Calicivirus recombinant expressing fusion protein reacting to multiple calicivirus typing sera

Abstract

The Caliciviridae contains many viruses which are pathogenic to humans, marine mammals, domestic animals, and numerous species of wildlife. Currently there is no single assay to detect antigenic response to the multiple serotypes of calicivirus. The development of calicivirus specific synthetic peptides having highly conserved epitopes common to many serotypes would facilitate the development of a simple and rapid serologic assay for calicivirus antibodies irrespective of the serotype. Calicivirus cDNA recombinants which express fusion proteins that react with multiple calicivirus typing sera may be useful in the development of a serologic assay for antibodies to caliciviruses. For this purpose RNA was isolated from cell culture infected with San Miguel sea lion virus type 5 (SMSV-5) and used to construct a cDNA library, named SMSV-5 lambda. Immunoassay techniques were used to screen the SMSV-5 lambda library and a second cDNA library, named SMSV-5RT, also constructed from SMSV-5. One recombinant named 8-SN was identified which produced a fusion protein that reacted positively with a pool of four polyclonal calicivirus typing sera (SMSV-5, SMSV-13, SMSV-15, and SMSV-17). This construct was amplified, induced, and a fusion protein identified which reacted positively in four western blot assays using individual polyclonal typing sera to the caliciviruses SMSV-13, SMSV-15, SMSV-16, and SMSV-17.