人戊型肝炎病毒IgG、IgM及IgA抗体ELISA检测方法的建立与初步应用

ELISA detection method for IgG, IgM and IgA antibodies against hepatitis E virus: Establishment and preliminary application

目的建立用于检测戊型肝炎病毒(HEV)IgG、IgM及IgA抗体的间接ELISA方法。方法用重组HEV杆状病毒感染Tn-5细胞表达HEV病毒样颗粒(VLP),用作包被抗原,建立间接ELISA检测方法,进行方法学评估,分析云南省西双版纳地区献血者HEV抗体流行率。结果收集纯化的VLP能具有HEV抗原性。西双版纳州献血人群抗-HEV IgG、IgM和IgA阳性率分别为18%(90/500)、5.6%(28/500)和2.6%(13/500),HEV近期感染率为1.8%(9/500),戊型肝炎血清流行率为19.8%(99/500)。13例抗-HEV IgA阳性标本中,有2例抗-HEV IgG和IgM双阴性。结论云南省西双版纳自治州献血者HEV抗体流行率较高,部分献血者为近期感染,对输血安全可能构成威胁。HEV IgA抗体间接ELISA方法联合HEV IgM抗体检测,能提高HEV近期感染者检出率。

Objective To establish an indirect ELISA method for detecting IgG,IgM and IgA antibodies against Hepatitis E virus(HEV). Methods Tn-5 cells were infected with recombinant HEV baculovirus, and HEV-like particles(VLP) were collected and purified for coating antigen. The reaction conditions and methodology of indirect ELISA method were established and evaluated. The prevalence of HEV antibody among blood donors in Xishuangbanna were detected. Results The collected and purified VLP showed HEV antigenicity. The positive rates of anti-HEV IgG, IgM and IgA in blood donors in Xishuangbanna Prefecture were 18%(90/500), 5.6%(28/500) and 2.6%(13/500), respectively. The recent HEV infection rate was 1.8 %(9/500), and the seroprevalence of hepatitis E was 19.8%(99/500). Of the 13 anti-HEV IgA positive samples, 2 were both anti-HEV IgG and IgM negative. Conclusion HEV antibody positive is common among blood donors in Xishuangbanna, some of which are recent infections, posing a threat to the safety of blood transfusion. HEV IgA antibody indirect ELISA combined with human HEV IgM antibody detection can improve the detection rate of recent HEV infection.