【摘要】 以差速离心法结合蔗糖密度梯度离心法提纯的PRRSVSC2株作为包被抗原,建立了检测猪生殖与呼吸综合征病毒(PRRSV)抗体的间接ELISA法。该法的最适抗原包被浓度为5μg/mL,最佳酶标兔抗猪抗体稀释度为1∶5000,对猪抗PRRSV的IgG检测灵敏度达到0.1μg/mL。该法只与PRRS阳性猪血清呈现阳性反应,而与PRRS阴性猪血清和猪伪狂犬病、猪乙型脑炎、猪瘟、猪细小病毒病阳性血清呈现阴性反应。应用该法检测PRRSVORF5基因疫苗免疫猪后的抗体消长规律表明,疫苗免疫猪后第7d抗体水平迅速上升,第75d达到高峰,然后缓慢下降,至第135d时仍然极显著(P<0.01)高于空载体对照和空白对照。试验结果表明,该法具有良好的特异性和敏感性,可用于PRRS流行病学调查和PRRSV基因疫苗免疫后抗体水平检测。更多还原

【Abstract】 An indirect ELISA for detection of antibodies againstporcine reproductive and respiratory syndrome virus (PRRSV) was established using PRRSV-SC2 isolate as coating antigen. The most suitable concentration of coating antigen was 5μg/mL, the most suitable dilution of enzyme labeledrabbit-anti-porcine antibody was 1∶5000, and the sensitivity of this method reached 0.1μg/mL for detection of IgG anti-PRRSV. This method showed a positive reaction only with PRRSV positive serum and did not react with positive serum with porcine pseudorabies, porcine epidemic encephalitis, swine fever and porcine parvovirus infection.Some serum samples from pigs immunized with ORF5 gene vaccine against PRRS were assayed. The result showed that antibody raised rapidly from day 7 post-immunization (PI), peaked on day 75 PI, then the antibody level reduced gradually, but still kept significant higher level compared with control group(P<0.01) on day 135 PI. The results showed that the method is specific, sensitive and stable, can be used as an ideal method in the epidemiological investigation and antibody test of pigs immunized with ORF5 gene vaccine against PRRS.更多还原