检测鲆鱼腹水病病原菌迟缓爱德华氏菌和溶藻弧菌的嵌套PCR方法

DETECTION OF PATHOGENIC BACTERIAL (EDWARDSIELLA TARDA AND VIBRIO ALGINOLYTICUS) ASSOCIATED WITH SWOLLEN ABDOMEN OF CULTURED TURBOT (SCOPHTHALMUS MAXIMUS) AND FLOUNDER (PARALICHTHYS OLIVACEUS) BY NESTED-PCR

本文首次建立了用嵌套式聚合酶链式反应(Nested-PCR)分别检测天津地区海水工厂化养殖大菱鲆和褐牙鲆腹水病病原菌迟缓爱德华氏菌(Edwardsiella tarda)和溶藻弧菌(Vibrio alginolyticus)的方法。以E781F和E781R为引物,扩增迟缓爱德华氏菌溶血素基因中781bp的片段,再以E485F和E485R为引物,扩增其中的485bp的片段;以V899F和V899R为引物,扩增溶藻弧菌胶原酶基因中899bp片段,再以V550F和V550R为引物,扩增其中的550bp片段。以其他常见海水鱼类致病菌为阴性对照,结果均无非特异性扩增。对迟缓爱德华氏菌和溶藻弧菌的检出灵敏度分别可达10fg和1fg细菌DNA,显著优于目前的检测方法。作者应用该技术对天津地区海水工厂化大菱鲆和褐牙鲆于2006年3月至10月连续进行监测,为有效地控制该病的流行提供了技术支撑。

Turbot(Scophthatmus maximus) and flounder(Paralichthys olivaceus) are rare and economic fish in the north of China.In recent years,they have become the dominant cultured varieties.Tianjin is the main cultural area of turbot and flounder.However,the prevalence of diseases of the cultured turbot and flounder has resulted in great loss to the cultural enterprise.Swollen abdomen disease was the serious threat to the turbot and flounder according to the investigation.The mortality can reach 80%—90% while the dis...