Email Alert | RSS    帮助

中国防痨杂志 ›› 2011, Vol. 33 ›› Issue (10): 680-685.

• 论著 • 上一篇    下一篇

应用基因芯片方法检测结核分枝杆菌利福平和异烟肼的耐药性

张俊仙,吴雪琼,阳幼荣,梁艳   

  1. 北京,解放军第三〇九医院全军结核病研究所  军队结核病防治重点实验室
  • 收稿日期:2011-06-12 出版日期:2011-10-10 发布日期:2012-03-07
  • 通信作者: 吴雪琼 E-mail:wu-xueqiong@263.net
  • 基金资助:

    北京市科技重大专项重点项目(Z09050700940904);“十一五”国家重大科技专项(2008ZX10003-001)

ZHANG Jun-xian, WU Xue-qiong, YANG You-rong, LIANG Yan   

  1. Army Tuberculosis Prevention and Control Key Laboratory, Institute for Tuberculosis Research, The 309th Hospital of Chinese People’Liberation Army, Beijing 100091, China
  • Received:2011-06-12 Online:2011-10-10 Published:2012-03-07
  • Contact: WU Xue-qiong E-mail:wu-xueqiong@263.net

摘要: 目的 应用基因芯片方法检测结核分枝杆菌(Mycobacterium tuberculosis,Mtb)对利福平和异烟肼的耐受性,评价其临床应用价值。方法 应用聚合酶链反应(polymerase chain reaction,PCR)扩增-基因芯片杂交的方法检测经常规药敏实验证实的30株Mtb利福平和异烟肼敏感株和50株耐利福平和异烟肼分离株的rpoB基因及katGinhA基因突变,同时以PCR-直接测序法为对照。结果 应用PCR-基因芯片与基因测序方法检测30株Mtb利福平敏感株rpoB基因和异烟肼敏感株katG基因和inhA基因均为野生型。50株Mtb利福平耐药株中,PCR-基因芯片与基因测序分析3株rpoB基因均为野生型,41株均为突变型;6株PCR-基因芯片与基因测序结果不一致。50株Mtb异烟肼耐药株中,PCR-基因芯片与基因测序分析16株katG基因和30株inhA基因均为野生型,31株katG基因均为315位密码子突变,7株inhA基因均为15位突变型,其中2株为katGinhA双重突变;3株katG和13株inhA PCR-基因芯片与基因测序结果不一致。结论 应用PCR-基因芯片方法可快速、有效地检出大多数Mtb耐多药分离株,指导临床用药。

关键词: 分枝杆菌, 结核, 利福平, 异烟肼, 抗药性, 细菌, 寡核苷酸序列分析

Abstract: Objective To detect Mycobacterium tuberculosis(Mtb)resistance to rifampicin (RIF) and isoniazid (INH) by gene chip,and to evaluate their clinical values. Methods Thirty RIF- and INH-susceptible and 50 RIF- and INH-resistant clinical isolates in Mtb were analyzed the mutations of rpoB、katG and inhA genes by (PCR) gene chip and DNA sequencing.  Results The rpoB、katG and inhA genes from 30 RIF and INHsusceptible isolates of Mtb were all wild types by PCRgene chip and DNA sequencing. Of 50 RIFresistant clinical isolates, 3 had wild types of ropB genes, and 41 had ropB mutations by PCRgene chip and DNA sequencing. Six had inconsistent results between PCRgene chip and DNA sequencing. Of 50 INHresistant clinical isolates, 16 had wild types of katG genes, and 31 had katG mutations on codon 315; 30 had wild types of inhA genes, and 7 had inhA mutations at position15. KatG results of 3 isolates and inhA results of 13 isolates were inconsistent between PCRgene chip and DNA sequencing. Conclusion PCR-gene chip might be a rapid and effective method for the detection of Mtb multi-drug-resistant isolates.

Key words: Mycobacterium tuberculosis, Rifampicin, Isoniazid, Drug resistance,bacterial, Oligonucletide array sequence analysis