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

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 i...

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Bibliographic Details
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
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Summary:	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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1198-743X 1469-0691
DOI:	10.1111/j.1469-0691.2007.01904.x