PCR快速检测水产品中副溶血性弧菌的方法研究.docVIP

2008, Vol. 29, No. 11 食品科学※分析检测 490PCR 快速检测水产品中副溶血性弧菌的 方法研究 刘 琦1, 2，周德庆1，柳淑芳1，苏来金1 (1.中国水产科学研究院黄海水产研究所，山东 青岛 266071；2. 上海海洋大学食品学院，上海 200090摘 要：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t l 基因设计合成引物，采用酚-氯仿法提取D N A ，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。关键词：副溶血性弧菌；水产品；检测 Study on PCR Method for Rapid Detection of Vibrio parahaemolyticus in Aquatic Products LIU Qi1,2，ZHOU De-qing1，LIU Shu-fang1，SU Lai-jin1 (1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China； 2. College of Food Science, Shanghai Ocean University, Shanghai 200090, China Abstract ：In order to improve the detection efficiency of Vibrio parahaemolyticus in aquatic products to meet the need of asystem of rapid reaction for food sanitationlas PCR method was established. A primer in connection with t1 gene which is specificfor Vibro parahaemolyticus was designed and synthesized, phenol-chloroform method was used to extract DNA, and controlsfor method specificity were made with Escherichia coli, Salmonella , Shigella , Vibrio alginolyticus. 40 aquatic products sampledat 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 liquid103 CFU/ml. The samples with bacteria amount of 1～10 CFU/g can be detected after enrichment of 6 h with a detection cycleof 10 h. Results of this method are more accurate and more reliable compared with national standard. So this method is suitablefor rapid detection of Vibrio parahaemolyticus in aquatic products.Key words：Vibro parahaemolyticus；aquatic products；detec