To isolate porcine rotavirus type A,16 fecal samples and small intestine contents of PRoV positive piglets collected from different regions were treated and inoculated with MA-104 cells.After passage,fluorescence quantitative PCR(qPCR),indirect immunofluorescence assay(IFA),electron microscopy and sequence analysis were used for identification.The results showed that the Ct value of F1-F3 generation virus decreased gradually with the passage increase,the specific fluorescence could be observed by IFA,and the diameter of 70nm virus particles without envelope was observed by electron microscope.Subsequently,seven strains of PRoV were isolated,of which 4 strains were G9,2 strains were G3,and 1 strain was G4.Homology analysis and genetic evolution analysis based on VP7 gene were performed on 7 isolates and other 9 positive samples.The results showed that 16 samples contained 3 G genotypes,namely G9(7/16),G4(6/16)and G3(3/16).Meanwhile,G9 strain FJ-2021 and G4 strain JS01-2021 were close to the prevalent strains in recent years.The geographical distribution of G genotypes in 16 samples was further analyzed and showed that East China had the prevalence of G9,G4 and G3 genotypes,Sichuan had prevalence of G3 and G9 genotypes,and Guangdong had prevalence of G9 genotypes,which were consistent with the prevalence of G genotypes previously reported in East China,Sichuan,and Guangxi,respectively.In addition,the G9 genotype is prevalent in Yunnan,Liaoning and Hunan,and the G4 genotype is prevalent in Guangxi,Guangdong,Shaanxi,Heilongjiang and Henan.This study,through the isolation and identification of PRoV positive clinical samples from 12 provinces and cities,has enriched the epidemic data of PRoV in China.It provided epidemiological insights for targeted prevention and control of PRoV across different regions,and lays a foundation for further research on the pathogenesis and vaccine development of different G geno types of PRoV.
NETL
NSTL
万方数据
