| 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16S rRNA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16S rDNA序列,以NJ法和MP法构建进化树,分析其种属特异性。 结果 16S rRNA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 16S rRNA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。 |
| 关键词: 16S rRNA 肉毒梭菌 进化树 细菌分型技术 |
| DOI：10.3724/SP.J.1008.2011.01186 |
| 投稿时间：2011-08-15修订日期：2011-10-09 |
| 基金项目: |
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| 16S rRNA in identification and typing of Clostridium botulinum |
| LU Wei-jia,MAO Xiao-yan*,XIONG Ying,ZHANG Chao,WANG Jian-feng |
(Lanzhou Institute of Biological Products, Lanzhou 730046, Gansu, China
*Corresponding author.) |
| Abstract: |
| 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. |
| Key words: 16S rRNA Clostridium butyricum phylogenetic tree bacterial typing techniques |
