Abstract
We isolated a novel simian sapelovirus (SSV), Cam13, from fecal specimen of a cynomolgus monkey by using PLC/PRF/5 cells. The SSV infection of the cells induced an extensive cytopathic effect. Two types of virus particles with identical diameter (~32 nm) but different densities (1.348 g/cm3 and 1.295 g/cm3) were observed in the cell culture supernatants. The RNA genome of Cam13 possesses 8,155 nucleotides and a poly(A) tail, and it has a typical sapelovirus genome organization consisting of a 5’ terminal untranslated region, a large open reading frame (ORF), and a 3’ terminal untranslated region. The ORF encodes a single polyprotein that is subsequently processed into a leader protein (L), four structural proteins (VP1, VP2, VP3, and VP4) and seven functional proteins (2A, 2B, 2C, 3A, 3B, 3C, and 3D). We confirmed that 293 T, HepG2/C3A, Hep2C, Huh7 and primary cynomolgus monkey kidney cells were susceptible to SSV infection. In contrast, PK-15, Vero, Vero E6, RD-A, A549, and primary green monkey kidney cells were not susceptible to SSV infection. We established an ELISA for the detection of IgG antibodies against SSV by using the virus particles as the antigen. A total of 327 serum samples from cynomolgus monkeys and 61 serum samples from Japanese monkeys were examined, and the positive rates were 88.4% and 18%, respectively. These results demonstrated that SSV infection occurred frequently in the monkeys. Since Cam13 shared 76.54%–79.52% nucleotide sequence identities with other known SSVs, and constellated in a separate lineage in the phylogeny based on the entire genome sequence, we propose that Cam13 is a new genotype of the simian sapelovirus species.
Highlights
Many simian viruses (SV) were isolated from various primate tissues during the development of tissue culture methods as well as from specimens derived from primates used in biomedical research[1,2,3,4,5,6,7]
When we used the sterile-filtered 10% stool suspensions to inoculate PLC/PRF/5 cells, an unexpected cytopathic effect (CPE) was observed on day 7 post-inoculation (p.i.) in the cells that received the suspension from 1 of 10 monkeys imported from Cambodia (C13), no hepatitis E virus (HEV) RNA was detected in the supernatant
The CPE was clearly observed at day 2 p.i., and again no HEV RNA was detected in the supernatant, indicating that the CPE was unlikely to be caused by HEV infection
Summary
Many simian viruses (SV) were isolated from various primate tissues during the development of tissue culture methods as well as from specimens derived from primates used in biomedical research[1,2,3,4,5,6,7] Most of these viruses have been classified into the genus Enterovirus of the family Picornaviridae[8,9]. Based on the entire genome sequences, SV2 and PEV8 were classified into a new genus Sapelovirus[11], which currently consists of two species: Sapelovirus A (formerly known as porcine sapelovirus [PSV]), and Sapelovirus B (formerly named as simian sapelovirus [SSV])[12]. We proposed the introduction of a genotyping system to classify SSV isolates among Sepelovirus B
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