Identification of immunodominant VP1 linear epitope of enterovirus 71 (EV71) using synthetic peptides for detecting human anti-EV71 IgG antibodies in western blots

• Published in: Clinical microbiology and infection Vol. 14; no. 3; pp. 286 - 288
• Main Authors: Foo, D.G.W., Ang, R.X., Alonso, S., Chow, V.T.K., Quak, S.H., Poh, C.L.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Elsevier Ltd 01-03-2008; Blackwell Publishing Ltd; Blackwell
• Subjects: Antibodies, Viral - blood; Antibodies, Viral - immunology; Antigens, Viral - immunology; Biological and medical sciences; Blotting, Western; Child; Cloning, Molecular; Detection; enterovirus 71; Enterovirus A, Human - immunology; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Humans; Immunodominant Epitopes - genetics; Immunodominant Epitopes - immunology; immunodominant linear epitope; Immunoglobulin G - blood; Immunoglobulin G - immunology; Microbiology; Oligonucleotides - genetics; pepscan analysis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - isolation & purification; recombinant GST-SP32 fusion protein; Sensitivity and Specificity; synthetic peptide; Techniques used in virology; Virology; pepscan analysis; recombinant GST-SP32 fusion protein; enterovirus 71; Detection; immunodominant linear epitope; synthetic peptide; Human; Enterovirus 71; Antigenic determinant; Peptides; Picornaviridae; Antibody; IgG; Enterovirus; Identification; Virus; Infection; Immunoblotting assay; Diagnosis; Recombinant protein; Fusion protein; Aminoacid sequence
• ISSN: 1198-743X; 1469-0691
• DOI: 10.1111/j.1469-0691.2007.01904.x

A major IgG-specific immunodominant VP1 linear epitope of enterovirus 71 (EV71) strain 41 (5865/SIN/00009), defined by the core sequence LEGTTNPNG, was identified by Pepscan analysis. Oligonucleotides corresponding to the amino-acid sequence of synthetic peptide SP32 were cloned and over-expressed in Escherichia coli as a recombinant glutathione-S-transferase (GST)–SP32 fusion protein. In ELISAs, this protein did not react with human anti-EV71 IgG antibodies, but there was significant immunoreactivity according to western blot analysis. The amino-acid sequence of SP32 was highly specific for detecting EV71 strains in western blot analysis, and showed no immunoreactivity with monoclonal antibodies raised against other enteroviruses, e.g., CA9 and Echo 6.