猪瘟病毒间接免疫荧光检测方法的建立

Establishment of an indirect immunofluorescence assay for detection of Classical swine fever virus

为了建立猪瘟病毒(CSFV)检测的间接免疫荧光(IFA)法,试验以猪瘟阳性血清为一抗、荧光素FITC标记的羊抗猪IgG为二抗,将CSFV接种于猪血管内皮细胞(EC)后,并对反应条件进行优化。结果表明:IFA法最佳工作条件为猪瘟阳性血清1∶200倍稀释,荧光素FITC标记的羊抗猪IgG荧光抗体1∶50倍稀释;用建立的IFA法检测CSFV感染的EC为阳性,而用猪圆环病毒(PCV)和猪传染性胃肠炎病毒(TGEV)感染的EC检测结果为阴性。说明建立的检测细胞培养中CSFV抗原的IFA法具有敏感性、特异性、简便快速等优点,可用于CSFV在EC上的感染和定位研究。

To establish an indirect assay （IFA） for detection Classical swine fever virus （ CSFV）, the positive serum of classical swine fever was used as primary antibody and an anti - fluorescein FITC - conjugated goat anti - pig IgG was used as secondary anti- body. CSFV was inoculated in the swine vascular endothelial cells （ EC）, the reaction conditions were optimized. The results showed that the optimal conditions for indirect immunofIuorescence assay in the porcine vascular endothelial cells were as follows： 1：200 -fold dilution of swine fever positive serum, and 1：50 - fold dilution of anti - fluorescein FITC - conjugated goat anti - pig IgG. It was found that the results were posi- tive in EC infected with CSFV, and were negative in EC infected with porcine circovirus （PCV） and transmissible gastroenteritis virus （TGE） using the IFA detection method established for the detections. The results indicate that this IFA test is a sensitive, specific and rapid method for the detection o f CSFV in the EC, which can be used for the research of CSFV infection and location on the EC.