双抗体夹心ELISA法检测养殖对虾病毒的研究

THE DETECTION OF THE VIRUSES IN CULTURED PENAEID SHRIBY DOUBLE ANTIBODY SANDWICH ELISA

用纯化的长毛对虾球状病毒（PPSV）和日本对虾中肠腺坏死杆状病毒（BMNV）制备新西兰兔抗BMNV和抗PPSV抗血清及Balb／c小鼠抗BMNV和抗PPSV抗血清，建立检测PSV和BMNV的双抗体夹心ELISA检测法，结果表明，双抗体夹心ELISA法具有较高的灵敏度，可以从100μl待测组织匀浆液中检测到50ng的PPSV蛋白，以及100ng的BMNV蛋白。不同病毒抗血清无交叉反应性，用该ELISA技术检测养殖对虾和多种采自养殖虾池及其附近的近海岸生物，发现相当比例的外观正常的对虾和近海岸生物已呈阳性反应，电镜观察结果证实了检测的特异性及可靠性。

The anhsera against P. penicillatus spheroid--virus(PPSV) and baculovilal nddgut glandnecrosis virus (BMN'V) have been obtained from rabbitS and face by imlnedzillg the aamalsrespechvely with Purified viruses and a double anhbody sandwich ELISA procedure has beenestablihed to detect PPSV and BMNV from culled shrimp and from beachy aamals sUrroudingthe culal farms. The resultS showed that the double anhbody sandwich ELISA was high sensihveand 50ng Protein of PPSV and 100ng protein of BMN'V could be detected from 100UI ofhepatopancreatic homogenate. Vilal detechon in the cultUred P, penicillatUs and P. japocicus andbeachy kilnals indicated that a high rate of the cultUred shrimp and beachy aamals with normalappeaxance Proedy have been infected by viruses. The high specifity and reliability of the detechonwere confirmed by the observahons of TEM.