贵州省猪流行性腹泻病毒ORF3、M基因克隆及序列分析

Cloning and Sequence Analysis of ORF3 and M Gene of Porcine Epidemic Diarrhea Virus in Guizhou Province

为了解贵州省猪流行性腹泻病毒（porcine epidemic diarrhea virus,PEDV）毒株ORF3及M基因的遗传变异情况,试验于2014年4月-2015年3月从贵州省5个地区采集105份腹泻仔猪的粪便,应用RT-PCR方法进行PEDV检测,从中选择8份PEDV阳性样本,扩增其ORF3及M基因,测序并进行序列比对分析。结果显示,从采集的105份粪便样本中可检出75份PEDV阳性样本,阳性率为71.43%;8株PEDV贵州株ORF3及M基因序列均无碱基缺失或插入;ORF3基因核苷酸及推导的氨基酸同源性在95.1%-100.0%与95.1%-99.6%之间,M基因核苷酸及推导的氨基酸同源性在98.4%-100.0%与98.7%-100.0%之间;氨基酸系统进化树分析结果显示,2014-2015年贵州流行株与近年来中国毒株、韩国毒株及泰国毒株亲缘关系较近,与疫苗株Attenuated DR13及CV777株亲缘关系较远。提示目前贵州省仔猪腹泻病原主要是PEDV,且为PEDV强毒株。

To study the genetic variations of porcine epidemic diarrhea virus（PEDV）ORF3and M gene in Guizhou province,we used RT-PCR method to detect PEDV in the dung what collected from diarheal porket in five regions of Guizhou province between April 2014 to March 2015,then selected eight positive samples,cloned and sequenced their ORF3 and M gene.The results showed that 75 samples were positive for PEDV,and the positive rate was 71.43%.The result of sequencing showed that ORF3 and Mgene were intact;ORF3gene shared from 95.1%to 100.0% nucleotide identity and 95.1% to 99.6% amino acid identity,and M gene shared from 98.4% to100.0% nucleotide identity and 98.7%to 100.0% amino acid identity with eight PEDV Guizhoustrains.Phylogenetic analysis revealed that Guizhou strains seem to be closely related to Chinese strains,Korean strains and Thai strains,and there were genetically different from the vaccine strains attenuated DR13 and CV777.The results suggested that in rencent years the mainly etiology of orket diarrhea was velogenic PEDV.