Isolation,Identification and ORF2 Gene Sequence Analysis of Goose Astrovirus from a Goose Farm in Heilongjiang Province in 2023
In order to investigate the infection and genetic evolution of goose astrovirus(GoAstV)in a goose farm in Heilongjiang province in 2023,internal tissue samples of infected geese were collected for RT-PCR detection.Positive samples were isolated by goose embryo inoculation,identified by RT-PCR and electron microscopy.The whole genome of the isolated strain was sequenced by the second generation sequencing technology,and the genetic evolution of ORF2 gene was analyzed by MEGA6 software.The results showed that the positive rate of GoAstV was 6.67%(1/15)in 15 samples.After the positive samples were treated,the goose embryo was inoculated for virus isolation,the allantoic membrane thicken and urate deposition occurred at the 5 th generation of blind transmission.The mortality rate of the goose embryo reached 83.33%(20/24)within 120 h.The allantoic liquid of the dead goose embryo was detected by GoAstV RT-PCR and the result was positive.Electron microscopy showed that virions of about 30 nm could be observed.Allantoic fluid RNA of the 5 th generation virus was extracted for whole genome sequencing,and homology analysis and phylogenetic evolutionary tree analysis of ORF2 gene of this strain were conducted.The results showed that the nucleotide homology of the identified strain with ORF2 gene of other GoAstV was 54.7%to 96.6%,and the nucleotide homology with LYG2 strain of GoAstV was the highest.It was significantly different from GoAstV FLX(54.7%)and 96.6%.Phylogenetic tree results showed that the strain identified in this study belonged to GoAstV G2 type,it was closely related to Turkey astrovirus.In this study,the test isolated and identified GoAstV from a goose farm in Heilongjiang province and analyzed the ORF2 gene sequence,which provided basic data for the genetic evolution and molecular epidemiology of GoAstV in Heilongjiang province.
国家科技期刊平台
NSTL
万方数据
