反向斑点杂交技术检测结核分枝杆菌3种耐药相关基因

J4 ›› 2011, Vol. 37 ›› Issue (4): 759-762.

反向斑点杂交技术检测结核分枝杆菌3种耐药相关基因

罗丹,向延根,潘建华,彭雪峰,邓为之,石国民

湖南省长沙市中心医院检验科|湖南长沙 410004

收稿日期:2010-12-23 出版日期:2011-07-28 发布日期:2011-07-28
通讯作者: 向延根(Tel:0731-85667943,E-mail:xiangyangen@126.com) E-mail:xiangyangen@126.com
作者简介:罗丹（1979-）|男|湖南省长沙市人|主管检验师|医学硕士|主要从事临床微生物和免疫方面的研究。
基金资助:
国家“十一五”科技重大专项资助课题（2009ZX10005-018）；湖南省卫生厅科技基金资助课题（B2008074）

Detection of |three-drug resistance related genes of mycobacterium tuberculosis by |membrance reverse dot blot hybridization

LUO Dan,XIANG Yan-gen,PAN Jian-hua,PENG Xue-feng,DENG Wei-zhi,SHI Guo-min

Department of Clinical Laboratory,Central Hospital of Changsha City,Hunan Province| Changsha 410004,China

Received:2010-12-23 Online:2011-07-28 Published:2011-07-28

摘要/Abstract

摘要：

目的：观察膜反向斑点杂交技术快速检测结核分枝杆菌（MTB）对异烟肼（INH）、利福平(RFP)、链霉素(SM)耐药性的效果。方法：用ropB、katG、inhA、rpsL基因的寡核苷酸探针与结核分枝杆菌PCR产物进行反向斑点杂交，并将结果与绝对浓度法药敏试验结果进行比较。结果：INH、RFP和SM的耐药基因检测灵敏度分别为84.88%、87.10%和79.17%，符合率为97.33%、94.74%和88.37%。结论：膜反向斑点杂交技术是检测部分MTB耐INH、RFP和SM基因型快速、简便的方法。

关键词: 结核分枝杆菌；药物耐受性；反向斑点杂交;聚合酶链式反应；基因

Abstract:

Abstract:Objective To observe the rapid detection effccts of resistance to isoniazide(INH),rifampcin (RFP)and streptomycin(SM) in mycobacterium tuberculosis (MTB) by membrance reverse dot blot hybridizd technique. Methods The probes of ropB,katG,inhA and rpsL resistant genes were mixed with PCR products of MTB .The mixture were detected by membrance reverse dot blot hybridization technigue.Then the results were compared with conventional drug susceptibility tests. Results The sensitivities of INH,RFP and SM were 84.88%,87.10% and 79.17%，and the coincident rates were 97.33%,94.74% and 88.37%. Conclusion The membrane-reverse dot blot hybridization can be used to detect resistance of partial MTB clinical strains rapidly and simply.

Key words: mycobacterium tuberculosis;drug resistance;reverse dot blot hybridization;polymerase chain reaction;gene

中图分类号:

罗丹,向延根,潘建华,彭雪峰,邓为之,石国民. 反向斑点杂交技术检测结核分枝杆菌3种耐药相关基因[J]. J4, 2011, 37(4): 759-762.

LUO Dan,XIANG Yan-gen,PAN Jian-hua,PENG Xue-feng,DENG Wei-zhi,SHI Guo-min. Detection of |three-drug resistance related genes of mycobacterium tuberculosis by |membrance reverse dot blot hybridization[J]. J4, 2011, 37(4): 759-762.