动物血清中SARS-CoV-2抗体检测方法的建立与评价

Establishment and Evaluation of A Method for Detecting SARS-CoV-2 Antibodies in Animal Sera

为针对不同动物开展SARS-CoV-2相关研究工作提供检测手段,针对SARS-CoV-2 S1蛋白建立了一种可检测不同动物血清中的SARS-CoV-2抗体的ELISA方法。通过构建表达SARS-CoV-2 S1蛋白的重组杆状病毒,收获重组杆状病毒培养上清纯化蛋白,建立SARS-CoV-2双抗原夹心ELISA抗体检测方法,并评价该方法的特异性、敏感性及临床适用性。结果显示,获得可表达SARS-CoV-2 S1蛋白的重组杆状病毒,表达纯化获得SARS-CoV-2 S1蛋白,并对该蛋白进行HRP标记,建立了SARS-CoV-2双抗原夹心ELISA抗体检测方法,该方法检测SPF小鼠、兔、猪及动物源性冠状病毒抗体阳性血清均为阴性;检测SARS-CoV-2 S1蛋白免疫兔血清,稀释至1∶320时仍为阳性;检测SARS-CoV-2人工感染雪貂灭活血清,感染前均为阴性,感染后13 d和20 d均为阳性。表明建立的SARS-CoV-2双抗原夹心ELISA抗体检测方法具有良好的特异性、敏感性和临床适用性,可用于多种动物血清中SARS-CoV-2抗体的检测。

In order to carry out the research on the detection of SARS-Cov-2 antibodies in different animals,an ELISA method was established to detect the antibodies of SARS-Cov-2 in different animal sera.By constructing the recombinant baculovirus expressing SARS-CoV-2 S1 protein,and purifing protein from harvesting supernatant.The detection antibody method of SARS-Cov-2 double antigen sandwich ELISA was established,and the specificity,sensitivity and clinical applicability of the method were tested.The results showed that the recombinant baculovirus expressing SARS-Cov-2 S1 protein and the purified SARS-Cov-2 S1 protein were obtained.The double antigen sandwich ELISA detected SPF mouse,rabbit,pig sera and ani-mal origin coronavirus positive sera were negative;detected rabbit serum immunized with S1 protein of SARS-Cov-2 remained positive until 1∶320 dilution;detected the inactivated serum of ferret artificially infected with SARS-Cov-2,sera before infection were negative,but positive at 13 and 20 days after infection.The double antigen sandwich ELISA for detection of SARS-Cov-2 antibody established in this study has good specificity,sensitivity and clinical applicability,and can be used for the detection of SARS-Cov-2 antibody in a variety of animal sera.