A capsid gene-based real-time reverse transcription polymerase chain reaction assay for the detection of marine vesiviruses in the Caliciviridae.
A real-time reverse transcription polymerase chain reaction (rtRT-PCR) assay was developed for the identification of marine vesiviruses. The primers were designed to target a 176-nucleotide fragment within a highly conserved region of the San Miguel sea lion viruses (SMSVs) capsid gene. The assay detected viral RNA from nine marine vesivirus serotypes described previously, including two serotypes (SMSV-8 and -12) not identified with presently available molecular assays, a highly related bovine vesivirus strain (Bos-1), a mink vesivirus strain (MCV), and two novel genotypes isolated recently from Steller sea lions (SSL V810 and V1415). The real-time assay did not amplify sequences from the corresponding genomic regions of feline calicivirus (also in the genus Vesivirus) and representative members of the genus Norovirus. The rtRT-PCR assay described below may prove useful as a diagnostic tool for the detection of currently circulating, emerging and previously described marine vesiviruses in clinical samples, especially when large numbers are screened in surveillance studies of these restricted viruses.
McClenahan
Shasta D
SD
Department of Infectious Diseases and Pathology, College of Veterinary Medicine, University of Florida, 2015 SW 16th Avenue, Gainesville, FL 32610, USA.
Bok
Karin
K
Neill
John D
JD
Smith
Alvin W
AW
Rhodes
Crystal R
CR
Sosnovtsev
Stanislav V
SV
Green
Kim Y
KY
Romero
Carlos H
CH
D000818:Animals; D001483:Base Sequence; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D036022:Capsid Proteins / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D029342:Vesivirus / Q000235:genetics / Q000302:isolation & purification*; D014871:Water Microbiology
D036022:Capsid Proteins; D017931:DNA Primers; D012367:RNA, Viral