摘要
目的应用焦磷酸测序技术检测结核分枝杆菌rpoB基因突变,并探讨其在结核分枝杆菌利福平耐药性检测中的应用价值。方法利用焦磷酸测序技术,测定150株结核分枝杆菌临床分离株rpoB基因81个碱基的利福平抗药性决定区(RRDR),分析rpoB基因突变特点,并与绝对浓度法和MIC法药敏结果进行比较。结果150株结核分枝杆菌中66株耐药84株敏感,54株为耐多药菌株,rpoB基因突变涉及6个密码子12种突变基因型,最常见的突变位点及突变形式分别为531位TCG突变为TTG,占60.6%,526位CAC突变为TAC、GAC,占22.7%。耐药性检测结果表明,焦磷酸检测法与绝对浓度法二者的符合率为92.7%,与MIC法的符合率为97.8%。结论焦磷酸测序检测技术不仅是一种快速、准确、高通量的利福平分子药敏检测方法,同时也是结核分枝杆菌耐药性研究的有用工具。
Objective To detect the mutations of rpoB gene in Mycobacterium tuberculosis by pyrosequencing and to evaluate the values on detection of fifampin resistance in clinical isolates. Methods Using the new technology of pyrosequencing, the mutations in the fifampin resistance determining region (RRDR) of rpoB gene were analyzed. The results were compared with those obtained from methods of the absolute concentration and the minimum inhibitory concentration(M/C). Results Among the 150 Mycobacterium tuberculosis clinical isolates,84 were susceptible and 66 resistant to RIF. 54 of the 66 resistant isolates were muhidrug-resistant (MDR) strains. Ser531Leu and His526Asp or Tyr, including twelve different genotypes and six codons,were the most common mutations. In the drug susceptibility testing, the accordance rates of the pyrosequencing and the absolute concentration method as well as MIC were 92.7% and 97.8% respectively. Conclusion Not only is the pyrosequencing technology a fast, sensitive and high throughput method in detecting rifampin resistance in Mycobacterium tuberculosis, but also a useful tool in the research of rifampin resistantce mechanism.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2008年第2期77-80,共4页
Chinese Journal of Preventive Medicine
基金
上海市科委重点专项课题(05DZ22320)