可疑霍乱弧菌的实验鉴定

目的:对4株可疑霍乱弧菌进行种属鉴定。方法:对可疑菌落在传统血清学和系统生化鉴定的基础上进行细菌的分子鉴定:应用16S rDNA的序列分析进行细菌种属鉴定和实时荧光PCR检测O1、O139群霍乱弧菌。结果:16 s rDNA基因序列分析显示041和067为麦氏弧菌,059和074为霍乱弧菌;荧光定量PCR检测结果说明4株菌都不是O1和O139霍乱弧菌。结论:结合血清凝集和系统生化实验,可以确定041和067为麦氏弧菌,059和074为非O1/非O139霍乱弧菌。对于可疑和难鉴别的霍乱弧菌,不能单纯依靠血清学和表型鉴定方法,需要分子诊断方法,尤其是16SrDNA的序列分析,可以从分子水平为弧菌科细菌的种属鉴定提供更为直接的证据。

Experimental identification of suspected Vibrio cholerae isolates

Objective:To identify four suspected Vibrio cholerae isolates.Methods:Detect four suspected V.cholerae strains using molecular technology including sequence analysis of 16S rDNA on the bacterial species level and TaqMan-based Real-time PCR for V.cholerae O1 and O139,on the basis of classic serological and biochemical assays.Results:16SrDNA sequencing revealed that strains 041 and 067 belong to Vibrio metschnikovii,whereas the other two isolates 059 and 074 belong to V.cholerae.None of four isolates were proved to be V.cholerae O1 or O139 by real-time PCR.Conclusion:Two of four isolates were identified as V.metschnikovii and the other as V.choleare non-O1/non-O139 by the combination of serological and biochemical experiments.To detect suspected and non-classified V.cholerae strains,molecular assay are required to exclude obscure factors from phenotypic identification,especially direct sequencing of 16SrDNA which can support for accurate identification of Vibrio species.