title: Identification of specific antigenic epitope at N-terminal segment of enterovirus 71 (EV-71) VP1 protein and characterization of its use in recombinant form for early diagnosis of EV-71 infection.
abstract: Human enterovirus 71 (EV-71) is the main etiologic agent of hand, foot and mouth disease (HFMD). We sought to identify EV-71 specific antigens and develop serologic assays for acute-phase EV-71 infection. A series of truncated proteins within the N-terminal 100 amino acids (aa) of EV-71 VP1 was expressed in Escherichia coli. Western blot (WB) analysis showed that positions around 11-21 aa contain EV-71-specific antigenic sites, whereas positions 1-5 and 51-100 contain epitopes shared with human coxsackievirus A16 (CV-A16) and human echovirus 6 (E-6). The N-terminal truncated protein of VP1, VP₁₆₋₄₃, exhibited good stability and was recognized by anti-EV-71 specific rabbit sera. Alignment analysis showed that VP₁₆₋₄₃ is highly conserved among EV-71 strains from different genotypes but was heterologous among other enteroviruses. When the GST-VP₁₆₋₄₃ fusion protein was incorporated as antibody-capture agent in a WB assay and an ELISA for detecting anti-EV-71 IgM in human sera, sensitivities of 91.7% and 77.8% were achieved, respectively, with 100% specificity for both. The characterized EV-71 VP1 protein truncated to positions 6-43 aa has potential as an antigen for detection of anti-EV-71 IgM for early diagnosis of EV-71 infection in a WB format.
authors: Department of Microbiology and Immunology, School of Medicine, Southeast University, Nanjing, Jiangsu, PR China; Department of Microbiology and Immunology, School of Medicine, Shaoxing University, Shaoxing, Zhejiang, PR China.
authors: Jianhua
authors: J
authors: Zhang
authors: Department of Microbiology and Immunology, School of Medicine, Southeast University, Nanjing, Jiangsu, PR China.
authors: Bingfu
authors: B
authors: Jiang
authors: Department of Microbiology and Immunology, School of Medicine, Southeast University, Nanjing, Jiangsu, PR China.
authors: Mingjie
authors: M
authors: Xu
authors: Department of Dermatology, Affiliated Zhongda Hospital, School of Medicine, Southeast University, Nanjing, Jiangsu, PR China.
authors: Xing
authors: X
authors: Dai
authors: Division of Viral Hepatitis, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors: Michael A
authors: MA
authors: Purdy
authors: Department of Microbiology and Immunology, School of Medicine, Southeast University, Nanjing, Jiangsu, PR China. Electronic address: jihongmeng@163.com.
authors: Jihong
authors: J
authors: Meng
mesh_terms: D000328:Adult; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002675:Child, Preschool; D017124:Conserved Sequence; D042241:Early Diagnosis; D029821:Enterovirus A, Human / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006232:Hand, Foot and Mouth Disease / Q000175:diagnosis*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007223:Infant; D008297:Male; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D015678:Viral Structural Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D018985:Epitopes, B-Lymphocyte; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011994:Recombinant Proteins; D015678:Viral Structural Proteins
keywords: Early diagnosis; Enterovirus 71; Hand, Foot and mouth disease; Viral capsid protein