文蛤副溶血弧菌间接ELISA检测技术的研究  

Study on Indirect ELISA Method for Detecting Vibrio Parahaemolyticus in Hard Clam Meretrix Meretrix Linnaeus

作　　者：李国[1] 闫茂仓[2] 常维山[1] 刘连生[3] 马爱敏[1] 林志华[2] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]浙江省海洋水产养殖研究所,浙江温州325005 [3]安徽农业大学生命科学学院,安徽合肥230036

出　　处：《海洋通报》2008年第5期85-90,共6页Marine Science Bulletin

基　　金：国家科技支撑计划(2007BAD43B09);国家高技术研究发展计划(2006AA10A410);浙江省重大科技攻关计划(2006C12013)

摘　　要：从患病文蛤组织内分离出一株细菌,回归感染试验证明是文蛤的致病菌,经鉴定为副溶血弧菌,灭活后(56℃,1h)制成菌苗,用该菌苗制备兔抗血清,以辣根过氧化酶标记的羊抗兔血清(HRP-IgG)为酶标二抗,对副溶血弧菌进行间接ELISA快速检测。结果表明:应用间接ELISA技术检测副溶血弧菌具有较高的灵敏度,其最低检测极限为1×105cfu/mL。患病文蛤内脏团中副溶血弧菌的检出率为80%,无病症带菌文蛤中的检出率为15%,海水中副溶血弧菌的浓度低于最低检测极限。表明间接ELISA检测法不仅可以用于发病文蛤的快速检测,而且能够检测出无病症的带菌的文蛤,为有效预防副溶血弧菌病的暴发提供了检测手段。Pathogenic bacterium was isolated form diseased hard clam Meretrix meretrix Linnaeu, and it was identified to be Vibrio parahaemolyticus （Vp）, and was confirmed to be the pathogens of the epidemic by artificial infection experiment. This pathogen was inactivated （56℃, 1h）, to be as vaccine,the antiserum specific for Vp was obtained from rabbits by injecting vaccine; and the anti-rabbit IgG goat was marked by horseradish peroxidase （HRP）and used in an indirect ELISA detection. The results showed the lowest VP suspension detection limit in laboratory was 1×10^5cfu/mL on worksite. Vp were detected by ELISA form samples of diseased and healthy clam with 80 % and 15 % respectively, but none for the seawater. This indicated that ELISA could be used not only to diagnose the clinically diseased clam Meretrix meretrix, but also to recognize pathogen-carrier clam Meretrix meretrix Linnaeus, and it can provide basis to preclude the explosion of the vibriosis.

关 键 词：文蛤 副溶血弧菌 弧菌病 间接ELISA 快速检测 

分 类 号：Q178.53[生物学—水生生物学]