摘要
为建立高效准确的猪繁殖与呼吸综合征(PRRS)快速确诊的检测方法,基于RT-PCR的焦磷酸测序检测方法,采用PSQ Assay Design SW软件分析PRRSV N蛋白基因的保守序列,设计扩增引物和测序引物,经过RT-PCR扩增和焦磷酸测序后,通过直观的序列结果显示,可直接进行PRRSV的确证,所建立的焦磷酸测序检测方法特异性好,且提高了检出率和准确性,避免了假阳性的出现,为猪繁殖与呼吸综合征的快速确诊提供了技术支撑,建立了PRRSV N蛋白基因的焦磷酸测序方法。
In order to establish an efficient and accurate detection method for the rapid diagnosis of porcine reproductive and respiratory syndrome(PRRS),based on the pyrosequencing detection method based on RT-PCR,PSQ assay design SW software was used to analyze the conserved sequence of PRRSV N protein gene,and amplification primers and sequencing primers were designed.After RT-PCR amplification and pyrosequencing,the intuitive sequence results showed that,The established pyroseque ncing method has good specificity,improves the detection rate and accuracy,avoids the occurrence of false positive,and provides technical support for the rapid diagnosis of porcine reproductive and respiratory syndrome.A pyrosequencing method of PRRSV N protein gene was established.
作者
曹丙蕾
张群
刘敏
王群
尹伟力
Cao Binlei;Zhang Qun;Liu Min;Wang Qun;Yin Weili(Ji'nan Customs Technology Center,Jinan 250014,Shandong,China;Qingdao Customs Technology Center,Qingdao 264001,Shandong,China)
出处
《农业技术与装备》
2021年第12期142-143,145,共3页
Agricultural Technology & Equipment
基金
畜禽疫病液相芯片高通量检测技术及东部沿海口岸监测应用示范(2016YFD050110401)
原国家质检总局科研项目(2016IK193)
原山东出入境检验检疫科研项目(SK201401)(SK201713)
济南海关科研项目(2019JK004)。
关键词
猪繁殖与呼吸综合征病毒
焦磷酸测序
PCR
鉴定
porcine reproductive and respiration syndrome virus
pyrosequencing assay
PCR
identification
