冰鲜金鲳鱼样品中一株副溶血性弧菌的分离与鉴定

摘要

Objective To analyze and identify a bacterium which isolated from ice freshTrachinotus ovatus sample.MethodsA bacterium named sznjV083 was isolated from ice freshTrachinotus ovatus sample on TCBS and vibrio chromogenic medium, and then physiological and biochemical characteristics were characterized, and molecular biological characteristics were analyzed. At the same time, the standard strain ATCC33847 (Vibrio parahaemolyticus) and standard strain ATCC27562 (Vibrio vulnificus) were tested for quality control. Results Its colonies features on TCBS were blue-green and on vibrio chromogenic medium were dark-blue. VITEK 2 Gram-Negative identification card showed that it wasVibrio vulnificus. Real-time PCRshowed that it wasVibrio parahaemolyticus. The results about sequenceanalysis of 16S rDNA and phylogenic tree showed that sznjV083 was very closed toVibrio ParahaemolyticusBB22OP (Accession CP003973.1).The virulence genes tdhandtrh were detected as negative. ConclusionComprehensive analysis of the results of real-time PCR and 16S rDNA sequenceanalysis, bacterium sznjV083 which isolated from ice freshTrachinotus ovatus sample was belonged toVibrio parahaemolyticus. In the detection of Vibrio parahaemolyticus, in addition to the conventional physiological and biochemical methods, the experiment should also improve the specificity and sensitivity of detection results according to molecular biological method, such as the 16S rDNA gene sequence analysis or PCR method.%目的：对一株分离于冰鲜金鲳鱼样品中的菌株进行分析鉴定。方法利用TCBS培养基和弧菌选择性培养基从冰鲜金鲳鱼样品中分离到1株菌株sznjV083,并对该菌株的菌落形态、生理生化特征及其分子生物学特性进行分析。同时进行副溶血性弧菌(Vibrio parahaemolyticus)标准菌株ATCC33847,创伤弧菌(Vibrio vulnificus)标准菌株ATCC27562的质控检验。结果该菌株在TCBS平板上呈现蓝绿色菌落,而在弧菌显色平板上呈现深蓝色菌落,其全自动生化鉴定结果显示其为创伤弧菌。PCR试验和16S rDNA序列分析表明,该菌株为副溶血性弧菌并与参考菌株Vibrio parahaemolyticus BB22OP(登陆号CP003973.1)的同源性最高。且该菌株的毒力基因tdh、trh检测为阴性。结论分离于冰鲜金鲳鱼样品中的菌株sznjV083为副溶血性弧菌。在副溶血性弧菌的检测中,除了常规的生理生化等方法外,还应该结合16S rDNA基因序列分析或PCR方法提高检测结果的特异性和灵敏度,以保证检测结果的真实性和准确性。