Two kinds of hyperimmune sera,porcine-anti-rotavirus IgG and rabbit-anti-rotavirus IgG,were prepared by inoculating respectively piglets and rabbits with porcine rotavirus(RV) purified by diffe-rential centrifugation,and purified by affinity chromatography.A double antibody sandwich ELISA for detection of RV was developed based on the two kinds of IgG.The optimal coating concentration of porcine-anti-rotavirus IgG was 4 μg/mL,the optimal working concentration of rabbit-anti-rotavirus IgG was 3.5 μg/mL,the reaction time of sample was 90 min,and the optimal working dilution of HRP-labelled goat-anti-rabbit IgG was 1∶8 000.The positive standard value was 0.161(D 450 nm).The coefficient of variation of reproducibility was less than 10%,and at least 1.25 μg/mL antigen could be detectable.The ELISA had no cross-reaction with classical swine fever virus,porcine pseudorabies virus,porcine transmissible gastroenteritis virus,porcine epidemic diarrhea virus,Escherichia coli and Salmonella.The prepared plates and reagents could preserved at least four months at room temperature and 4 ℃.Seventy clinical fecal samples were detected by the ELISA and the colloidal gold card,and the positive ratio was 22.9% and 20.0% respectively.The results revealed that the ELISA possessed good specificity and reproducibility,and higher sensitivity,indicating a suitable method for rapid detection of RV.

轮状病毒(Rotavirus,RV)是一种重要的人兽共患病原,引起各种幼龄动物腹泻,在世界范围内造成严重的经济损失.RV属呼肠病毒(Reoviridae)科,轮状病毒属,为双链RNA病毒,其基因组不连续,由11个节段(Segment)的双链RNA组成,每个节段编码一种蛋白质,分为结构蛋白和非结构蛋白.