中文摘要：

目的评价利福平寡核苷酸探针杂交技术（RIFO杂交）和PCR-限制性片段长度多态性（PCR-RFLP）在结核分枝杆菌（MTB）耐利福平（RIF）和异烟肼（INH）快速检测中的应用价值。方法选取121株北京地区MTB菌株，分别采用RIFO杂交技术和PCR—RFLP检测RIF耐药相关基因rpoB核心区和INH耐药相关基因katG315位点突变，并对所有菌株的rpoB基因核心区进行测序验证。结果RIFO杂交检测发现，91，5％（65／71）的RIF耐药株和92．9％（52／56）的耐多药菌株（至少对RIF和INH耐药）存在rpoB基因核心区突变，而RIF敏感株中未发现突变；RIFO杂交与测序结果完全一致，测序结果中有突变的位点在RIFO杂交中均有相应的野生型杂交信号缺失；PCR-RFLP结果显示，INH耐药株中katG315突变率为60.6％（40／66）。结论rpoB基因核心区可作为RIF耐药检测的分子标志及耐多药的筛选指标；RIFO杂交技术是检测MTB耐RIF的快速、准确的实验方法，具有推广及潜在的临床应用价值；PCR—RFLP可检测出大部分INH耐药株，可作为临床INH耐药性检测的辅助手段。

英文摘要：

Objective To evaluate the rifampin oligonucleotide assay （RIFO assay ） and polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） in the rapid detection of rifampin （RIF） and isoniazid （INH） resistant Mycobacterium tuberculosis （MTB）. Methods Total 121 MTB strains isolated from Beijing, China were analyzed. The RIFO assay and PCR-RFLP were used to detect mutations in the RIF resistance associated gene rpoB hot-spot region and INH resistance associated gene katG locus 315, respectively. The rpoB hot-spot region of all the strains was sequenced for confirmation. Results Sixty-five （91.5%） of 71 RIF resistant and52 （92.9%） of 56 multi-drug resistant （resistant to at least RIF and INH） strains were found to harbor mutations in the rpoB hot-spot region by RIFO assay. No mutation was detected in RIF sensitive strains. The results of RIFO assay were concordant with that of sequencing, in which the corresponding wildhybridization signal of mutant allele was absent. katG315 substitution （AGC→ACC） was found in 40 （60.6%） of 66 INH resistant strains by PCR- RFLP. Condusions rpoB gene hot-spot region analysis can serve as a genetic marker for detection of RIF resistant and MDR isolates. The RIFO assay is a rapid, simple, and highly specific method for detection of RIF resistant strains, and can be a practical clinical tool in routine diagnosis. Most of INH resistant strains can be detected by PCR-RFLP, thus it can be used as a complementary method in clinical work.