【作者】 张建平； 沈兴华； 王霞芳； 唐佩军； 虞忻； 曹丽娟； 王雪峰； 吴妹英；

【Author】 ZHANG Jian-Ping~(1,2),SHEN Xing-Hua~(1,2),WANG Xia-Fang~2,TANG Pei-Jun~(1,2),YU xin~(1,2),CAO Li-Juan~2,WANG Xue-Feng~2,WU Mei-Ying~1. 1.The Affiliated Infectious Hospital of SuZhou University,Suzhou 215007,China;2.School of Biology and Basic Medical Sciences,SuZhou University,Suzhou 215007,China

【机构】 苏州大学附属传染病医院； 苏州大学生物与基础医学院；

【摘要】 目的了解苏州市地区耐利福平(Rifampin,RFP)结核分枝杆菌rpoB基因突变特点,以期为本地区RFP耐药结核分枝杆菌(Mycobacteriumtuberculosis,MTB)的临床快速诊断提供有价值的分子标记靶点。方法通过裂解MTB抽提基因组DNA,采用PCR扩增和DNA测序方法,分析苏州市结核病患者痰培养阳性并经药敏试验诊断为RFP耐药MTB的rpoB基因RFP耐药决定区(Rifampin resistance-determiningregion,RRDR)和CII区变异谱与变异率。将主要突变形式RRDR-531(TCG→TTG)与传统药敏实验进行诊断学试验,评价此类型突变作为RFP耐药菌株分子标记靶点的临床应用价值。结果 42例RFP耐药MTB的rpoB基因总突变率为90.5%(38/42),其中RRDR-516、RRDR-526、RRDR-531、RRDR-5 33和CII region-572突变率分别为7.1%(3/42)、19.0%(8/42)、64.3%(27/42)、2.4%(1/42)和2.4%(1/42),RRDR-533和CII region-572均以联合突变形式出现。在所检测到的突变中,以RRDR-531(TCG→TTG)为主要形式(61.9%)。以此类型的位点突变作为RFP耐药菌株诊断的分子标记,与传统药敏试验比较进行c~2检验,结果表明,与敏感菌株相比,RRDR-531(TCG→TTG)突变在RFP耐药菌株中呈显著差异(c~2=25.3,P<0.01),灵敏度为61.9%,特异度为100%,阳性预测值为100%,阴性预测值为60.98%,95%可信区间的比值比为81.91,似然比2.26。结论 rpoB基因RRDR-531(TCG→TTG)突变是苏州市地区MTB产生RFP耐药性的主要分子机制,其作为RFP耐药MTB诊断的分子标记靶点具有较好的临床应用价值。更多还原

【Abstract】 Objective To understanding the characteristics of rpoB gene mutation of rifampin(RFP) resistant Mycobacterium tuberculosis(MTB) isolated from tuberculosis patients in Suzhou China and the significance of potential typical mutation as a molecular marker to diagnose RFP-resistant MTB strains.Methods After dissolving MTB and amplifying rpoB gene fragment by PCR,direct sequencing was used to analyze the mutation spectrum occurred in rifampin resistance-determining region(RRDR) and C region of rpoB gene from 42 RFP-resistant and sensitive MTB strains.Diagnostic testing was used to evaluate the significance of typical mutation occurred in rpoB gene as a molecular marker of RFP-resistant MTB strains compared with the traditional sensitivity test.Results 38 of 42 RFP-resistant MTB strains(90.5%) was identified to contain base mutations occurred in RRDR an C region of rpoB gene.Among these,the mutation RRDR-531 TCG→TTG is the dominating form and occupied 61.9%.When the mutation RRDR-531 TCG→TTG was used to diagnose FRP-resistant MTB strains compared with the traditional sensitivity test as a marker,the mutation rate presented statistic significance (c~2=25.3 P<0.01) and the sensitivity,specificity,positive predictive value,negative predictive value,likelihood ratio and odds ratio with a 95%confidence interval was respectively revealed to be 61.9%,100%,100%,60.98%,2.26 and 81.91.Conclusion The mutation RRDR-531 TCG→TTG was revealed to be the primary mechanism of MTB resistance to RFP and presented the significance in diagnosing RFP-resistant MTB strains as a molecular marker used in Suzhou China.更多还原