关键词: 黄海希瓦氏菌;双抗体夹心ELISA;检测

Abstract:

At present, skin ulceration syndrome is the most serious disease in Apostichopus japonicus aquaculture. It caused mass mortality and great economic losses for aquaculture in China. A doubleantibody sandwich enzymelinked immunosorbent assay (DAS-ELISA) was developed for rapidly detecting S.smarflavi (AP629), a pathogen of Apostichopus japonicus, using polyclonal antibody (pAb) from rabbit and monoclonal antibody 3D9 (mAb 3D9) from mouse against AP629. The optimal dilution of pAb and mAb 3D9 were 1∶400 and 1∶80, respectively. This method has strong specificity, no cross reaction with other bacteria, including Vibrio sp, Aeromonas salmonicida, Edwardsiella ictaluri, and Escherichia coli etc., and high sensitivity in detection. The lowest concentration of strain AP629 that can be detected was 104 cells/mL and 106 cells/mL, respectively using PBS and homogenate of body wall from A. japonicus as medium. 10 artificial infected A.japonicus samples were detected and 100% of them were positive. So it has better stability and repetition. This method is very helpful for rapid and accurate diagnosis of A. japonicus infected by S.smarflavi AP629.

Key words: Shewanella smarflavi, double-antibody sandwich ELlSA, detection