冰鲜金鲳鱼样品中一株副溶血性弧菌的分离与鉴定
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。
Objective To analyze and identify a bacterium which isolated from ice fresh Trachinotus ovatus sample. Methods A bacterium named sznjV083 was isolated from ice fresh Trachinotus 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 was Vibrio vulnificus. Real-time PCR showed that it was Vibrio parahaemolyticus. The results about sequence analysis of 16S rDNA and phy-logenic tree showed that sznjV083 was very closed to Vibrio Parahaemolyticus BB22OP (Accession CP003973.1). The virulence genes tdh and trh were detected as negative. Conclusion Comprehensive analysis of the results of real-time PCR and 16S rDNA sequence analysis, bacterium sznjV083 which isolated from ice fresh Trachinotus ovatus sample was belonged to Vibrio 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.
标题：冰鲜金鲳鱼样品中一株副溶血性弧菌的分离与鉴定
英文标题：Isolation and identification of Vibrio parahaemolyticus from iced fresh Tra-chinotus ovatus
作者：
柯艳坤 深圳市农产品质量安全检验检测中心
阚式绂 深圳市农产品质量安全检验检测中心
钟雷响 深圳市农产品质量安全检验检测中心
汪清 深圳市农产品质量安全检验检测中心
罗国强 深圳市农产品质量安全检验检测中心
但琨 深圳市农产品质量安全检验检测中心
中文关键词:副溶血性弧菌,生理生化特征,PCR,16S rDNA序列,系统发育树,
英文关键词:Vibrio parahaemolyticus,physiological and biochemical characteristics,PCR,16S rDNA,phylogenic tree,
发表日期：2015-05-19
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