Expression and self-assembly of Grimsby virus: antigenic distinction from Norwalk and Mexico viruses

Clin Diagn Lab Immunol. 1999 Jan;6(1):142-5. doi: 10.1128/CDLI.6.1.142-145.1999.

Abstract

A cDNA obtained from Grimsby virus (GRV), a Norwalk-like virus, purified from a stool sample of a symptomatic adult associated with a gastroenteritis outbreak in the United Kingdom, was used to obtain the complete nucleotide sequence of the second open reading frame (ORF2). The ORF2 sequence of GRV predicts a capsid of 539 amino acids (aa) which exhibits aa identities of 96% to Lordsdale virus, 67% to Mexico virus (MXV), and 43% to Norwalk virus (NV). The GRV capsid protein was expressed in insects cells by using a recombinant baculovirus, and the resulting virus-like particles (VLPs) possessed a protein with an apparent molecular weight of 58,000. Hyperimmune antisera raised against purified GRV, MXV, and NV VLPs were tested in an indirect enzyme-linked immunosorbent assay (ELISA) against GRV, NV, and MXV VLPs, revealing that GRV is antigenically distinct from both NV and MXV. The antigenic specificity of the GRV-hyperimmune antiserum was confirmed in an antigen capture ELISA using GRV-, NV-, or MXV-containing fecal specimens. The expression of the GRV capsid protein has, for the first time, allowed the antigenic comparison of three distinct recombinant Norwalk-like viruses.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Amino Acid Sequence
  • Antibodies, Viral
  • Antigens, Viral / chemistry
  • Antigens, Viral / genetics*
  • Caliciviridae / genetics*
  • Caliciviridae / growth & development
  • Caliciviridae / immunology*
  • Caliciviridae Infections / virology
  • Capsid / chemistry
  • Capsid / genetics
  • Capsid / immunology
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • DNA, Viral / genetics
  • DNA, Viral / isolation & purification
  • Gastroenteritis / virology
  • Gene Expression
  • Genes, Viral
  • Humans
  • Microscopy, Electron
  • Molecular Sequence Data
  • Molecular Weight
  • Norwalk virus / genetics*
  • Norwalk virus / growth & development
  • Norwalk virus / immunology*
  • Open Reading Frames
  • Sequence Homology, Amino Acid
  • Species Specificity

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • DNA, Complementary
  • DNA, Viral

Associated data

  • GENBANK/AJ004864