摘要
为调查2023年黑龙江地区某鹅场中鹅星状病毒(goose astrovirus,GoAstV)的感染及遗传进化情况,采集发病鹅的内脏组织样品进行RT-PCR检测,检测阳性样本通过鹅胚接种进行病毒分离,并利用RT-PCR以及电镜进行鉴定,利用二代测序技术进行分离毒株的全基因组测序,运用MEGA6软件对ORF2基因进行遗传进化分析。结果显示,在15份样品中,GoAstV的阳性率为6.67%(1/15)。阳性样本处理后接种鹅胚进行病毒分离,盲传至第5代时,尿囊膜增厚,出现尿酸盐沉积,鹅胚在120 h内死亡率达到83.33%(20/24),死亡鹅胚尿囊液进行GoAstV RT-PCR检测,结果为阳性。电镜观察结果显示,可观察到30 nm左右的病毒粒子。提取第5代病毒尿囊液RNA进行全基因组测序,并对该毒株ORF2基因进行同源性分析和系统发育进化树分析,结果显示,鉴定的毒株与其他GoAstV的ORF2基因核苷酸同源性为54.7%到96.6%,与GoAstV LYG2株的核苷酸同源性最高,为96.6%,而其与GoAstV FLX株差异较大,为54.7%。系统发育进化树结果显示,试验鉴定的毒株属于GoAstV G2型,与火鸡星状病毒遗传关系较近。研究对黑龙江地区某鹅场GoAstV进行分离鉴定并对ORF2基因进行序列分析,为黑龙江省GoAstV遗传进化及其分子流行病学调查研究提供了基础数据。
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.
作者
徐烨
朱庆贺
高旭
孙东波
Xu Ye;Zhu Qinghe;Gao Xu;Sun Dongbo(College of Agronomy,Yanbian University,Yanji 133000;College of Animal Science and Technology,Heilongjiang Bayi Agricultural University)
出处
《黑龙江八一农垦大学学报》
2024年第2期50-56,106,共8页
journal of heilongjiang bayi agricultural university
基金
黑龙江省“双一流”特色学科建设项目(2042080001)。
