Analysis of cross-reactive neutralizing antibodies in human HFMD serum with an EV71 pseudovirus-based assay

PLoS One. 2014 Jun 25;9(6):e100545. doi: 10.1371/journal.pone.0100545. eCollection 2014.


Hand, foot and mouth disease, associated with enterovirus 71 (EV71) infections, has recently become an important public health issue throughout the world. Serum neutralizing antibodies are major indicators of EV71 infection and protective immunity. However, the potential for cross-reactivity of neutralizing antibodies for different EV71 genotypes and subgenotypes is unclear. Here we measured the cross-reactive neutralizing antibody titers against EV71 of different genotypes or subgenotypes in sera collected from EV71-infected children and vaccine-inoculated children in a phase III clinical trial ( Identifier: NCT01636245) using a new pseudovirus-based neutralization assay. Antibodies induced by EV71-C4a were cross-reactive for different EV71 genotypes, demonstrating that C4a is a good candidate strain for an EV71 vaccine. Our study also demonstrated that this new assay is practical for analyses of clinical samples from epidemiological and vaccine studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / blood*
  • Antibodies, Neutralizing / immunology*
  • Child, Preschool
  • Cross Reactions*
  • Enterovirus A, Human / genetics
  • Enterovirus A, Human / physiology*
  • Genotype
  • HEK293 Cells
  • Hand, Foot and Mouth Disease / blood*
  • Hand, Foot and Mouth Disease / immunology
  • Hand, Foot and Mouth Disease / virology*
  • Humans
  • Luciferases, Firefly / genetics
  • Neutralization Tests*
  • Viral Vaccines / immunology


  • Antibodies, Neutralizing
  • Viral Vaccines
  • Luciferases, Firefly

Associated data


Grant support

The current study was sponsored by the National 12th Five Major Special Projects Funding Program (No. 2012ZX10001009) from the Ministry of Science and Technology of the People's Republic of China. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.