摘要
目的通过药物敏感性试验(简称“药敏试验”)获得结核分枝杆菌(MTB)临床分离株对贝达喹啉(Bdq)的最小抑菌浓度(minimum inhibitory concentration,MIC),并从基因水平分析MTB对Bdq的耐药机制。方法MTB菌株分离来自NDIP(New Drug Introduction Protection)项目合作医院的189例肺结核患者,其中,130例患者仅提供基线(服用Bdq第1剂前1周内)培养阳性菌株,59例除提供基线菌株外还提供开始含Bdq方案治疗后第2、4、8、12、16、20、24周时收集的培养阳性菌株。采用CMP1BDQ 96孔药敏微孔板检测患者基线MTB分离株对Bdq及另外11种抗结核药品耐药情况,并检测患者开始含Bdq方案治疗后不同时期的MTB临床分离株对Bdq的耐药情况。筛选出对Bdq耐药的MTB菌株,检测其Rv0678、atpE、pepQ、Rv1979c等4个基因的序列。结果86.2%(163/189)的基线MTB分离株对Bdq的MIC值分布在0.06μg/ml以下,MIC50和MIC90分别为0.03μg/ml和0.12μg/ml;基线MTB分离株对Bdq的原发耐药率为2.1%(4/189);患者在治疗过程中产生的Bdq获得性耐药率为1.6%(3/189)。4例对Bdq原发耐药患者MTB分离株均检测出Rv0678突变,未检出atpE、pepQ和Rv1979c突变;3例Bdq获得性耐药患者MTB分离株均检测出Rv0678突变,1例检测出Rv1979c突变,未检出pepQ和atpE突变;3例对Bdq敏感性降低患者MTB分离株中有2株检测出Rv0678突变,1株检测出Rv1979c突变,未检出atpE和pepQ突变。结论目前MTB临床分离株对Bdq的耐药率较低,耐药突变均发生在已知基因,未发现新的耐药相关基因。
Objective To explore the minimal inhibitory concentration(MIC)of clinical isolates of Mycobacterium tuberculosis(MTB)to bedaquiline(Bdq)by drug sensitivity test,and to analyze the mechanism of drug resistance of MTB to Bdq at the gene level.Methods MTB strains were isolated from 189 pulmonary tuberculosis patients from the cooperative Hospital of NDIP(New Drug Introduction Protection)project.Among them,130 patients only provided the baseline(within one week before taking the first dose of Bdq)positive strains,and the other 59 also provided the positive strains collected at 2,4,8,12,16,20 and 24 weeks after being treated with Bdq.CMP1BDQ 96 well drug sensitive microplate was used to detect the drug resistance of baseline MTB isolates to Bdq and other 11 kinds of anti-tuberculosis drugs,and the drug resistance of MTB clinical isolates to Bdq in different periods after being treated with Bdq regimen.MTB strains resistant to Bdq were screened and sequences of four genes,Rv0678,atpE,pepQ and Rv1979c,were detected.Results MIC values of 86.2%(163/189)of the baseline MTB isolates were below 0.06μg/ml,MIC50 and MIC90 were 0.03μg/ml and 0.12μg/ml,respectively;the primary resistance rate of baseline MTB isolates to Bdq was 2.1%(4/189);the acquired resistance rate of patients to Bdq during the treatment was 1.6%(3/189).Rv0678 mutation was detected in all of the 4 MTB isolates from patients with primary resistance to Bdq,but no atpE,pepQ,or Rv1979c mutations were detected;Rv0678 mutation was detected in MTB isolates from 3 patients with acquired resistance to Bdq,and Rv1979c was detected in 1 case,pepQ or atpE mutations were not detected;of the 3 patients with reduced sensitivity to Bdq,2 of the MTB isolates were detected with Rv0678 mutation,1 was detected with Rv1979c mutation,and atpE or pepQ mutations were not detected.Conclusion At present,the drug resistance rate of clinical isolates of MTB to Bdq is low,and drug resistance mutations all occur in known genes,no new drug resistance-related genes have been found.
作者
王璐
薛仲探
王玉峰
尚媛媛
任卫聪
姚丛
高飞
逄宇
WANG Lu;XUE Zhong-tan;WANG Yu-feng;SHANG Yuan-yuan;REN Wei-cong;YAO Cong;GAO Fei;PANG Yu(Graduate School of Inner Mongolia Medical University,Hohhot 010059,China;不详)
出处
《中国防痨杂志》
CAS
CSCD
2021年第5期482-486,共5页
Chinese Journal of Antituberculosis
基金
北京市医院管理中心“登峰”人才培养计划(DFL20191601)
北京市医院管理中心“扬帆”计划(ZYLX202122)。
关键词
分枝杆菌
结核
微生物敏感性试验
抗药性
基因
贝达喹啉
Mycobacterium tuberculosis
Microbial sensitivity tests
Drug resistance
Genes
bedaquiline
