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学科主题基础医学
Application of Truncated Immunodominant Polypeptide from Hepatitis E Virus (HEV) ORF2 in an Assay To Exclude Nonspecific Binding in Detecting Anti-HEV Immunoglobulin M
Pan, Jin-Shun1,4; Zhang, Kui1; Zhou, Jinyong1,4; Wu, Chao2; Zhuang, Hui3; Zhou, Yi-Hua1,2,4
刊名JOURNAL OF CLINICAL MICROBIOLOGY
2010-03-01
DOI10.1128/JCM.01671-09
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
WOS记录号WOS:000274996200015
项目编号RC2007005 ; BK2009032 ; 30872235
资助机构Department of Health, Jiangsu Province, China ; Natural Science Foundation of Jiangsu Province, China ; National Natural Science Foundation of China
引用统计
被引频次:9[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/56205
专题北京大学基础医学院_病原生物学系
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
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GB/T 7714
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.
APA Pan, Jin-Shun,Zhang, Kui,Zhou, Jinyong,Wu, Chao,Zhuang, Hui,&Zhou, Yi-Hua.(2010).Application of Truncated Immunodominant Polypeptide from Hepatitis E Virus (HEV) ORF2 in an Assay To Exclude Nonspecific Binding in Detecting Anti-HEV Immunoglobulin M.JOURNAL OF CLINICAL MICROBIOLOGY,48(3),779-784.
MLA Pan, Jin-Shun,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".JOURNAL OF CLINICAL MICROBIOLOGY 48.3(2010):779-784.
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