摘要
为探究贵州省猪流行性腹泻病毒(PEDV)流行毒株的遗传进化情况,对2017—2022年贵州省不同地区13份PEDV阳性腹泻样本进行ORF3全基因扩增、克隆、测序,使用DNAstar、MEGA 7.0软件,对所得序列核苷酸和氨基酸同源性以及突变和遗传进化情况进行分析。结果显示:13份PEDV阳性样品的ORF3基因序列全长均为675 bp,编码224个氨基酸;13条克隆序列的核苷酸和氨基酸同源性分别为95.1%~100%和96.4%~100%;13条克隆序列与经典毒株CV777株ORF3基因的核苷酸和氨基酸同源性分别为95.9%~97.3%和94.7%~96.4%,同源性较低;与经典毒株CV777株相比,13条克隆序列在G160A、C243T、C274T、G301A、C450T、A497G存在相同的核苷酸突变,存在4个相同的氨基酸位点突变(V21A、V54I、A101T、N166S);13条克隆序列所在毒株在遗传进化上全部属于GⅡ型,其中GZ220514、GZ342、GZ224、GZ219、GZ423隶属于GⅡb亚型,GZ210112、GZ210129、GZ210220、GZ210714、GZ407、GZ210525、GZ330、GZ370属于GⅡa亚型(表明13份PEDV阳性样品所含毒株均为变异毒株);同一基因型的PEDV毒株ORF3基因相对较保守,但不同基因型差异较大;对所有参比株ORF3基因推导的氨基酸序列与CV777株进行比对,分析认为可以将25S、70V/F、80F、107F/V/L氨基酸突变模式作为GⅡa亚型的分子标记。本试验为分析贵州省PEDV流行以及遗传变异情况提供了参考,并丰富了贵州省PEDV ORF3基因序列。
In order to investigate the genetic evolution of the prevailing strains of porcine epidemic diarrhea virus(PEDV)in Guizhou Province,13 PEDV-positive diarrhea samples were collected from different regions across the province from 2017 to 2022,and the ORF3 genes were amplified,cloned and sequenced,then the nucleotide and amino acid homology as well as mutation and genetic evolution were analyzed using DNAstar and MEGA 7.0 software.The results showed that,ORF3 gene sequences of 13 positive samples were all 675 bp in length,encoding 224 amino acids.The nucleotide and amino acid homology of 13 cloned sequences were 95.1%~100%and 96.4%~100%,respectively,those of the sequences and the classical strain,CV777,were 95.9%~97.3%and 94.7%~96.4%,respectively,with low homology.Compared with the classical strain of CV777,same nucleotide mutations of 13 cloned sequences were observed at G160A,C243T,C274T,G301A,C450T and A497G,and 4 same amino acid mutations(V21A、V54I、A101T、N166S)were available.All the strains which 13 cloned sequences originating from belonged to type GⅡin genetic evolution,in which,GZ220514,GZ342,GZ224,GZ219 and GZ423 fell into subtype GⅡb,while GZ210112,GZ210129,GZ210220,GZ210714,GZ407,GZ210525,GZ330 and GZ370 were classified as subtype GⅡa,indicating that all the strains contained by the 13 positive samples were variants.The ORF3 genes of PEDV strains were relatively conservative in the same genotype,but quite different depending on different genotypes.It was analyzed and determined that,based on the comparison of all reference strains and CV777,the amino acid mutation patterns of 25S,70V/F,80F and 107F/V/L could be used as molecular markers of subtype GⅡa.Future analysis on PEDV prevalence and genetic variation in Guizhou Province were supported by the reference provided in the experiment that enriched the ORF3 sequence of PEDV strains registered in the province.
作者
万娟
梁海英
曾智勇
汤德元
王彬
张婧旭
柳佳佳
边孟婷
黄书
潘向英
田红利
Wan Juan;Liang Haiying;Zeng Zhiyong;Tang Deyuan;Wang Bin;Zhang Jingxu;Liu Jiajia;Bian Mengting;Huang Shu;Pan Xiangying;Tian Hongli(College of Animal Science,Guizhou University,Guiyang,Guizhou 550025,China)
出处
《中国动物检疫》
CAS
2023年第4期19-24,92,共7页
China Animal Health Inspection
基金
贵州省科技支撑计划项目(黔科合支撑[2021]一般162项目)。
