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Clinical and Diagnostic Laboratory Immunology, January 1999, p. 142-145, Vol. 6, No. 1
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Expression and Self-Assembly of Grimsby Virus: Antigenic Distinction from Norwalk and Mexico Viruses

Antony D. Hale,1,2 Sue E. Crawford,1 Max Ciarlet,1 Jonathan Green,2 Christopher Gallimore,2 David W. G. Brown,2 Xi Jiang,3 and Mary K. Estes1,*

Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 770301; Enteric and Respiratory Virus Laboratory, Central Public Health Laboratory, London NW9 5HT, United Kingdom2; and Center for Pediatric Research, Children's Hospital of The King's Daughters, Eastern Virginia Medical School, Norfolk, Virginia 235103

Received 10 August 1998/Returned for modification 2 October 1998/Accepted 6 November 1998

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.

* Corresponding author. Mailing address: Division of Molecular Virology, Baylor College of Medicine, Mailstop 739E, One Baylor Plaza, Houston, TX 77030. Phone: (713) 798-3585. Fax: (713) 798-3586. E-mail: mestes{at}bcm.tmc.edu.

Clinical and Diagnostic Laboratory Immunology, January 1999, p. 142-145, Vol. 6, No. 1
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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