检测水产品食物中副溶血弧菌基因分型方法的建立

Establishment of A Molecular Typing Method of Vibrio parahaemolyticus for Testing of Food Poisoning

目的建立副溶血弧菌的基因分型技术,应用于广州市水产品食物中毒居前列的致病菌——副溶血弧菌的检测,寻找一种简便快速准确的检验方法。方法分析副溶血性弧菌分离株的16S-23S rDNA IGS的相关信息(电泳图谱、序列分析等),找出合适的16S-23S rDNA IGS作为基因分型依据,建立副溶血性弧菌的基因分型技术。结果通过克隆和测序,分析了副溶血性弧菌的16S-23S rDNA IGS片段,初步确定了以16S-23S rDNA IGS为分子标记的副溶血性弧菌基因分型技术。结论本检测方法不但快速、灵敏,还对深入了解副溶血性弧菌的发病机制,研究其分子流行病学特征具有重要的意义。

Objective To develop a genotyping method of Vibrio parahaemolyticus, which is one of the most frequent pathogenic bacterium in food poisoning in Guangzhou, for quick and accurate testing of food poisoning agent. Methods The electrophoresis pattern and nucleotide sequence of 16S-23S rDNA IGS were analyzed. The suitable target DNA segment for genotyping of Vibrio parahaemolyticus was selected. The molecular typing procedure was established based on the target sequence. Results A 16S-23S rDNA IGS fragment was successfully cloned and sequenced. Molecular typing method of Vibrio parahaemolyticus was successfully developed based on the molecular marker of 16S- 23S rDNA IGS. Conclusion The genotyping of Vibrio parahaemolyticus based on the 16S-23S rDNA IGS was not only a quick but also a sensitive method, which offered a new method for testing food poisoning based on genotyping. And this may play an important role on understanding the pathogenesis of Vibrio parahaemolyticus and its molecular epidemiology.