用间接免疫荧光检测方法评价猪瘟兔化弱毒活疫苗效力的研究

Using an Indirect Immunofluorescence Assay for Evaluating the Efficacy of Live Classical Swine Fever Virus Vaccine (C Strain)

关键词:  猪瘟疫苗  效力  兔体感染量  间接免疫荧光

Key words:  Classical swine fever vaccine  efficacy  rabbit infection dose  indirect immunofluorescence assay

摘要:

为用体外试验方法评价猪瘟兔化弱毒活疫苗效力,以猪瘟病毒(CSFV)抗体(兔源)为一抗、荧光素标记羊抗兔IgG 为二抗,建立了CSFV兔化弱毒株的间接免疫荧光检测方法(IFA)。特异性试验表明,用该IFA方法检测CSFV兔化弱毒株接种的RK细胞为阳性,而检测伪狂犬病毒(PRV)、猪细小病毒病(PPV)、牛病毒性腹泻/粘膜病毒(BVDV)接种的RK细胞均为阴性。CSFV兔化弱毒株和活疫苗的兔体感染量(RID)用兔体测定,半数组织感染量(TCID₅₀)用IFA测定,拟合RID与TCID₅₀的线性回归方程,确定1RID/ml=(TCID₅₀/0.1ml 200)/12。

Abstract:

Using the antibody against CSFV (C strain) as the primary antibody and the FITC labeled goat anti-rabbit IgG as the second antibody, an indirect immunofluorescence assay was developed to evaluate the efficacy of live classical swine fever virus vaccine (C strain). The IFA showed positive result in detection RK cells inoculated with CSFV (C strain), but negative results in detection RK cells inoculated with pseudorabies virus, porcine parvovirus and bovine viral diarrhea-mucosal virus. Rabbit infection dose (RID) tested with rabbits and 50% tissue culture infective dose (TCID₅₀) tested with IFA in CSFV (C strain) and vaccine were both carried out. The linear regression equation fitting the TCID₅₀over RID was conducted, and the result is that 1RID/ml= (TCID₅₀/0.1ml 200)/12.