Development of a reverse transcription polymerase chain reaction procedurefor the detection of marine caliciviruses with potential application for nucleotide sequencing

A reverse transcription polymerase chain reaction (RT-PCR) procedure is des cribed for the detection of marine caliciviruses including vesicular exanth ema of swine virus (VESV), San Miguel sea lion virus (SMSV), bovine Tillamo ok virus (BCV Bos-1) and caliciviruses (CV) isolated from dolphin (Cetacean CV), gorilla (Primate CV) and rattlesnake (Reptile CV) using primers (1F a nd 1R) designed from the capsid-coding region of the viral genome. These pr imers were compared with those described by Neill, J.D, and Seal, B.S., 199 5: Development of PCR primers for specific amplification of two distinct re gions of the genomes of San Miguel sea lion and vesicular exanthema of swin e viruses, Mel. Cell. Probes 9, 33-38 (Hel1/Hel2), which had been designed from the 2C-like region of the calicivirus genome. Both sets proved to be e xtremely useful diagnostic tools for all of the known marine calicivirus se rotypes with the exception of three: SMSV-8 and -12 and mink CV suggesting that these three caliciviruses may belong to a different group. Neither of the two primer sets reacted with strains of the vesicular disease viruses o f foot-and-mouth disease (FMD), swine vesicular disease (SVD) or vesicular stomatitis (VS) nor with two feline caliciviruses (FCV). The 1F/1R primer s et has the advantage over the Hel1/Hel2 set in that it generates a larger P CR product for nucleotide sequence investigations and so provides greater o pportunity for identifying molecular differences between the viruses. (C) 1 999 Elsevier Science B.V. All rights reserved.