鳗鲡创伤弧菌的分子鉴定

目的对分离自发病欧洲鳗鲡的创伤弧菌疑似菌株进行准确的鉴定。方法首先尝试利用一对16SrRNA基因特异性通用引物PCR扩增一株鳗源创伤弧菌疑似株的基因组DNA，得到一个约500bp的DNA片段，将该DNA片段亚克隆至pMDl8-T载体，鉴定克隆化成功之后，送专业公司进行测序，得到一个502bp的DNA产物，NCBI上同源性比较表明，该片段与GeneBank上注册的创伤弧菌的16SrDNA序列同源性最高（100％），同时排除了哈维氏弧菌的可能。设计一对创伤弧菌溶血素特异性引物，实现了对鳗鲡创伤弧菌的分子鉴定。结果建立一种简洁的鳗鲡创伤弧菌的分子鉴定方法。结论国内首次自发病欧鳗分离到创伤弧菌，并给出分子鉴定证据。

Molecular identification of Vibrio vulnificus isolated from the diseased European eels in mainland China

To accurately identify the bacterial strains of Vibrio vulnificus isolated from the diseased European eels(Anguilla anguilla) in mainland of China,the genomic DNA from a suspected strain of V.vulnificus isolated from A.anguilla was amplified by PCR using a pair of universal primers specific to 16S rRNA gene.A DNA fragment about 500 bp in length was obtained,and this fragment was cloned into vector pMD18-T and sequenced after cloning.The DNA product of 502 bp in length was thus demonstrated later on.Compared with the NCMI homology,it was evident that the 16S rDNA sequence homology was the highest among those of V.vulnificus found in GenBank nucleotide sequence database with a sequence homology similarity of 100%,but low identity to V.harveyi was demonstrated,thereby excluding the possibility of V.harveyi.Also,a pair of primer specific to hemolysin/cytolysin gene was designed and was used for the identification of other vibrios.It was found from the results in PCR amplification that only V.vulnificus showed positive result,while a number of the pathogenic species of fish vibrios,such as V.harveyi,V.algonaticus,V.parahaemalyticus,V.anguillrum and Aeromonas hydrophila showed negative results.It is concluded that a simple molecular method for the accurate identification of V.vulnificus isolated from European eels was established,by which the first reported strain of V.vulnificus was isolated from diseased European eels in mainland of China..