摘要目的 建立猴痘病毒感染血清IgG抗体检测方法.方法 采用合成的猴痘病毒B21R蛋白上的2条混合多肽作为包被抗原,建立猴痘病毒感染人血清IgG抗体间接ELISA检测方法.选取正常成人血清、无关病毒感染血清和猴痘病毒感染者血清标本对多肽抗原特异性进行评价,并优化反应参数.结果 猴痘病毒B21R蛋白上的二肽混合抗原与正常血清和无关病毒感染血清均无明显交叉反应,具有较好的抗原特异性.经优化后,混合二肽抗原的最佳包被浓度均为50 ng/孔,二抗稀释度为1:20000.在此条件下塞拉利昂猴痘患者血清在12 d和55 d的猴痘病毒IgG抗体滴度均为1:6400.结论 初步建立了猴痘病毒感染血清IgG抗体间接ELISA检测方法,为我国应对可能出现的猴痘输入病例提供技术储备.

abstractsObjective To establish a method for detection of human antibodies against monkeypox virus. Mothds The enzyme linked immunosorbent assay (ELISA) plates were coasted with two monkeypox virus peptides from B21R protein, to establish an indirect ELISA for detecting monkeypox virus IgG antibody. The healthy individuals serum samples, monkeypox virus infected patient serum samples and other virus infected patient sera samples were applied to evaluate specificity of the peptides antigen. The reaction conditions were optimized. Results Synthesized two peptides from monkeypox virus BR21R protein did not cross react obviously with healthy person serum and other virus infected serum. It was shown that the reaction condition was best with sera dilution at 1: 50 when two combined peptides were coated at 100 ng / well, and second - antibody was diluted at 1: 20000. At this condition the cut off value of IgG antibody in serum samples for ELISA were A450 reading of 0. 393. The detected results of two serum samples collected from the monkeypox patient in Sierra Leone were strongly positive, the titers of IgG antibody in two sera were both 1: 6400. Conclusions The indirect ELISA for detection of monkeypox virus infection was established preliminarily which provided useful tools for epidemiological study and diagnosis.