Characterization of San Miguel sea lion virus populations using pyrosequencing-based methods.
San Miguel Sea Lion Virus (SMSV) is a small RNA virus in the genus Vesivirus with an unusually broad host range. Three populations of SMSV were examined by PCR amplification of the capsid precursor and putative helicase genes, followed by pyrosequencing. The populations were nasal swabs from two SMSV infected California sea lions (Zalophus californianus) from two different years, and a virus isolate from the earlier swab that was passaged in cell culture five times. In the capsid precursor, extensive deletions were prevalent in the passaged virus but uncommon in the clinical samples. A greater prevalence of point mutations was seen in the capsid precursor gene than in the putative helicase gene. In culture, the minority sequence in the capsid precursor at nucleotide position 5826 rapidly shifted after five passages to become the majority sequence. Levels of diversity at individual sites showed much more similarity between the two clinical samples than between the earlier clinical sample and the passaged culture from the same sample. SMSV appears to behave as a quasispecies. Assessment of original patient samples is preferable for understanding clinical SMSV populations.
Wellehan
James F X
JF
Marine Animal Disease Laboratory, College of Veterinary Medicine, University of Florida, 2015 SW 16th Ave, Gainesville, FL 32610, USA. wellehanj@vetmed.ufl.edu <wellehanj@vetmed.ufl.edu>
Yu
Fahong
F
Venn-Watson
Stephanie K
SK
Jensen
Eric D
ED
Smith
Cynthia R
CR
Farmerie
William G
WG
Nollens
Hendrik H
HH
D000818:Animals; D017250:Caliciviridae Infections / Q000662:veterinary / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics; D004252:DNA Mutational Analysis; D019143:Evolution, Molecular; D016679:Genome, Viral / Q000235:genetics*; D009296:Nasal Cavity / Q000821:virology; D046070:Sea Lions / Q000821:virology*; D029342:Vesivirus / Q000235:genetics* / Q000302:isolation & purification; D014764:Viral Proteins / Q000235:genetics*
D036022:Capsid Proteins; D014764:Viral Proteins