Detection of HEV antigen as a novel marker for the diagnosis of hepatitis E
Detection of HEV antigen as a novel marker for the diagnosis of hepatitis E
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DOI：
10.1002/jmv.20717
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发表时间：
2006-11-01
影响因子：
12.7
通讯作者：
Wang, Youchun
中科院分区：
文献类型：
--
作者：
Zhang, Feng;Li, Xiuhua;Wang, Youchun
戊型肝炎病毒(HEV)感染可以通过HEV RNA或抗HEV抗体的存在来诊断。建立了检测抗原的酶免疫分析(EIA)方法。制备了24株单抗。以三株单抗为包被抗体，建立检测HEV抗原的间接夹心EIA方法。抗-HEV-Ig M、抗-HEV-Ig M、抗-HEV-Ig G、抗-HEV-Ig G同时阳性的血清中，HEV抗原的阳性率分别为44.6%(33/74)、28.6%(50/175)和0(0/27)。对42份HEV抗体阳性血清进行HEV RNA和抗原平行检测，符合率为81.0%(34/42)。所有扩增产物均为HEV 4型。为探讨HEV抗原阳性与HEVRNA、抗HEVIg G、IgM及ALT浓度之间的时间关系，用HEV1、4型感染猕猴，采集连续标本。结果表明，EIA抗原可检测到两种基因型的衣壳蛋白。在ALT升高和抗HEV抗体出现之前，在感染的猴子中可检测到HEV抗原，并在所有病例中持续数周。HEV抗原在血清中的检测时间几乎与HEV RNA在粪便中的检测时间相同，但持续时间比HEV RNA短4周。这项检测对急性戊型肝炎的诊断很有价值，尤其是在抗-HEV血清转换前的窗口期。
Infection with hepatitis E virus (HEV) may be diagnosed by the presence of HEV RNA or antiHEV antibodies. An enzyme immunoassay (EIA) was developed for the detection of antigen. Twenty-four monoclonal antibodies (mAbs) were produced. An indirect sandwich EIA was developed to detect HEV antigen using a combination of three mAbs as coating antibodies. Approximately 44.6% (33/74), 28.6% (50/175), and none (0/27) of sera positive for anti-HEV IgM alone, both anti-HEV IgM and IgG, and anti-HEV IgG alone also were positive for HEV antigen using this EIA. Forty-two HEV antibody-positive sera were tested for HEV RNA and antigen in parallel and the concordance was 81.0% (34/42). All PCR products were found to belong to HEV genotype 4. In order to evaluate the temporal relationship between HEV antigen positivity and HEV RNA, anti-HEV IgG and IgM, and ALT concentrations, macaques were infected with HEV genotypes 1 and 4 and serial samples were collected. The results showed that the antigen EIA can detect the capsid proteins of both genotypes. HEV antigen was detectable prior to ALT elevation and the appearance of anti-HEV antibodies in the infected monkeys and lasted for several weeks in all cases. HEV antigen became detectable in the serum at almost the same time as HEV RNA in feces but persisted for 4 weeks less than HEV RNA. This assay should be valuable for the diagnosis of acute hepatitis E, particularly in the window period prior to seroconversion to anti-HEV.