戊型肝炎病毒IgG抗体酶联免疫诊断试剂盒研制

Development of a diagnostic kit of enzyme-linked immunoassay for detecting serum anti-hepatitis E virus IgG

目的 研制戊型肝炎病毒IgG抗体 (抗 HEVIgG)酶联免疫试剂盒 (ELISA)。方法 应用2个人工合成的HEV多肽抗原包被微量反应板 ,用兔抗人IgG (γ链 )辣根过氧化酶结合物作为第二抗体 ,检测血清中抗 HEVIgG ,建立抗 HEVIgGELISA ;对该试剂盒的灵敏度、特异度、精密度、稳定性进行测定和临床考核。结果 经中国药品生物制品检定所检定 ,本试剂盒的灵敏度为 90 % (1 10 ) ,特异度为 10 0 % (0 30 ) ,精密度 <15 % ,符合国家对抗 HEVIgGELISA的要求 ;在 4℃下的稳定性至少为 1年 ;与美国Genelabs公司和新加坡DBL抗 HEVIgGELISA总符合率分别为 10 0 % (4 3 43)和 96 6 %(86 89)。该试剂盒于 1998年通过国家药品监督管理局的新药评审 ,获国家新药证书。结论 抗 HEVIgGELISA可用于戊型肝炎病毒感染的流行病学调查和临床诊断。

Objective To develop a diagnostic kit for detecting serum anti-hepatitis E virus (HEV) IgG with enzyme-linked immunoassay (ELISA). Methods The diagnostic kit of detecting anti-HEV IgG with ELISA was prepared by two synthetic HEV peptides used for coating the solid-phase to capture the antibody against HEV in serum,and then by horseradish peroxidase antihuman IgG (γ chain) added as the second antibody to bind the anti-HEV IgG in serum. Its sensitivity,specificity,precision and stability were measured and clinically evaluated. Results The sensitivity,specificity,and precision of the diagnostic kit detected by the China National Institute for the Control of Pharmaceutical and Biological Products were 90% (1/10),100% (0/30) and <15%,respectively,meeting the national standards for detecting serum anti-HEV IgG with ELISA. The ELISA Kit is stable for 1 year at least under 4 ℃. The total consistency rates were 100% (43/43) and 96.6% (86/89),as compared with Genelabs and Singaporean DBL kits. This diagnostic kit for detecting serum anti-HEV IgG with ELISA was evaluated and approved by the State Drug Administration,China and a certificate was conferred in 1998. Conclusions The diagnostic kit for detecting anti-HEV IgG with ELISA can be used for epidemiological studies and clinical diagnosis of HEV infection.