中国兽医科学2018，Vol.48Issue(5)：580-585,6.DOI:10.16656/j.issn.1673-4696.2018.0110
牛病毒性腹泻病毒Nano-PCR检测方法的建立及初步应用
Establishment and preliminary application of Nano-PCR for the detection of bovine viral diarrhea virus
摘要
Abstract
In order to establish a specific,sensitive and rapid molecular detection method for bovine viral diarrhea virus (BVDV),in this study,a novel nano-PCR for BVDV was established based on the combination of gold nanoparticle and traditional PCR technology with primers designed for BVDV 5'UTR conserved sequences. A comparative study of clinical samples was performed on the results of RT-PCR and the commercial IDEXX BVDV antigen Test Kit. The specificity and sensitivity tests showed that the Nano-PCR was only sensitive to BVDV,without cross reaction to the other viruses,such as infectious bovine rhinotracheitis virus,bovine respiratory syncytial virus and bovine parainfluenza type-3 virus. In the sensitivity test,the Nano-PCR showed 10 times sensitivity than that of the traditional PCR technology,with the minimum detection of 6.84×103 copies/ μ L. The clinical test results showed that the coincidence rate of this technique could reach to 100％. The positive detection rate of 38 clinical samples was 34.21％,which was 2.6 times that of RT-PCR and also the 1.5 times of the IDEXX kit. In conclusion,the established BVDV Nano-PCR can be used for the rapid diagnosis and identification of clinical samples, and has the advantages of high specificity,good sensitivity,fast and stable.关键词
牛病毒性腹泻病毒/Nano-PCR/RT-PCR/IDEXX试剂盒Key words
bovine viral diarrhea virus/Nano-PCR/RT-PCR/IDEXX kit分类
农业科技引用本文复制引用
李健友,刘泽余,刘占悝,李家伟,张淑琴,王超,郭利..牛病毒性腹泻病毒Nano-PCR检测方法的建立及初步应用[J].中国兽医科学,2018,48(5):580-585,6.基金项目
吉林省科技厅产业技术创新战略联盟项目(20170309002NY) （20170309002NY）
国家重点研发计划项目(2016YFD0500907) （2016YFD0500907）
中国农业科学院"创新工程"项目 （）
吉林省科技厅重点科技成果转化项目(20150307024NY) （20150307024NY）
