间接免疫荧光检测猪瘟活疫苗效价方法的建立

Establishment of an indirect immunofluorescence assay for classical swine fever live vaccine(cell line origin)

为了寻找到猪瘟活疫苗(传代细胞源)效力检验的实验室替代方法,本研究将检验合格的疫苗按头份用专用稀释液10倍梯度稀释后,接种至长满猪睾丸细胞(ST细胞)单层的细胞板上,以猪瘟阳性血清为一抗、山羊抗猪IgG(FITC标记)为二抗,通过反应体系条件的优化,确定病毒接种细胞后培养72 h,一抗和二抗的稀释度为1∶400倍,建立了猪瘟活疫苗(传代细胞源)成品苗的间接免疫荧光(Indirect immunofluorescence assay,IFA)检测方法;此方法敏感性、特异性和重复性良好。用建立的IFA方法检测10批次猪瘟活疫苗(传代细胞源)试验苗,结果为:104.67~105.33TCID50/头份。该方法的建立可为猪瘟活疫苗(传代细胞源)的质量控制提供技术支持和检验参考。

In order to find an alternative laboratory method for the efficacy test of classical swine fever live vaccine(cell line origin),in this study the qualified vaccine was 10 times gradiently diluted with special diluent by doses and inoculated into the cell plate covered with swine testicles cells and cultured for 72 h. The dilution of the positive serum of classical swine fever virus(CSFV)and the goat anti-pig IgG(FITC marker)was determined as 1∶400 both. A method of indirect immunofluorescence assay(IFA)was established for detection of classical swine fever live vaccine(cell line origin),this method has good sensitivity,specificity and reproducibility. The IFA method was used to detect10 batches of classical swine fever live vaccine(cell line origin),the results were 104.67~105.33TCID50/dose. The established IFA detection method can provide technical support to quality control of classical swine fever live vaccine(cell line origin).