| 水产动物致病性副溶血弧菌双重PCR 检测方法的研究 |
| |
| 引用本文： | 张晓君,梁利国,阎斌伦,秦 蕾,戚耀之.水产动物致病性副溶血弧菌双重PCR 检测方法的研究[J].海洋科学,2010,34(10):7-12. |
| |
| 作者姓名： | 张晓君 梁利国 阎斌伦 秦 蕾 戚耀之 |
| |
| 作者单位： | 淮海工学院,海洋学院,江苏省海洋生物技术重点建设实验室,江苏,连云港,222005 |
| |
| 基金项目： | 江苏省水产三项工程项目(PJ2010-58); 江苏省自然科学基金项目(BK2009163); 淮海工学院江苏省海洋生物技术重点建设实验室研究基金项目(2008HS016) |
| |
| 摘 要： | 副溶血弧菌(Vibrio parahaemolyticus)是多种水产养殖动物的主要病原菌,尤其是可引起凡纳滨对虾幼虾呈毁灭性死亡。本研究基于gyrB和toxR两种基因序列设计2对特异性引物,建立一种副溶血弧菌快速、准确的双重PCR检测方法,扩增目的片段大小分别为285 bp和368 bp。结果表明在同一PCR反应体系中副溶血弧菌可同时扩增大小分别为285 bp和368 bp的2种基因片段,两种引物对4种其他水产动物病原菌无交叉反应;敏感性检测结果显示,该双重PCR最低能检测8.867 2×103 CFU/mL菌体浓度的副溶血弧菌,对副溶血弧菌模板DNA的检出极限为0.029 3 mg/L;对发病中国对虾糠虾幼体、水产品及虾池养殖用水进行双重PCR检测,呈阳性反应的样品可分离出优势生长的副溶血弧菌。该实验所建立的基于gyrB和toxR两种基因的双重PCR检测方法可用于副溶血弧菌引起的水产动物疾病的快速诊断及分子流行病学的调查研究。
|
| 关 键 词： | 副溶血弧菌(Vibrio parahaemolyticus) gyrB 基因 toxR 基因 双重PCR |
| 收稿时间： | 2009/12/4 0:00:00 |
| 修稿时间： | 2010/1/11 0:00:00 |
Detection of pathogenic Vibrio parahaemolyticus isolated from aquatic animals by dulplex PCR |
| |
| ZHANG Xiao-jun,LIANG Li-guo,YAN Bin-lun,QIN Lei and QI Yao-zhi.Detection of pathogenic Vibrio parahaemolyticus isolated from aquatic animals by dulplex PCR[J].Marine Sciences,2010,34(10):7-12. |
| |
| Authors: | ZHANG Xiao-jun LIANG Li-guo YAN Bin-lun QIN Lei and QI Yao-zhi |
| |
| Institution: | ZHANG Xiao-jun,LIANG Li-guo,YAN Bin-lun,QIN Lei,QI Yao-zhi(College of Ocean,Key Laboratory of Oceanic Biotechnology of Jiangsu,Huaihai Institute of Technology,Lianyungang 222005,China) |
| |
| Abstract: | V. parahaemolyticus is a significant pathogen for the aquaculture industry, particularly devastating impact on cultivated Litopenaeus vannamei. In this study, two pair of specific primers was designed that allowed amplification of 285 bp and 368bp gene fragments based on gyrB and toxR genes. A simple dulplex polymerase chain reaction (PCR) that will detect V. parahaemolyticus were established. Consequently, two PCR primers could simultaneously amplify 285 bp and 368bp gene fragments from chromosomal DNA of V. parahaemolyticus in one PCR reaction, and no cross reaction was detected in 4 other pathogenic Vibrio species tested. The results of sensitivity of dulplex PCR showed that the two primers could detect V. parahaemolyticus at a level of as few as 8.8672×103CFU/mL, and detect purified chromosomal DNA at a level of as few as 0.0293 mg/L . Dominant V. parahaemolyticus could be isolated from the positive sample of dulplex PCR through detecting mysis, aquatic products and tank water samples. The PCR protocol amplifying gyrB and toxR gene fragments of the V. parahaemolyticus was established and could be useful in the specific and rapid diagnose of the disease caused by V. parahaemolyticus. |
| |
| Keywords: | Vibrio parahaemolyticus gyrB gene toxR gene dulplex PCR |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《海洋科学》浏览原始摘要信息 |
| 点击此处可从《海洋科学》下载免费的PDF全文 |
|
