巢式聚合酶链反应检测麻风菌利福平耐药基因的研究

目的提高检测麻风菌的利福平耐药基因（rpoB）的敏感性。方法设立含聚合酶链反应增效剂（Q-solution）试验组和无PCR反应增效剂对照组，以比较增效剂对PCR的影响；采用巢式PCR，对第一轮PCR未扩增出rpoB基因390bp片段的标本，再进行第二轮PCR，扩增出rpoB突变集中区域的193bp片段；之后用双脱氧链终止法，对阳性PCR产物直接测序。结果Q-solution可显著提高PCR的敏感性，但是常规PCR检测rpoB基因的敏感性仅为45．2％。在确立巢式PCR最佳条件后，使用Q-solution可使其敏感性提高至90．5％。结论PCR反应增效剂和巢式PCR相结合可明显提高检测rpoB的敏感性和特异性。

Improved detection of Mycobacterium leprae rifampin resistance with nested-PCR

Objective To improve the sensitivity of PCR for studying the rifampin resistant gene (rpoB) of M.Leprae from clinical samples.Method After comparing the results of PCR with Q-Solution (PCR Enhancer) and without Q-solution,Nested-PCR was then applied for the detection of rpoB gene.Results Although PCR Enhancer improves the amplification of rpoB gene, the sensitivity of routine PCR is only 45. 2%.The sensitivity of Nested-PCR for detecting mutants in rpoB gene can further increase to 90.5% after selection of optimum parameters.Conclusion The combinaion of PCR Enhancer and Nested-PCR improves the sensitivity and specificity of PCR for detection of rifampin resistant gene of M. Leprae.