分离新型鸭源微RNA病毒进行全基因组测序并进行遗传进化分析.对本实验室2021年不同来源的种鸭和肉鸭病料进行PCR检测,初步确定存在一种未知分类的新型微RNA病毒感染.取病死鸭病料组织处理后接种SPF鸡胚分离病毒,设计引物对分离到的病毒进行PCR检测,通过重叠PCR方法进行全基因组扩增测序.将分离病毒各蛋白氨基酸序列两两比对,同时选取GenBank数据库中微RNA病毒代表毒株序列绘制系统进化树,并对主要蛋白P1、2C、3D序列比对分析.结果显示:共分离到三株微RNA病毒,分别命名为21101株、21016株和21075株(GenBank登录号:OQ927377～OQ927379).基因组长度分别为7 445、7 445和7 447 bp,均包含一个编码2 141个氨基酸的开放阅读框(ORF),可划分为P1、P2、P3三个部分,符合微RNA病毒序列特征.基于全基因组序列遗传进化分析发现,三株分离病毒与本实验室前期分离的Duck/FC22/China/2017(GenBank登录号:MN102111)毒株及上海兽医研究所分离的Duck/AH15/CHN/2015(GenBank登录号:MT681985)位于同一分支,与鸭甲型肝炎病毒(Duck hepatitis A virus,DHAV)遗传距离最近.分离的三株鸭源微RNA病毒进行全基因组测序及遗传进化分析发现,与目前已知的两株微RNA毒株为同一类新型鸭源微RNA病毒.
Isolation and Identification of Novel Picornavirus from Ducks and Whole Genome Sequence Analysis
The aim of this study was to isolate novel duck-derived picornavirus for whole genome sequencing and genetic in order to perform whole genome sequencing and genetic evolutionary study.In 2021,we identified the presence of an unknown class of novel picornavirus infection in the diseased materials of breeding ducks and meat ducks submitted for inspection.By inoculating the diseased material into specific pathogen-free(SPF)chicken embryos,we isolated the virus.And the isolated viruses were further detected by PCR with primers designed from sequences available and sequenced the whole genome by overlapping PCR method.The amino acid sequences of each protein of the isolated virus were compared.At the same time,the sequences of the representative strains of small RNA viruses in the GenBank database were selected to draw the phylogenetic tree,and the sequences of the main proteins P1,2C and 3D were analysed in comparison with each other.Three small RNA virus strains were isolated and named 21101,21016 and 21075(GenBank ID:OQ927377-OQ927379),with genome lengths of 7 445,7 445,and 7 447 bp.These sequence encompasses an open reading frame(ORF)that encodes a total of 2 141 amino acids.The ORF can be divided into three distinct parts:P1,P2 and P3.These protein divisions align with the characteristic features observed in picornaviruses.The genetic evolutionary analysis,conducted through the utilization of whole genome sequences,demonstrated that the three isolates were positioned within the same branch as the Duck/FC22/China/2017 strain(GenBank ID:MN102111)previously separate inour laboratory and Duck/AH15/CHN/2015 strain(GenBank ID:MT681985)at the Shanghai Veterinary Research Institute.Furthermore,these isolates exhibited the highest degree of genetic similarity to the Duck hepatitis A virus(DHAV).Three strains of duck-derived picornavirus were isolated,and whole genome sequencing and genetic evolution analysis revealed that the three isolated viruses belong to the same class of the two currently known novel duck-derived picornavirus.
novel picornavirusisolation and identificationgenome sequencegenetic evolutionary analysis
国家科技期刊平台
NSTL
万方数据
