仿刺参及养殖环境中溶藻弧菌和灿烂弧菌的PCR快速检测

doi:10.3969/j.issn.1008-0864.2010.03.22

›› 2010, Vol. 12 ›› Issue (3): 125-130.DOI: 10.3969/j.issn.1008-0864.2010.03.22

仿刺参及养殖环境中溶藻弧菌和灿烂弧菌的PCR快速检测

汪笑宇,周遵春,关晓燕,姜北,陈仲,董颖,杨爱馥

(辽宁省海洋水产科学研究院, 辽宁省海洋水产分子生物学重点实验室, 辽宁大连 116023)

收稿日期:2010-01-06 修回日期:2010-02-23 出版日期:2010-06-15 发布日期:2010-04-02
通讯作者: 周遵春,研究员,博士,研究方向为海洋生物技术。E-mail:zunchunz@hotmail.com
作者简介:汪笑宇,硕士,研究方向为海洋生物病害。E-mail:smallouc@163.com
基金资助:
国家海洋公益项目（200705007）资助。

Rapid PCR Detection for Vibrio alginolyticus and Vibrio splendidus in Sea Cucumber Apostichopus japonicas and its Culturing Environment

WANG Xiao-Yu, ZHOU Zun-Chun, GUAN Xiao-yan, JIANG Bei, CHEN Zhong, DONG Ying, YANG Ai-fu

(Liaoning Key Laboratory of Marine Fishery Molecular Biology, Liaoning Ocean and Fisheries Science Research Institute, Liaoning Dalian 116023, China)

Received:2010-01-06 Revised:2010-02-23 Online:2010-06-15 Published:2010-04-02

摘要/Abstract

摘要：

溶藻弧菌和灿烂弧菌是水产养殖中常见的致病菌，给水产动物的养殖带来巨大的损失，建立一种简便、快速、有效的检测方法十分必要。根据溶藻弧菌和灿烂弧菌gyrB基因序列的保守区序列分别设计引物，对9株溶藻弧菌和6株灿烂弧菌进行了PCR扩增。结果表明两对引物能特异地检测溶藻弧菌和灿烂弧菌，而与其他细菌没有交叉反应；检测溶藻弧菌的每个PCR反应的敏感度为0.13 pg的DNA和103 cfu/mL细菌，检测灿烂弧菌的每个PCR反应的敏感度为0.34 pg的DNA和103 cfu/mL细菌。该PCR检测方法具有特异性好、灵敏度高的特点，可用于对感染溶藻弧菌和灿烂弧菌的仿刺参及其养殖水体中的细菌性病原进行快速检测。

关键词: 溶藻弧菌;灿烂弧菌;gyrB基因;PCR;快速检测

Abstract:

Vibrio alginolyticus and Vibrio splendidus strains are common bacterial pathogens, having a significant negative economic impact on aquaculture. It is necessary to establish asimple, rapid and effective detection method. According to the conserved sequence of gyrB gene of V. alginolyticus and V. splendidus, 2 pairs of primers were designed respectively to amplify 9 strains of V. alginolyticus and 6 strains V. splendidus. The results showed that the two pairs of primers could specifically detect V. alginolyticus and V. splendidus, but there is no specific band with bacteria which exist extensively in the culture of the sea cucumber. The minimum detectable amount of template was 0.13 pg/μL DNA or the lysed products of 103 cfu/mL of V. alginolyticus, and 0.34 pg/μL DNA or the lysed products of 103 cfu/mL of V. splendidus. This method had good specificity and high sensitivity. It was able to rapidly detect V. alginolyticus and V. splendidus in sea cucumber and its culturing environment.

Key words: Vibrio alginolyticus, Vibrio splendidus, gyrB, PCR, rapid detection

中图分类号:

S947.9

汪笑宇,周遵春,关晓燕,姜北,陈仲,董颖,杨爱馥. 仿刺参及养殖环境中溶藻弧菌和灿烂弧菌的PCR快速检测[J]. , 2010, 12(3): 125-130.

WANG Xiao-Yu, ZHOU Zun-Chun, GUAN Xiao-yan, JIANG Bei, CHEN Zhong, DONG Ying, YA. Rapid PCR Detection for Vibrio alginolyticus and Vibrio splendidus in Sea Cucumber Apostichopus japonicas and its Culturing Environment[J]. , 2010, 12(3): 125-130.

仿刺参及养殖环境中溶藻弧菌和灿烂弧菌的PCR快速检测

Rapid PCR Detection for Vibrio alginolyticus and Vibrio splendidus in Sea Cucumber Apostichopus japonicas and its Culturing Environment

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