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 and reliable means of typing and identification of Clostridium botulinum. Methods Genomic DNA extracted from 9 Clostridium strains of LCL063, 830110, LC175, LCL155, 64188, N153, 61082, ALASKA, and IWANAI was used as a template for PCR amplification and TA cloning using 16SrRNA specific primers, . The 16S rDNA sequences were analyzed by Clustal and Mega software. The phylogenetic tree was constructed by NJ method and MP method, and its species specificity was analyzed. Results 16S rRNA typing results can be judged LCL063, 830110, LCL175 to produce type C. toxin Clostridium butyricum. IWANAI and ALASKA strains are Clostridium botulinum type E. Consistent with the results of traditional typing. Conclusion 16SrRNA has a rapid and accurate advantage in the typing and identification of Clostridium botulinum. As the ribosomal library continues to improve, 16SrRNA is expected to become the standard basis for bacterial typing.