Detection of Rifampin-Resistant Mycobacterium tuberculosis in Sputa by Nested PCR-Linked Single-Strand Conformation Polymorphism and DNA Sequencing
| Autor: | Sang Jae Kim, Eun Mi Park, Keun-Hwa Lee, Yoon-Hoh Kook, Gil-Han Bai, Seo-Jeong Kim, Bum Joon Kim, Bo-Na Park, Young Gil Park |
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| Rok vydání: | 2001 |
| Předmět: |
Microbiology (medical)
Polymerase Chain Reaction DNA sequencing law.invention Mycobacterium tuberculosis chemistry.chemical_compound law polycyclic compounds Humans Tuberculosis Antibiotics Antitubercular Polymorphism Single-Stranded Conformational Polymerase chain reaction biology Sputum Drug Resistance Microbial Mycobacteriology and Aerobic Actinomycetes Single-strand conformation polymorphism DNA-Directed RNA Polymerases Sequence Analysis DNA bacterial infections and mycoses biology.organism_classification rpoB Virology Molecular biology chemistry Nontuberculous mycobacteria Rifampin Nested polymerase chain reaction DNA |
| Zdroj: | Journal of Clinical Microbiology. 39:2610-2617 |
| ISSN: | 1098-660X 0095-1137 |
| Popis: | Either PCR-mediated single strand conformation polymorphism (SSCP) analysis or DNA sequencing of rpoB DNA (157 bp) can be used as a rapid screening method for the detection of mutations related to the rifampin resistance of Mycobacterium tuberculosis . However, due to the nonspecific amplification of rpoB DNA from nontuberculous mycobacteria these methods cannot be directly applied to clinical specimens such as sputa. We developed a nested PCR method that can specifically amplify the rpoB DNA of M. tuberculosis on the basis of rpoB DNA sequences of 44 mycobacteria. Nested PCR-linked SSCP analysis and the DNA sequencing method were applied directly in order to detect M. tuberculosis and determine its rifampin susceptibility in 56 sputa. The results obtained by nested PCR-SSCP and DNA sequencing were concordant with those of conventional drug susceptibility testing and DNA sequencing performed with culture isolates. |
| Databáze: | OpenAIRE |
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