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PCR-探针熔解分析法检测结核分枝杆菌利福平耐药性的临床应用
Authors
张灵霞
李国利
李庆阁
王倩
Publication date
10 September 2012
Publisher
Abstract
目的评价PCR-探针熔解分析法(probe melting analysis assay,PMAA)检测结核分枝杆菌(Mycobacterium tuberculosis,MTB)对利福平(rifampin,RFP)耐药性的临床应用价值。方法采用常规比例法和PCR-PMAA对512株MTB临床分离株进行RFP药敏试验,并进行RFP耐药相关基因rpoB DNA测序。结果以常规比例法为标准,PCR-PMAA检测RFP耐药性的敏感性、特异性分别为97.5%、96.2%,阳性和阴性预测值分别为94.1%、98.4%、符合率及约登指数分别为96.7%及0.93。193株比例法和PCR-PMAA两法检测均耐药的菌株,经DNA测序证实174株rpoB基因RFP耐药决定区(RFP resistance determining region,RRDR)发生单位点密码子突变,19株RRDR发生不同组合型的双位点密码子联合突变;5株比例法检出耐药、PCR-PMAA敏感的菌株,DNA测序rpoB基因未见突变;12株比例法检出敏感、PCR-PMAA耐药的菌株,DNA测序证实其中7株511位点密码子突变,5株533位点密码子突变;302株两法检测均敏感菌株DNA测序结果未见rpoB耐药相关的基因位点突变。PCR-PMAA检测MTB临床分离株rpoB基因突变结果与DNA测序结果完全一致。结论 PCR-PMAA法能高效检出MTB对RFP耐药的相关基因突变,用于检测MTB对RFP耐药性的敏感性高、特异性强,方法简便、快速、价廉,具有良好的临床应用前景
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Last time updated on 16/06/2016