Identification and genomic characteristics of a novel caprine enterovirus stain
In this study,we identified the mixed infection of peste des petits ruminants virus(PPRV)and CEV from the lymphoid tissue of a dead sheep by metagenome sequencing technology.To isolate CEV,the total viral RNA from the mesenteric lymph node tissue was used to transfect into BHK21 cells.After continuous passage,the virus was isolated and identified by cytopathogenic observation,sequencing,TCID50,one-step growth curve of virus,and electron microscope observation.The genetic characteristics of the isolated virus were compared with the reference strains.The results indicated that BHK21 cells transfected with total viral RNA showed obvious CPE in the second passage and sequencing showed that the culture only contained CEV,and its total genome length was 7 446 nt,which possess 5'UTR length of 821 nt,3'UTR of 109 nt and the ORF of 6 516 nt.The isolated virus was named JSNT/1/2022 strain and divided into enterovirus G(EV-G),compared with the reference strains.The virus particle was measured 2030 nm in diameter,and the TCID50 value of the 6 th generation virus was 104,2/0.1 mL at 8 h.In this study,a strain of CEV was firstly isolated by transfection of tissue material RNA into cells,indicating that the nucleic acid of CEV has infectious,which provides a more reliable method for the isolation and identification of CEV.The isolated virus will lay the etiological foundation for the basic and applied research on CEV.
caprine enterovirusisolation and identificationRNA infectivitymacrogenomic sequencing
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