一种用于检测戊型肝炎病毒假定中和抗体的酶联免疫吸附测定法可检测出针对1、2、3和4型的抗体。

An ELISA for putative neutralizing antibodies to hepatitis E virus detects antibodies to genotypes 1, 2, 3, and 4.

作者信息

Zhou Yi-Hua, Purcell Robert H, Emerson Suzanne U

机构信息

Hepatitis Viruses and Molecular Hepatitis Sections, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 50 South Drive MSC-8009, Bethesda, MD 20892, USA.

出版信息

Vaccine. 2004 Jun 30;22(20):2578-85. doi: 10.1016/j.vaccine.2003.12.017.

DOI:10.1016/j.vaccine.2003.12.017

PMID:15193383

Abstract

Two monoclonal antibodies that neutralize hepatitis E virus (HEV) were used to identify a subregion of ORF2 capsid protein spanning amino acids 459-607 as the shortest peptide to form the corresponding neutralization epitopes. An enzyme-linked immunosorbent assay (ELISA) based on a purified recombinant protein covering amino acids 458-607 in ORF2 of the Sar-55 strain (genotype 1) efficiently detected anti-HEV in non-human primates which had been experimentally infected with the four known mammalian genotypes of HEV, respectively. However, anti-HEV in these animals did not react with a shorter ORF2 peptide spanning amino acids 475-607. The ELISA was highly specific and sensitive when human or non-human primate sera were tested in parallel with a previously established ELISA based on amino acids 112-607 in ORF2. The antibody titer to peptides 458-607 in two ORF2-vaccinated rhesus monkeys which had different HEV challenge outcomes differed at the time of challenge. Since the ELISA appeared to be specific for neutralizing antibodies against HEV, it should be especially useful for quantifying the humoral immune response in hepatitis E vaccine trials.

摘要

使用两种中和戊型肝炎病毒（HEV）的单克隆抗体鉴定出ORF2衣壳蛋白中跨越氨基酸459 - 607的一个亚区域，作为形成相应中和表位的最短肽段。基于Sar - 55株（基因型1）ORF2中覆盖氨基酸458 - 607的纯化重组蛋白建立的酶联免疫吸附测定（ELISA），能有效检测分别经四种已知哺乳动物基因型HEV实验感染的非人灵长类动物体内的抗HEV抗体。然而，这些动物体内的抗HEV抗体与跨越氨基酸475 - 607的较短ORF2肽段无反应。当用人或非人灵长类动物血清与先前基于ORF2中氨基酸112 - 607建立的ELISA平行检测时，该ELISA具有高度特异性和敏感性。两只接种ORF2疫苗且对HEV攻击有不同结果的恒河猴，在攻击时针对肽段458 - 607的抗体滴度不同。由于该ELISA似乎对针对HEV的中和抗体具有特异性，它在戊型肝炎疫苗试验中对定量体液免疫反应应特别有用。