非洲猪瘟病毒抗体量子点检测试纸条的研制

Establishment of a quantum dots-based immunochromatographic strip for detection of the antibodies against African swine fever virus

根据非洲猪瘟病毒(ASFV)VP73蛋白的氨基酸序列推测抗原表位优势区域,合成多肽并与牛血清白蛋白(BSA)偶联,筛选有抗原性的多肽作为试纸条检测线的包被抗原,采用量子点作为标记材料对葡萄球菌蛋白A(SPA)进行标记,以抗SPA的多克隆抗体作为质控线的包被蛋白,制备快速检测ASFV抗体的量子点免疫层析试纸条。结果表明,所制备的试纸条与常见相关猪疫病阳性血清无交叉反应,与进口ELISA试剂盒对临床样品的检测符合率为100%;特异性强、敏感性高、稳定性好、操作简便,可用于ASFV抗体的快速检测和流行病学调查。

According to the amino acid sequence of the African swine fever virus（ASFV） VP73 protein,the antigen epitope dominant region was predicted and compounded,and then conjugated with bovine serum albumin（BSA）.The polypeptide was immobilized on the detection zone of the immunochromatographic strip,and the rabbit anti-protein A was immobilized on the control zone.The water-soluble quantum dots（QDs） were used as the signal output and conjugated to streptococcal protein A（SPA）,which was capable of binding to immunoglobulin G（Ig G） from many species through an amide bond to capture the target anti-ASFV Ig G.The results showed that the immunochromatographic strip was specific for ASFV-positive sera.No cross-reactions were observed for the positive sera against other porcine viruses.The coincidence rate between the strip and a commercial ELISA kit was 100%.The strip is specific,sensitive,stable,and easy to operate.It can be used for rapid detection of anti-ASFV antibodies and epidemiological investigation.