Favorov M O, Khudyakov Y E, Mast E E, Yashina T L, Shapiro C N, Khudyakova N S, Jue D L, Onischenko G G, Margolis H S, Fields H A
Hepatitis Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Med Virol. 1996 Sep;50(1):50-8. doi: 10.1002/(SICI)1096-9071(199609)50:1<50::AID-JMV10>3.0.CO;2-1.
To develop an enzyme immunoassay (EIA) for IgM antibody to hepatitis E virus (HEV) (IgM anti-HEV) and IgG antibody to HEV (IgG anti-HEV), a synthetic gene encoding several liner immunodominant antigenic epitopes from HEV structural proteins was assembled as a chimeric recombinant mosaic protein (Mpr) with glutathione S-transferase and used as an immunodiagnostic target. In addition, a neutralization confirmation test was developed using individual synthetic peptides. Among 614 patients with acute hepatitis from 10 geographically distinct outbreaks, IgG anti-HEV was found in 546 (88.9%), with a range of 77-100% depending on the outbreak. Of 130 patients tested for IgM anti-HEV, 126 (96.9%) were positive. Among patients tested within 4 months of onset of jaundice, 37/37 (100%) were IgG anti-HEV positive. For patients from whom sera were collected 1-16 days after onset of jaundice, the geometric mean IgG titer (GMT) was 1:47,000; the GMT increased to 1:70,710 30-40 days after onset of jaundice and decreased to 1:1,778 3-4 months after the onset of jaundice. For patients tested 6-8 months after onset of jaundice, 11/12 (92%) were IgG anti-HEV positive, and the GMT was 1:2,908. IgM anti-HEV was detected in 43/43 (100%) sera collected 1-40 days after onset of jaundice, and the GMT for IgM anti-HEV was 1:10,000 at that time. For sera collected 3-4 and 6-12 months after onset of jaundice, 7/14 (50%) and 5/12 (40%) respectively, were IgM anti-HEV positive. In conclusion, an artificial mosaic protein composed of linear antigenic epitopes from open reading frame 2 (ORF2) and ORF3 of HEV has been successfully applied to the development of a sensitive and specific EIA for the detection of IgG and IgM anti-HEV activity. These assays were used for the verification of HEV infection in outbreak settings and for the diagnosis of HEV infection in sporadic cases.
为开发一种检测戊型肝炎病毒（HEV）IgM抗体（抗HEV IgM）和抗HEV IgG抗体的酶免疫测定法（EIA），将编码来自HEV结构蛋白的多个线性免疫显性抗原表位的合成基因组装成与谷胱甘肽S-转移酶的嵌合重组嵌合蛋白（Mpr），并用作免疫诊断靶标。此外，还使用单个合成肽开发了中和确认试验。在来自10个地理上不同疫情爆发地区的614例急性肝炎患者中，546例（88.9%）检测到抗HEV IgG，根据疫情爆发地区不同，比例范围为77%-100%。在130例检测抗HEV IgM的患者中，126例（96.9%）呈阳性。在黄疸发作后4个月内检测的患者中，37/37例（100%）抗HEV IgG呈阳性。对于黄疸发作后1-16天采集血清的患者，抗HEV IgG几何平均滴度（GMT）为1:47,000；黄疸发作后30-40天，GMT升至1:70,710，黄疸发作后3-4个月降至1:1,778。对于黄疸发作后6-8个月检测的患者，11/12例（92%）抗HEV IgG呈阳性，GMT为1:2,908。在黄疸发作后1-40天采集的43/43例（100%）血清中检测到抗HEV IgM，此时抗HEV IgM的GMT为1:10,000。对于黄疸发作后3-4个月和6-12个月采集的血清，抗HEV IgM阳性率分别为7/14例（50%）和5/12例（40%）。总之，由HEV开放阅读框2（ORF2）和ORF3的线性抗原表位组成的人工嵌合蛋白已成功应用于开发一种灵敏且特异的EIA，用于检测抗HEV IgG和抗HEV IgM活性。这些测定法用于疫情爆发情况下HEV感染的验证以及散发病例中HEV感染的诊断。
