摘要
目的了解结核分支杆菌链霉素(Sm)耐药基因突变情况,建立快速检测耐药突变株的方法。方法通过聚合酶链反应-单链构象多态性(PCR-SSCP)分析了22株结核分支杆菌临床分离株的rpsL基因,并应用PCR-限制性片段长度多态性(PCR-RFLP)进一步分析其突变的部位和性质。结果以H37Rv标准株为对照,5株敏感株的rpsL基因未见SSCP泳动异常、有MboⅡ酶切位点;13株耐Sm株中,11株(86%)有rpsL基因泳动异常、无MboⅡ酶切位点;4株耐其它抗结核药物株中,也有1株SSCP泳动异常、并无MboⅡ酶切位点。结论大多数结核分支杆菌Sm耐药分离株有rpsL突变,而突变位点大多位于第43位密码子。应用PCR-SSCP和PCR-RFLP可能成为快速检测结核分支杆菌耐药突变株的新方法。
ObjectiveToobservethemutationsofrpsLgeneinM.tuberculosisstreptomycin-resis-tantisolates,andtodevelopanewmethodfordetectingdrugresistance.MethodDetectingtherpsLasacontrol.In22M.tuberculosisclinicalisolates,therpsLPCR-RFLP.ResultsStrainH37Rvwasusedasacontrol.In22M.tuberculosisclinicalisolates,therpsLPCRfragmentsfrom5drug-suscep-tibleisolateshadnodiferencesintheSSCPprofileswithstrainH37Rv,andwererestrictedbyMboⅡ.11ofthe13streptomycin-resistantisolatesshowedapparentdiferencesintheSSCPprofilesandwerenotdigestedwithMboⅡ.1ofthe4otherdrug-resistantisolatesalsohadapparentSSCPdiferencesandwasnotdigestedbyMboⅡ.ConclusionsTheresultssuggestedthattherpsLgenemutationwasfrequantlyobservedinM.tuberculosisstreptomycin-resistantisolates,andusualysituatedatcodon43,PCR-SS-CPandPCR-RFLPmightbecomeasimple,rapidandreliablediagnostictestfordrugre-sistance.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
1996年第6期342-345,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
关键词
结核分支杆菌
聚合酶链反应
药物耐受性
MycobacteriumtuberculosisPolymeraseChainReactionPolymorphism,single-strandedconformationalPolymorphism,restrictionfragmentlengthDrugtolerance