小反刍兽疫病毒特异性IgY抗体的制备及双抗夹心ELISA检测方法的建立
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摘要
本研究制备了针对小反刍兽疫PPRV全病毒特异性卵黄抗体,利用PPRV N蛋白特异性单克隆抗体和PPRV IgY为主要材料,建立了PPRV双夹心ELISA检测方法检测小反刍兽疫病毒。用该ELISA方法分别检测小反刍兽疫病毒、蓝舌病病毒、鹿流行性出血病病毒、水泡性口炎病毒、赤羽病病毒、口蹄疫病毒,结果表明该ELISA方法可以特异性检出PPRV而与其他病毒间无交叉。用该ELISA和RT-PCR同时检测162份临床样品,结果表明ELISA的特异性和敏感性分别为99.2%和93.7%,两种方法的符合率为98.1%。该方法的建立为小反刍兽疫病毒的初筛检测及小反刍兽疫流行病学调查提供了经济、快速、有效的方法,适用于设备条件不足的基层实验室。
引文
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[8]BATTEN C A,BANYARD A C,KING D P,et al.A real time RT-PCR assay for the specific detection of Peste des petits ruminants virus[J].Journal of Virological Methods,2011,171(2):401-404.
[9]ABERA T,THANGAVELU A,JOY CHANDRAN N D,et al.A SYBR Green I based real timeRT-PCR assay for specific detection and quantitation of Peste des petits ruminants virus[J].BMC Veterinary Research,2014,10:22.
[2]BANYARD A C,PARIDA S,BATTEN C,et al.Global distribution of peste des petits ruminants virus and prospects for improved diagnosis and control[J].The Journal of General Virolgy,2010,91(Pt12):2885-2897.
[3]MAJIYAGBE K A,NAWATHE D R,ABEGUNDE A.Rapid diagnosis of peste des petits ruminants(PPR) infection,application of immunoelectroosmophoresis(IEOP) technique[J].Rev Elev Med Vet Pays Trop,1984,37(1):11-15.
[4]SINGH R P,SREENIVASA B P,DHAR P,et al.Development of a monoclonal antibody based competitive-ELISA for detection and titration of antibodies to peste des petits ruminants(PPR) virus[J].Veterinary Microbiology,2004,98(1):3-15.
[5]SARAVANAN P,SINGH R P,BALAMURUGAN V,et al.Development of a N gene-based PCR-ELISA for detection of Peste-des-petits ruminants virus in clinical samples[J].Acta Virol,2004,48(4):249-255.
[6]BALAMURUGAN V,SEN A,SARAVANAN P,et al.One-step multiplex RT-PCR assay for the detection of peste des petits ruminants virus in clinical samples[J].Veterinary Research Communications,2006,30(6):655-666.
[7]BALAMURUGAN V,SEN A,VENKATESAN G,et al.Application of semi-quantitative M gene-based hydrolysis probe(TaqMan) real-time RT-PCR assay for the detection of Peste des petites ruminants virus in the clinical samples for investigation into clinical prevalence of disease[J].Transboundary and Emerging Diseases,2010,57(6):383-395.
[8]BATTEN C A,BANYARD A C,KING D P,et al.A real time RT-PCR assay for the specific detection of Peste des petits ruminants virus[J].Journal of Virological Methods,2011,171(2):401-404.
[9]ABERA T,THANGAVELU A,JOY CHANDRAN N D,et al.A SYBR Green I based real timeRT-PCR assay for specific detection and quantitation of Peste des petits ruminants virus[J].BMC Veterinary Research,2014,10:22.