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Characterization of porcine sapelovirus isolated from Japanese swine with PLC / PRF /5 cells
Author(s) -
Bai H.,
Liu J.,
Fang L.,
Kataoka M.,
Takeda N.,
Wakita T.,
Li T.C.
Publication year - 2018
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/tbed.12796
Subject(s) - open reading frame , untranslated region , vero cell , microbiology and biotechnology , biology , virology , cell culture , cytopathic effect , peptide sequence , nucleic acid sequence , virus , gene , messenger rna , genetics
Summary Porcine sapelovirus ( PSV ) is a causative agent of neurological disorders, fertility disorders and dermal lesions of swine. In this study, we isolated two PSV strains, Jpsv477 and Jpsv1315, from swine faecal specimens using a PLC / PRF /5 cell culture system. The PSV infection of PLC / PRF /5 cells induced a cytopathic effect ( CPE ). Two types of virus particles with identical diameter (~35 nm) but different densities (1.300 and 1.285 g/cm 3 ) were observed in the cell culture supernatants. Analysis of the entire genome sequence of Jpsv477 and Jpsv1315 revealed that both strains possess 7,558 nucleotides and the poly (A) tail and have a typical PSV genome organization consisting of a 5′ terminal untranslated region (5′ UTR ), a large open reading frame ( ORF ), and a 3′ terminal untranslated region (3′ UTR ). The ORF encodes a single polyprotein that is subsequently processed into a leader protein (L), four structural proteins ( VP 1, VP 2, VP 3 and VP 4) and seven functional proteins (2A, 2B, 2C, 3A, 3B, 3C and 3D). The structural proteins VP 1, VP 2, VP 3 and VP 4 have molecular masses of ~35, ~26, ~25 and ~6 kDa. The N‐terminal amino acid sequence analysis of VP 1, VP 2, VP 3 and VP 4 confirmed that the cleavage sites between VP 4 and VP 2, VP 2 and VP 3, and VP 3 and VP 1 are K/A, Q/G and Q/G, respectively. We further confirmed that HepG2/C3A, Vero E6 and primary green monkey kidney cells ( PGMKC ) were also susceptible to PSV infection. The stability assay demonstrated that PSV was inactivated by heating at 60°C for 10 min or 65°C for 5 min. The virus also lost infectivity by incubation with 62.5 ppm of NaClO for 30 min.