猪流行性腹泻病病毒RT-PCR分子诊断及部分S基因同源性分析
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摘要
2018年3月,河南焦作市某猪场发生疑似猪流行性腹泻(PED)感染,而该猪场进行过PED疫苗免疫,为确诊病原,采集患病仔猪肠管内容物,利用RT-PCR法,扩增猪流行性腹泻病毒(PEDV)部分S基因片段,并将产物送基因公司测序。所获测序结果,运用DNAStar和MEGA5软件,与GeneBank下载的不同毒株S基因片段进行同源性分析。结果显示:RT-PCR扩增产物与预期大小一致,为969bp;该毒株部分S基因与经典毒株CV777同源性为94.8%,亲缘关系较远;与近年来流行毒株同源性为98.1%～99.1%,亲缘关系较近。PEDV变异毒株的出现可能是导致免疫失败的重要原因。
In March 2018,a suspected Porcine Epidemic Diarrhea(PED) infection occurred in a pig farm in Jiaozuo,Henan Province.After our investigation,the pigs have been immunized with PED vaccine.To diagnose the cause of the infection,the intestinal contents of infected piglets was collected and RT-PCR was used to amplify the S gene segment of porcine epidemic diarrhea virus(PEDV), then the amplified S gene segment was sent to the gene company for sequencing.The sequencing result was obtained,The DNAStar and MEGA5 software were used to perform homology analysis with the S gene fragment of different strains downloaded from GeneBank.The results showed that the RT-PCR amplification product had the expected size of 969 bp;The homology of the S gene to the classical strain CV777 was 94.8% and the genetic relationship was far away;It has a homology of 98.1% to 99.1% with the prevalent strains in recent years and has a close genetic relationship.The emergence of PEDV variant strains may be the root cause of immune failure.
In March 2018,a suspected Porcine Epidemic Diarrhea(PED) infection occurred in a pig farm in Jiaozuo,Henan Province.After our investigation,the pigs have been immunized with PED vaccine.To diagnose the cause of the infection,the intestinal contents of infected piglets was collected and RT-PCR was used to amplify the S gene segment of porcine epidemic diarrhea virus(PEDV), then the amplified S gene segment was sent to the gene company for sequencing.The sequencing result was obtained,The DNAStar and MEGA5 software were used to perform homology analysis with the S gene fragment of different strains downloaded from GeneBank.The results showed that the RT-PCR amplification product had the expected size of 969 bp;The homology of the S gene to the classical strain CV777 was 94.8% and the genetic relationship was far away;It has a homology of 98.1% to 99.1% with the prevalent strains in recent years and has a close genetic relationship.The emergence of PEDV variant strains may be the root cause of immune failure.
引文
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[3]王贵华,于萍萍,满坤,等.五株猪流行性腹泻病毒的分离鉴定及序列分析[J].中国兽药杂志,2016,50(07):13-19.
[4]陈如敬,吴学敏,车勇良,等.猪流行性腹泻病毒FJ-11A株的分离与ORF3基因序列分析[J].福建农业学报,2011,26(06):947-951.
[5]杜晓莉,王一成,吴润,等.2010—2013年浙江省猪流行性腹泻病毒临床检测及PEDV-S基因型分析[J].浙江农业学报,2014,26(03):581-587.
[6]朱迪国,宋建德,袁丽萍,等.2013—2014年全球猪流行性腹泻重大疫情分析[J].中国动物检疫,2014,31(10):42-46.
[7]刘政龙,王金良,沈志强.猪流行性腹泻病毒研究进展[J].中国畜牧兽医,2015,42(09):2506-2511.
[8]王二鑫,张春杰,廖成水,等.2016年河南省猪流行性腹泻病毒S、M基因遗传变异分析[J].中国预防兽医学报,2018,40(03):256-259.
[9]董建国,王瑞,曲哲会,等.2014—2015年豫南地区猪流行性腹泻病毒S基因变异分析[J].畜牧兽医学报,2018,49(04):859-864.
[10]尹宝英,吴旭锦,熊忙利.猪流行性腹泻诊断方法研究进展[J].动物医学进展,2013,34(12):156-159.
[11]李舜,王爱富,李桂珍,等.2015年～2016年广东省猪流行性腹泻病毒部分S1基因的遗传进化分析[J].中国预防兽医学报,2018,40(03):253-255.
[12]吴旺霞,王一成,吴润,等.2011—2016年浙江省猪流行性腹泻病毒S基因与ORF3基因遗传进化分析[J].浙江农业学报,2017,29(03):380-388.
[13]袁定胜.浅谈猪流行性腹泻的诊断与防治措施[J].畜禽业,2018,29(01):6-7.
[14]徐国栋,李峰,张广峰.国内猪流行性腹泻防治概况[J].畜牧与兽医,2011,43(12):88-93.