.

Development of a competitive enzyme-linked immunosorbent assay for detection of bovine, ovine, porcine, and equine antibodies to vesicular stomatitis virus.

Detection of Toxoplasma gondii in venous blood from AIDS patients by polymerase chain reaction.

Development of a rapid and efficient restriction endonuclease analysis typing system for Clostridium difficile and correlation with other typing systems.

Curvilinear-gradient high-performance liquid chromatography for identification of mycobacteria.

Development and evaluation of a blood-free medium for determining growth curves and optimizing growth of Rochalimaea henselae.

Detection of adenovirus in clinical specimens by polymerase chain reaction and liquid-phase hybridization quantitated by time-resolved fluorometry.

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Recovery of Borrelia burgdorferi by filtration.

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Evaluation of the L-pyrrolidonyl-beta-naphthylamide hydrolysis test for the differentiation of members of the families Enterobacteriaceae and Vibrionaceae.

Analysis of serotype-specific antibodies to Trichosporon cutaneum types I and II in patients with summer-type hypersensitivity pneumonitis with monoclonal antibodies to serotype-related polysaccharide antigens.

N-acetyl-D-glucosamine medium improves recovery of Haemophilus influenzae from sputa of patients with cystic fibrosis.

Abolish Mycobacterium paratuberculosis strain 18.

Usefulness of DNA fingerprinting in combating tuberculosis.

In Vitro Susceptibility Testing of Haemophilus influenzae: Review of New National Committee for Clinical Laboratory Standards Recommendations

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Direct detection of Mycobacterium tuberculosis in sputum by polymerase chain reaction and DNA hybridization.

Content Provider	PubMed Central
Author	Nolte, F. S. Metchock, B. Mcgowan, J. E. Edwards, A. Okwumabua, O. Thurmond, C. Mitchell, P. S. Plikaytis, B. Shinnick, T.
Abstract	A polymerase chain reaction (PCR) assay for the rapid diagnosis of pulmonary tuberculosis was developed by using oligonucleotide primers to amplify a fragment of IS6110, an insertion sequence repeated multiple times in the chromosome of Mycobacterium tuberculosis. Sediment obtained from sputa processed by the N-acetyl-L-cysteine-NaOH method was suspended in a simple lysis buffer and was heated at 100 degrees C for 30 min prior to amplification. A dUTP-uracil N-glycosylase PCR protocol was used to prevent false-positive test results because of the carryover of products from previous amplification reactions. The 317-bp amplicon was detected by direct gel analysis and Southern blotting and then hybridization with a biotin-labeled internal probe. Hybrid molecules were detected by using a commercially available avidin-alkaline phosphatase-chemiluminescent substrate system (Tropix, Inc., Bedford, Mass.). The analytical sensitivity of the assay was 10 fg of purified mycobacterial DNA. The limits of detection by culture (Middlebrook 7H11 agar and Lowenstein-Jensen medium) and by PCR were equivalent in terminal dilution experiments for organism suspensions and positive sputa. An internal control was used to detect the presence of amplification inhibitors in each negative reaction mixture. DNA was purified from inhibitory specimens by phenol-chloroform extraction and ethanol precipitation. PCR results were compared with results of microscopy and conventional culture for the detection of M. tuberculosis in 313 sputum specimens. There were 124 specimens that were positive for M. tuberculosis by conventional methods and 113 (91%) that were positive by PCR. PCR detected 105 of 110 (95%) of the smear-positive and 8 of 14 (57%) of the smear-negative specimens. There were no false-positive results by PCR (specificity, 100%). This PCR assay innovations that make application of this new technology feasible in clinical microbiology laboratories.
Starting Page	1777
File Format	PDF
ISSN	1098660X
e-ISSN	1098660X
Journal	Journal of Clinical Microbiology
Issue Number	7
Volume Number	31
Language	English
Publisher Date	1993-07-01
Access Restriction	Open
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Microbiology (medical)

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