16S rRNA在肉毒梭菌分型鉴定中的应用  

16S rRNA in identification and typing of Clostridium botulinum

作　　者：卢卫嘉[1] 毛晓燕[1] 熊颖[1] 张超[1] 王建锋[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《第二军医大学学报》2011年第11期1186-1188,共3页Academic Journal of Second Military Medical University

摘　　要：目的建立一种快速、可靠的对肉毒梭菌进行分型鉴定的手段。方法以从LCL063、830110、LC175、LCL155、66418、N153、61082、ALASKA、IWANAI共9株梭菌中提取的基因组DNA为模板,利用16SrRNA特异性引物分别进行PCR扩增并进行T-A克隆转化、测序。通过Clustal和Mega软件分析16SrDNA序列,以NJ法和MP法构建进化树,分析其种属特异性。结果 16SrRNA分型结果可判断出LCL063、830110、LCL175为产E型毒素的酪酸梭菌。IWANAI与ALASKA株为E型肉毒梭菌。与传统分型鉴定得到的结果一致。结论 16SrRNA在肉毒梭菌分型鉴定中具有快速、准确的优势,随着核糖体库的不断完善,有望成为细菌分型鉴定的标准依据。Objective To establish a rapid,reliable method for identifying and typing of Clostridium botulinum using 16S rDNA sequencing and phylogenetic tree construction.Methods Clostridium genome templates were extracted from 9 strains,including LCL063,830110,LC175,LCL155,66418,N153,61082,ALASKA,and IWANAI;16S rRNA genes were amplifed by PCR with the 16S rRNA specific primers,and then the PCR products were cloned to pGEM-T Easy vector and sequenced.Finally,the sequence of 16S rDNA was analyzed by Clustal and Mega program;the phylogenetic trees were constructed by Neighor joining and Maximum parsimony.Results It was found that LCL063,830110,and LCL175 were BONT/E producing Clostridium butyricums;IWANAI and ALASKA were Clostridium botulinum type E.The results of the present method were consistent with those of the conventional method.Conclusion 16S rRNA sequencing combined with phylogenetic tree analysis is a rapid and accurate method in Clostridium botulinum identification,and the method may serve as a criterion for bacterial typing with the completion of ribosomal RNA data bank.

关 键 词：16SrRNA 肉毒梭菌 进化树 细菌分型技术 

分 类 号：R372[医药卫生—病原生物学]