PCR-SSCP分支杆菌菌种初步鉴定方法的建立及其应用  

DIFFERENTIATION OF MYCOBACTERIAL SPECIES BY PCR-SSCP

作　　者：吴雪琼[1] 张俊仙[1] 刘佳文 黄海南 王晋洪[2] 

机构地区：[1]解放军309医院结核病研究室,北京100091 [2]山东省胸科医院检验科

出　　处：《中国现代医学杂志》2000年第7期25-26,共2页China Journal of Modern Medicine

摘　　要：建立一种简便、快速、灵敏和特异的分支杆菌菌种鉴定方法。方法 :通过聚合酶链反应 (PolymeraseChainReaction ,PCR) -单链构象多态性 (Single -strndedConformationPolymorphism ,SSCP)方法分析分支杆菌 16SrRNA ,根据其电泳图谱与标准株的相似性进行菌种初步鉴定。结果 :2 9种分支杆菌标准株中 ,除结核分支杆菌、牛结核分支杆菌和卡介苗之间及堪萨斯分支杆菌和胃分支杆菌之间SSCP电泳图谱相似外 ,其余各种分支杆之间电泳图谱均有显著差异。 70株BACTECTB4 60初步鉴定为结核分支杆菌复合群的临床分离株中 ,55株显示与结核分支杆菌SSCP电泳图谱相似 ,15例 (2 1.4 % )显示出与结核分支杆菌完全不同的电泳图谱 ;10株经传统方法鉴定为结核分支杆菌的分离株中 ,仅 1株显示出与结核分支杆菌不同的电泳图谱。结论 :PCR -SSCP有可能成为一种快速、有效的分支杆菌菌种初步鉴定方法。Objective: To develop a simple, rapid, sensitive, and specific method for differenciation of mycobacterial species. Methods: Mycobacterial 16S rRNSA sequences were analyzed by PCR-SSCP, then mycobacterial species were differentiated by their characteristic electrophoresis patterns. Results: Of 29 mycobacterial standard strains, M. tuberculosis, M. bovis and M. bovis BCG had similar SSCP profiles, M. kansassi and M. gastri also exhibited the same SSCP profiles, the other mycobacterial standard strains displayed different SSCP profiles. Of 70 clinical isolate identified as M. tuberculosis complex by BACTEC 460, 55 had the same SSCP profiles as M. tuberculosis, 15 exhibited different SSCP profiles from M. tuberculosis. Of 10 clinical isolate identified as M. tuberculosis by traditional method, only one exhibited different SSCP profiles from M. tuberculosis. Conclusions: PCR-SSCP technique might become a simple, rapid and reliable method for identification of mycobacterial species.

关 键 词：菌种鉴定 分支杆菌 PCR-SSCP 

分 类 号：R378.91[医药卫生—病原生物学]