WSSV单抗的制备及其在红螯螯虾病毒病检测中的应用

将提纯的感染红螯螯虾的WSSV，免疫BALB／c小鼠，三次免疫后取其脾细胞与SP2／0融合．间接ELISA筛选，阳性克隆经3次亚克隆后，共获得7株针对WSSV的特异性单抗，分别命名为E2、C2、E3、G3、C4、D5以及F10．细胞上清ELISA效价为1：1600—1：6400．抗体亚类鉴定结果表明：E2、G3、C4属于IgG1，C2、D5、F10属于IgG3，E3属于IgG2a亚类；单抗热稳定性试验表明7株单抗均是热稳定的．选取单抗G3和F10进行病毒中和试验，结果表明：在适宜的抗原浓度下，两株单抗均有较好的中和能力．选取单抗G3作为一抗建立了间接ELISA方法，通过对人工感染红螯螯虾WSSV的测定，初步证实该单抗可用于WSSV的检测．

Preparation of Monoclonal Antibodies Against WSSV and Application to Test the Virus in Redclaw Crayfish

The 8-week-old female BALB/c mice were immunized with purified WSSV. Spleen cell collected from immunized mice were fused with SP2/0 myeloma cells after the third immunization. Positive hybri- doma ceils were screened by indirect ELISA and subeloned by limited dilution method. After three times of subeloning, seven McAbs against the WSSV were selected, and were designated as E2, C2, E3, G3, C4, D5 and F10 respectively. The seven McAbs were proved to have high ELISA titers between 1：1 600 - 1 ： 6 400. E2, G3 and CA were identified to be IgG1 ; C2, D5 and F10 to be IgG3 ; E3 to be IgG2a respectively. G3 and F10 were chosen to do neutralization test with WSSV. They were proved to have good neutralization character. Indirect ELISA was established to detect WSSV with G3 as first antibody. The result suggested that G3 might be used to detect WSSV in Redclaw Crayfish, and that it could have wide application prospects in shrimp production.