Direct, automated detection of rifampin-resistant Mycobacterium tuberculosis by polymerase chain reaction and single-strand conformation polymorphism analysis
Détails
ID Serval
serval:BIB_A05CA2BA8748
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Direct, automated detection of rifampin-resistant Mycobacterium tuberculosis by polymerase chain reaction and single-strand conformation polymorphism analysis
Périodique
Antimicrobial Agents and Chemotherapy
ISSN
0066-4804 (Print)
Statut éditorial
Publié
Date de publication
10/1993
Volume
37
Numéro
10
Pages
2054-8
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Oct
Research Support, Non-U.S. Gov't --- Old month value: Oct
Résumé
A rapid screening test was recently established for the detection of mutations in the rpoB gene of Mycobacterium tuberculosis, a region identified as the locus for rifampin resistance (Rifr). The detection method involved the amplification by polymerase chain reaction (PCR) of the Rifr region and the identification of mutations by single-strand DNA conformation polymorphism analysis (SSCP) of the amplification products. Experience using two different PCR-SSCP formats for the evaluation of BACTEC cultures and sputum is presented here; the previously described manual procedure for the detection for the detection of radiolabelled amplification products and an automated SSCP by which fluorescein-labelled products were detected on a Pharmacia DNA sequencer apparatus. All 17 different Rifr mutations known to date were consistently detected. PCR-SSCP could be used for the evaluation of minimally grown cultures (BACTEC 12B medium with a growth index of < or = 100) and for direct screening of microscopically positive sputa with greater than 10 organisms per field (magnification, x250). Implementation of this technique could result in rapid detection of rifampin resistance in M. tuberculosis, a marker of multidrug-resistant tuberculosis.
Mots-clé
Base Sequence
DNA, Single-Stranded/*analysis
Drug Resistance, Microbial
Evaluation Studies
Humans
Molecular Sequence Data
Mutation/*genetics
Mycobacterium tuberculosis/*drug effects/*genetics
Nucleic Acid Conformation
Polymerase Chain Reaction/*methods
Polymorphism, Genetic
Rifampin/*pharmacology
Sputum/microbiology
Pubmed
Web of science
Création de la notice
25/01/2008 15:45
Dernière modification de la notice
20/08/2019 16:06