建立PC-PCR法快速、半定量检测养殖环境和水产动物中的致病性副溶血弧菌

›› 2008, Vol. 10 ›› Issue (S1): 67-72.

建立PC-PCR法快速、半定量检测养殖环境和水产动物中的致病性副溶血弧菌

马妍李健王群何玉英王斌

大连水产学院生命科学与技术学院中国水产科学研究院黄海水产研究所农业部海洋渔业资源可持续利用重点开放实验室大连水产学院生命科学与技术学院辽宁大连中国水产科学研究院黄海水产研究所农业部海洋渔业资源可持续利用重点开放实验室山东青岛辽宁大连

收稿日期:2008-02-27 修回日期:2008-03-23 出版日期:2008-05-31 发布日期:2009-07-29
通讯作者: 李健,研究员,主要从事海水健康养殖和水产药理研究。
作者简介:马妍, 硕士研究生,主要从事海洋致病微生物评估研究。
基金资助:
国家863计划项目(2006AA10A406);; 国家科技支撑计划项目(2006BAD01A13);; 公益性农业行业科研专项(nyhyzx07-042)资助

Rapid and Semi-quantitative Detection of Pathogenic Vibrio paraharmolyticus from Environment and Aquatic Animals by PC-PCR

1.College of Life Sciences and Biotechnology|Dalian Fisheries University|Liaoning Dalian 116023|2.Key Opening Laboratory of Agricutural Ministry for Marine fishery Resources Sustainable Ultilization|Yellow Sea Fisheries Research Institute|Chinese Academy of Fishery Sciences|Shandong Qingdao 266071|China

Received:2008-02-27 Revised:2008-03-23 Online:2008-05-31 Published:2009-07-29

摘要/Abstract

摘要：

以tl为靶基因,将菌落PCR技术和平板菌落计数法相结合,建立致病性副溶血弧菌快速、半定量检测方法。PCR扩增的目的片段为673 bp,人工污染样品检测最低限固体样品为3.1×102cfu/g,液体样品为1.3×102cfu/mL,菌落PCR每反应体系的检测低限为90 cfu。对人工污染样品的检测结果显示,PC-PCR方法检测结果与常规检测结果一致,可用于养殖环境及水产品中细菌的动态监测。

关键词: 副溶血弧菌快速检测半定量菌落PCR tl基因

Abstract:

A rapid and semi-quantitative method was developed for detection of pathogenic Vibrio paraharmolyticus,using tl gene as a targeted gene,combined the method of plate count for bacterial colonies and colony PCR.The expected PCR product was 673 bp.Detection limit for artificial contaminants was 3.1×102 cfu/g of solid sample and 1.3 102 cfu/ml of liquid sample;Detection limit of colony PCR reaction system was 90 cfu.The contaminated samples were detected by both PC-PCR and routine methods with a completely coin...

Key words: Vibrio parahaemolyticus, rapid detection, semi-quantitative, colony PCR, tl gene;

中图分类号:

S941

马妍,李健,王群,何玉英,王斌. 建立PC-PCR法快速、半定量检测养殖环境和水产动物中的致病性副溶血弧菌[J]. , 2008, 10(S1): 67-72.