Title: Clinical analysis of rpoB mutations of Mycobacterium tuberculosis by the PCR-SSCP
Abstract:Objective To establish PCR SSCP for analyzing rpoB mutation of Mycobacterium tuberculosis and to evaluate its use in clinics. Methods A rpoB fragment was amplified from M. tuberculosis strains and spu...Objective To establish PCR SSCP for analyzing rpoB mutation of Mycobacterium tuberculosis and to evaluate its use in clinics. Methods A rpoB fragment was amplified from M. tuberculosis strains and sputum specimens by PCR with a pair of primers designed according to the lifampin resistant determinant of rpoB. The fragments of rpoB were analyzed by single stranded conformation polymorphism (SSCP) and some fragments were sequenced. Results A 230 bp fragments of rpoB were amplified from all M. tuberculosis strains and 113 of 135 (83.7%) sputum samples that were positive for acid fast stain, but not from 16 species of nontuberculosis bacteria and sputum from nontuberculosis patients. In SSCP analysis, 130 of 140 (92.9%) strains resistant to lifampin in L J drug susceptibility test were rpoB mutants and 67 of 72 (93.1%) strains susceptible to lifampin were wild type. In sequencing analysis, 57 mutant fragments in SSCP were all found to have sequence changes and 2 of 10 wild types of fragments in SSCP to have sequence changes. Among the lifampin resistant strains, the multi drug resistant (MDR) strains were 76.4%. The MDR strains,92.5%(99/107) rpoB mutation in SSCP. Conclusion This PCR SSCP is an accurate and stable method to identify rpoB showed mutation of M. tuberculosis , and it is useful for quickly identifying the lifampin resistant or the MDR strains in clinics.Read More
Publication Year: 2002
Publication Date: 2002-01-01
Language: en
Type: article
Access and Citation
AI Researcher Chatbot
Get quick answers to your questions about the article from our AI researcher chatbot