一株G4P[X]型猪A群轮状病毒的分离与鉴定

Isolation and Identification of One Strain of G4P[X]Porcine A Group Rotavirus

为了分离并鉴定一株A群猪轮状病毒(PoRV)。本文通过MA-104细胞分离培养、间接免疫荧光鉴定、RT-PCR鉴定以及全基因序列遗传进化对一份PoRV阳性粪便样本进行了分离与鉴定。经RT-PCR和间接免疫荧光证实为PoRV,命名为JC-3,基因型为G4-P[X]-I5-R1-C1-M1-A8-N1-T7-E1-H1,根据同源性比对分析发现,JC-3株VP1、VP6、VP7、NSP1、NSP2和NSP5基因与PoRV相似性最高,高达95.4%~99.7%,而VP2、VP3、NSP3、NSP4与人源A群轮状病毒(GAR)相似性最高,达94.6%~98.5%,表明该病毒基因组发生了重排。本研究成功分离一株猪源PoRV,基因型为G4P[X],为以后对该毒株的致病性研究奠定了基础。

To isolate and identify a group A porcine rotavirus(PoRV).A PoRV-positive stool sample was isolated and identified by MA-104 cell isolation and culture,indirect immunofluorescence identification,RT-PCR identification and sequence analysis.After screening the virus strain by ordinary PCR,PoRV was confirmed by RT-PCR and indirect immunofluorescence,named JC-3,and the genotype was G4-P[X]-I5-R1-C1-M1-A8-N1-T7-E1-H1.Homology analysis showed that the gene identities of VP1,VP6,VP7,NSP1,NSP2 and NSP5 of JC-3 strain were the highest with PoRV,up to 95.4%~99.7%,while VP2,VP3,NSP3 and NSP4 had the highest similarity with human group A rotavirus(GAR),up to 94.6%~98.5%,indicating that the virus may be a human-pig reassortant strain and may have undergone recombination phenomenon.However,no recombination phenomenon was found by recombination analysis.In this study,we successfully isolated a pig-derived PoRV with genotype G4P[X],which laidthe foundation for subsequent studies on its pathogenicity.