Identification of immunodominant VP1 linear epitope of enterovirus71 (EV71) using synthetic peptides for detecting human anti-EV71 IgG antibodies in western blots

Abstract

A major IgG-specific immunodominant VP1 linear epitope of enterovirus71 (EV71) strain41 (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 Echo6. © 2008 European Society of Clinical Microbiology and Infectious Diseases.