猪塞内卡病毒A型间接ELISA抗体检测方法的建立

Development of An ELISA Based on Synthetic Peptides for Detection of Antibodies Against Swine Senecavirus A

为了建立一种基于固相合成多肽技术的猪塞内卡病毒A型抗体血清学检测方法。本研究对猪塞内卡病毒A型VP1、VP2和VP3三种重要结构蛋白的B细胞表位进行设计并筛选到SVA1601、SVA1602、SVA1603和SVA1605共四条表位多肽,进而建立了一种用于检测猪塞内卡病毒A型抗体的间接ELISA方法,并验证该方法的检测敏感性、特异性、重复性以及与血清中和试验的符合率。试验结果显示,建立的间接ELISA方法对6份感染血清的最高稀释倍数为1:640,特异性为99.1%,批内和批间变异系数分别为2.4%~6.5%和4.4%~10.3%。与血清中和试验比较,符合率为97.7%。结果显示采用SVA1601、SVA1602、SVA1603和SVA1605建立的间接ELISA方法的敏感性较高,且重复性较好,可用于临床血清中猪塞内卡病毒A型特异性抗体的检测。

To establish a serological method based on synthetic-peptide technology for detection of antibodies against Senecavirus A(SVA).Four synthetic peptides(SVA1601,SVA1602,SVA1603 and SVA1605)based on three important structural proteins of SVA(VP1,VP2 and VP3)were screened and then used as coating antigens to establish an indirect enzyme-linked immunosorbent assay(ELISA)method for detecting SVA antibody.An ELISA system was optimized and the sensitivity,specificity,repeatability and coincidence rate of the method with serum neutralization test were studied.Sensitivity test results showed that the maximum dilution ratio of 6 positive sera was 1:640.The specificity was 99.1%.The Variations in intra-and inter-assay repeatability was 2.4%-6.5%and 4.4%-10.3%,respectively.Comparing this method with the serum neutralization test showed a coincidence rate of 97.7%.Results showed that our method has high sensitivity,good stability,and could be used to detect serum antibodies against SVA.