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探讨反向线性杂交技术(reverse line blot assay,RLB)在结核分枝杆菌利福平(RFP)耐药性快速检测中的应用价值。采用RLB将包含rpoB基因核心区的扩增产物与标记特异性探针的膜进行杂交,共对121株结核分枝杆菌进行RFP耐药性检测,并将结果与常规药敏实验进行比较。结果发现,121株中有71株对RFP耐药,56株为多重耐药株;采用RLB共检测到65株RFP耐药株rpoB基因核心区存在突变,其灵敏度为91.5%(65/71);50株RFP敏感株中均未检测到突变,则特异性为100%(51/51);56株多耐药菌株中,92.9%(52/56)存在rpoB基因核心区突变。因此,用RLB检测结核分枝杆菌RFP耐药株具有快速,高效,特异性和灵敏度高的优点,且可用于多耐药菌株的筛选,具有推广和潜在的临床应用价值。
To investigate the value of reverse line blot assay (RLB) in the rapid detection of Mycobacterium tuberculosis rifampicin (RFP) resistance. A total of 121 strains of Mycobacterium tuberculosis were tested for RFP resistance by using RLB to hybridize the amplified product containing the core region of rpoB gene with the membrane of the labeled specific probe. The results were compared with conventional susceptibility testing. The results showed that 71 out of 121 strains were resistant to RFP and 56 were multidrug resistant strains. There were mutations in the core region of rpoB gene of 65 RFP resistant strains detected by RLB, with a sensitivity of 91.5% (65/71) ; No mutation was detected in 50 RFP-sensitive strains, and the specificity was 100% (51/51). Of the 56 multi-drug resistant strains, 92.9% (52/56) had mutations in the rpoB gene core region. Therefore, the detection of Mycobacterium tuberculosis RFP resistant strains with RLB has the advantages of rapid, efficient, specific and high sensitivity, and can be used for screening multi-drug resistant strains, which has the promotion and potential clinical application value.