摘要
猪繁殖与呼吸综合征病毒(PRRSV)感染肺泡巨噬细胞,严重损害免疫系统,导致与其他病毒共感染现象,其中猪流感病毒(SIV)也是病原之一。为了快速检测和区分PRRSV基因型与SIV亚型,本研究针对H1、H3亚型SIV的HA基因和美洲型、欧洲型PRRSV的N、M基因的保守序列,分别设计了4对特异性扩增引物,在引物浓度、退火温度等反应条件的系统优化下,分别对其特异性和敏感性进行验证,建立了能够快速检测以上4种病原的多重PCR方法,并对155份临床样品进行了检测。结果显示,该检测方法特异性好、灵敏度高,对H1-SIV、H3-SIV、NA-PRRSV和EU-PRRSV的最低检出量均低至9.86×10^(0) copies/μL。以单一PCR方法对所有临床样品进行检测后发现,检测结果与多重PCR检测结果一致,证明这种多重PCR方法可以用于这4种病毒的快速检测与分型鉴定。
Porcine reproductive and respiratory syndrome virus(PRRSV)infects alveolar macrophages,seriously damaging the immune system,leading to co-infection with other viruses,among which swine influenza virus(SIV)is also one of the pathogens.In order to rapidly detect and distinguish PRRSV genotypes and SIV subtypes,four pairs of specific primers were designed according to the conserved sequences of HAgenes of H1 and H3 subtype SIV and Nand Mgenes of the American and European PRRSVs.The primers were optimized for their concentrations and annealing temperatures and the specificity and sensitivity of this multiplex PCR method were also demonstrated for rapidly detecting these four pathogens.The detection limit of H1-SIV,H3-SIV,NA-PRRSV and EU-PRRSV was 0 as low as 9.86×10^(0) copies/μL.A total of 155 clinical samples were then tested using this multiplex PCR method and single PCR methods.The results were consistent among these PCR methods,indicating that this multiplex PCR method could be used for rapid detection and type identification of these four viruses.
作者
徐毅
余良政
林霜
任同伟
王豪
曾昊
郭嘉宁
郭金凡
黄崇强
欧阳康
黄伟坚
韦祖樟
陈樱
XU Yi;YU Liangzheng;LIN Shuang;REN Tongwei;WANG Hao;ZENG Hao;GUO Jianing;GUO Jinfan;HUANG Chongqiang;OUYANG Kang;HUANG Weijian;WEI Zuzhang;CHEN Ying(Laboratory of Animal Infectious Diseases and Molecular Immunology,College of Animal Science and Technology,Guangxi University,Nanning 530004,China)
出处
《中国动物传染病学报》
CAS
北大核心
2024年第1期135-141,共7页
Chinese Journal of Animal Infectious Diseases
基金
广西创新驱动发展专项资金项目(桂科AA17204057-1)。
关键词
猪流感病毒
猪繁殖与呼吸综合征病毒
多重PCR
Swine influenza virus
Porcine reproductive and respiratory syndrome virus
multiplex PCR
