PCR快速检测水产品中副溶血性弧菌的方法研究

Study on PCR Method for Rapid Detection of Vibrio parahaemolyticus in Aquatic Products

为了满足食品卫生快速反应体系的需要,提高水产品中副溶血性弧菌(Vibrio parahaemolyticus,VP)的检测效率,本实验针对副溶血性弧菌特异性tl基因设计合成引物,采用酚-氯仿法提取DNA,利用大肠杆菌(Escherichia coli)、沙门氏菌(Salmonella)、志贺氏菌(Shigella)、溶藻弧菌(Vibrio alginolyticus)做方法特异性对照,建立了水产品中副溶血弧菌PCR快速检测方法。利用此方法对市场随机抽取的40份水产品进行检测,并用国家标准方法(GB/T4789.7—2003)对该方法的检验结果进行评价。结果表明:该方法操作简便,特异性强,菌液灵敏度为103CFU/ml,含菌量1～10CFU/g的样品经增菌6h后即可检出。与国标方法相比更加准确可靠,检测周期10h即可完成。说明此方法适合水产品中副溶血性弧菌的快速检测。

In order to improve the detection efficiency of Vibrio parahaemolyticus in aquatic products to meet the need of a system of rapid reaction for food sanitationlas PCR method was established. A primer in connection with tl gene which is specific for Vibro parahaemolyticus was designed and synthesized, phenol-chloroform method was used to extract DNA, and controls for method specificity were made with Escherichia coli, Salmonella, Shigella, Vibrio alginolyticus. 40 aquatic products sampled at random from market were tested by using this method, and the results were evaluated with national standard （GB/T 4789.7 --2003）. Results showed that this method is easy for operation, with high level of specificity and the sensitivity of bacterial liquid 103 CFU/ml. The samples with bacteria amount of 1 ～ 10 CFU/g can be detected after enrichment of 6 h with a detection cycle of 10 h. Results of this method are more accurate and more reliable compared with national standard. So this method is suitable for rapid detection of Vibrio parahaemolyticus in aquatic products.