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

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