Park Young Kil, Shin Sonya, Ryu Sungweon, Cho Sang Nae, Koh Won-Jung, Kwon O Jung, Shim Young Soo, Lew Woo Jin, Bai Gill Han
Korean Institute of Tuberculosis, Seoul 137-900, South Korea.
J Microbiol Methods. 2005 Nov;63(2):165-72. doi: 10.1016/j.mimet.2005.03.002.
The frequency of resistance genotypes among Beijing and non-Beijing strains was compared using a reverse blot hybridization assay to detect mutations within genes associated with rifampicin (rpoB) and isoniazid (katG, inhA, and ahpC) resistance. Of the 743 Mycobacterium tuberculosis isolates, 569 (77%) belonged to Beijing family. The proportion of Beijing strains was significantly higher among MDR-TB isolates than among drug-susceptible strains (82% vs. 72%, p<0.01). Genotype analysis of the rpoB gene revealed significantly lower rates of the Ser531Leu mutation rate among Beijing vs. non-Beijing MDR-TB strains (41% vs. 66%, p<0.005). While the mutation for Ser315Thr in the katG gene was more common among Beijing vs. non-Beijing family strains (65% vs. 50%, p<0.01), the mutation rate of promoter region of the inhA gene was lower among Beijing strains compared with non-Beijing strains (14% vs. 25%, p<0.05). Reverse hybridization successfully detected over 80% of isoniazid-resistant strains and over 92% of rifampicin-resistant strains among Korean isolates. Significant differences in mutation rates in the rpoB, katG, and inhA genes between Beijing strains and non-Beijing strains could explain discrepancies in mutation rates of genotypes in different countries. Reverse hybridization was useful for rapid detection of isoniazid and rifampicin resistant strains.
使用反向杂交分析法比较北京株和非北京株中耐药基因型的频率,以检测与利福平(rpoB)和异烟肼(katG、inhA和ahpC)耐药相关基因内的突变。在743株结核分枝杆菌分离株中,569株(77%)属于北京家族。耐多药结核分枝杆菌分离株中北京株的比例显著高于药物敏感株(82%对72%,p<0.01)。rpoB基因的基因型分析显示,北京耐多药结核分枝杆菌株中Ser531Leu突变率显著低于非北京株(41%对66%,p<0.005)。虽然katG基因中Ser315Thr突变在北京家族株中比非北京家族株中更常见(65%对50%,p<0.01),但北京株中inhA基因启动子区域的突变率低于非北京株(14%对25%,p<0.05)。反向杂交成功检测出韩国分离株中80%以上的异烟肼耐药株和92%以上的利福平耐药株。北京株和非北京株在rpoB、katG和inhA基因的突变率上存在显著差异,这可以解释不同国家基因型突变率的差异。反向杂交有助于快速检测异烟肼和利福平耐药株。