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学科主题: 基础医学
题名:
Application of Truncated Immunodominant Polypeptide from Hepatitis E Virus (HEV) ORF2 in an Assay To Exclude Nonspecific Binding in Detecting Anti-HEV Immunoglobulin M
作者: Pan, Jin-Shun1,4; Zhang, Kui1; Zhou, Jinyong1,4; Wu, Chao2; Zhuang, Hui3; Zhou, Yi-Hua1,2,4
刊名: JOURNAL OF CLINICAL MICROBIOLOGY
发表日期: 2010-03-01
DOI: 10.1128/JCM.01671-09
卷: 48, 期:3, 页:779-784
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Microbiology
研究领域[WOS]: Microbiology
关键词[WOS]: LINKED-IMMUNOSORBENT-ASSAY ; ANTIBODIES ; INFECTION ; GENOTYPE ; PROTEIN ; IDENTIFICATION ; VACCINE ; DIVERSITY ; DIAGNOSIS ; SEQUENCE
英文摘要:

The diagnosis of recent hepatitis E virus (HEV) infection depends on serologic testing for anti-HEV IgM; however, false-positive results may occur. In the present study, we cloned the ORF2 fragment of genotype 4 HEV and demonstrated that a subregion covering amino acids 459 to 607 in ORF2 forms the immunodominant B-cell epitopes, as it does in genotype 1 viruses. Truncation of several residues from either the N or C terminus of the polypeptide abolished the reactivity of anti-HEV from naturally infected persons. By the combination of high reactivity of the immunodominant polypeptide and poor reactivity of the truncated polypeptide, we established an indirect enzyme-linked immunosorbent assay (ELISA) to detect anti-HEV IgM. In this assay, all 37 sera that were HEV RNA positive reacted with the immunodominant polypeptide but not with the truncated one, and none of 159 sera from healthy persons reacted with either of the polypeptides. In retesting of 117 sera that originally tested positive for anti-HEV IgM, using a Genelabs kit, only 34 were positive and 83 were negative. Western blot analyses and other experiments strongly indicated that these 83 discordant sera were negative for anti-HEV IgM. Furthermore, among the 117 sera, 5 reacted with both the immunodominant and truncated polypeptides, with comparable optical densities at 450 nm. However, their reactivity was demonstrated to result from nonspecific binding. Together, the data indicate that the poor reactivity of a truncated ORF2 polypeptide can be used to exclude nonspecific binding in the detection of anti-HEV IgM.

语种: 英语
所属项目编号: RC2007005 ; BK2009032 ; 30872235
项目资助者: Department of Health, Jiangsu Province, China ; Natural Science Foundation of Jiangsu Province, China ; National Natural Science Foundation of China
WOS记录号: WOS:000274996200015
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/56205
Appears in Collections:基础医学院_病原生物学系_期刊论文

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作者单位: 1.Nanjing Univ, Sch Med, Nanjing Drum Tower Hosp, Dept Lab Med, Nanjing 210008, Peoples R China
2.Nanjing Univ, Sch Med, Nanjing Drum Tower Hosp, Dept Infect Dis, Nanjing 210008, Peoples R China
3.Jiangsu Key Lab Mol Med, Nanjing 210008, Peoples R China
4.Peking Univ, Hlth Sci Ctr, Dept Microbiol, Beijing 100191, Peoples R China

Recommended Citation:
Pan, Jin-Shun,Zhang, Kui,Zhou, Jinyong,et al. Application of Truncated Immunodominant Polypeptide from Hepatitis E Virus (HEV) ORF2 in an Assay To Exclude Nonspecific Binding in Detecting Anti-HEV Immunoglobulin M[J]. JOURNAL OF CLINICAL MICROBIOLOGY,2010,48(3):779-784.
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