牛病毒性腹泻高效纳米PCR检测方法的建立及初步应用  

Development and primary application of a nanoparticle-assisted PCR assay for detection of bovine viral diarrhea disease virus

作　　者：李佳暖 李得鑫 崔元[1] 袁万哲[1,2] 孙继国 

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]河北省兽医生物工程技术研究中心,河北保定071001

出　　处：《中国兽医学报》2018年第2期252-254,共3页Chinese Journal of Veterinary Science

摘　　要：为建立快速、灵敏的牛病毒性腹泻病毒（BVDV）的检测方法，根据GenBank已发表的序列，在其5’端非结构蛋白区域设计了1对特异性引物，用来检测BVDV，PEDV，SRV，TGEV等，同时建立了牛病毒性腹泻纳米PCR检测方法，经优化找出最佳的NanoPCR反应条件。经过检测，纳米PCR的最小检出量为15．2×10^-4，PCR的最小检出量为15．2×10^-3；BVDV的敏感性试验表明纳米PCR方法的敏感性是普通PCR的10倍。该BVDV纳米PCR具有较高的敏感性和特异性，是一种高效的检测手段，可应用于流行病学调查，同时也为BVDV的及时检测、预防和治疗奠定基础。In order to develpop a rapid and sensitive assay for detection of BVDV,a pair of amplified 222 bp fragment specific primers to detect BVDV, PEDV, SRV, TGEV, etc based on the 5＇-un- translated region gene of bovine viral diarrhea virus（BVDV）. At the same time,developing a nano- particle assisted PCR assay for BVDV and finding the best reaction condition of a nanoparticle-as- sisted PCR. According the result, the minimum detectable amount of nanoparticle-assisted PCR was 15.2 × 10^- 4 ,the minimum detectable amount of PCR was 15.2 × 10^-3 ,the sensitivity of the nanoparticle-assisted PCR was 10 times more than the normal PCR. The BVDV nanoparticle-assis- ted PCR with high sensitivity and specificity,is a highly effective means of detection,it can be ap plied to epidemiological survey, but also lay a foundation for the timely detection, prevention and treatment of BVDV.

关 键 词：牛病毒性腹泻病毒(BVDV) 纳米PCR 5’端非结构蛋白 

分 类 号：S852.65[农业科学—基础兽医学]