RESUMO
The human hepatocarcinoma cell line PLC/PRF/5 is susceptible to hepatitis E virus (HEV) infection and is used for HEV isolation. It is difficult to use the cell line for this purpose directly from fecal specimens of swine or wild boar contaminated with porcine sapelovirus (PSV) because PSV infection results in rapid and extensive cytopathic effects in PLC/PRF/5 cells, interrupting the growth of HEV. Herein, we used a PSV infection-resistant cell line, N1380, derived from PLC/PRF/5 cells, and successfully isolated a HEV-4b strain from a PSV-positive swine fecal specimen. Our results indicated that N1380 cells are a useful tool for the isolation of HEV from swine or wild boar fecal specimens, even when the cells are co-infected with PSV.
Assuntos
Fezes/virologia , Vírus da Hepatite E/classificação , Vírus da Hepatite E/isolamento & purificação , Hepatite E/diagnóstico , Doenças dos Suínos , Animais , Linhagem Celular , Hepatite E/veterinária , Vírus da Hepatite E/genética , Picornaviridae/isolamento & purificação , Infecções por Picornaviridae/diagnóstico , SuínosRESUMO
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.
Assuntos
Fezes/virologia , Genoma Viral/genética , Macaca fascicularis/virologia , Picornaviridae/genética , Vírion/genética , Regiões 3' não Traduzidas/genética , Regiões 5' não Traduzidas/genética , Células A549 , Animais , Sequência de Bases/genética , Linhagem Celular Tumoral , Chlorocebus aethiops , Células HEK293 , Células Hep G2 , Humanos , Fases de Leitura Aberta/genética , Filogenia , Picornaviridae/classificação , Picornaviridae/isolamento & purificação , Análise de Sequência de DNA/métodos , Células Vero , Vírion/isolamento & purificaçãoRESUMO
By MAC-ELISA technique, Japanese encephalitis infection incidence was determined in HaTay as high in summer months of May, June, July in consisting with that of epidemic season in North VietNam. In Tay Nguyen (Central Highland), the incidence of Japanese encephalitis was low in pig’s population, dispersing in all the year round, similarly as the epidemic cases identified in 1998-2002 year period. By UCNKHC (?) Japanese high level of encephalitis antibody in population was determined in all round 12 months. No relationship of Japanese encephalitis infection in pig’s population and Japanese encephalitis cases was identified
