食品中五种致病性弧菌MPCRDHPLC检测方法的建立.pdfVIP

500 2008, Vol. 29, No. 10 食品科学 ※分析检测 1 2 1 1 , 3 1 1 邵碧英 ，王传得 ，郑 晶 ，黄晓蓉 * ，曹际娟 ，陈 彬 ，吴 谦 (1.福建出入境检验检疫局，福建 福州 350001 ；2.福建农林大学动物科学院，福建 福州 350002 ； 3. 辽宁出入境检验检疫局，辽宁大连 116001) ：采用试剂盒法提取5 种致病性弧菌(霍乱弧菌、副溶血性弧菌、创伤弧菌、溶藻弧菌和拟态弧菌) 的基因 组DNA 。分别合成5 对特异性引物，对PCR 反应体系进行优化后，建立了5 种致病性弧菌间的MPCR-DHPLC 检 测方法。二重PCR-DHPLC 检测灵敏度可达到100CFU/ml。建立的MPCR-DHPLC 方法在食品中弧菌的检测上具有 很好的应用价值。 ：弧菌；基因组DN A ；多重PC R ；变性高效液相色谱 Development of MPCR-DHPLC Detection Method for Five Pathogenic Vibrios in Foods 1 2 1 1,* 3 1 1 SHAO Bi-ying ，WANG Chuan-de ，ZHENG Jing ，HUANG Xiao-rong ，Cao Ji-juan ，CHEN Bin ，WU Qian (1.Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou 350001, China ； 2.College of Animal Science ，Fujian Agriculture and Forestry University, Fuzhou 350002, China ； 3.Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China) Abstract ：The genome DNAs of five pathogenic Vibrios were extracted by kit method, including Vibrio cholerae, V. parahaemolyticus , V. vulnificus, V. alginolyticus and V. mimicus, and their five pairs of special primers were composed, respectively. The MPCR-DHPLC detection methods among the five vibrios were developed after the PCR reaction systems were optimized. The detection sensitivity of duplex PCR-DHPLC is 100 CFU/ml. The developed MPCR-DHPLC method has good application merit on detecting the Vibrios in foods. Key words ：Vibrio；genome DNA ；multiplex PCR (MPCR)；denaturalization high performance liquid chromatography (DHPLC) 中图分类号：Q93-332 文献标识码：A