中国畜牧兽医2017，Vol.44Issue(12)：3440-3445,6.DOI:10.16431/j.cnki.1671-7236.2017.12.007
PCV2和PCV3双重纳米PCR方法的建立及初步应用
Establishment and Rudimentary Application of the nano-dPCR for Detection of PCV2 and PCV3
摘要
Abstract
This study was aimed to establish a duplex nanoparticle-assisted PCR (nano-dPCR) method for the simultaneous detection of porcine circovirus type 2 (PCV2) and PCV3,and to apply this method in the detection of PCV2 and PCV3.Primers specific to PCV2 and PCV3 were designed by reference to the respective gene sequences in GenBank.The reaction conditions of nano-dPCR were also optimized.An evaluation of the specificity and sensitivity of the established nano-dPCR protocol proved the method to be both specific and sensitive,with the lower detection limit for the amount of nucleic acid in PCV2 and PCV3 to be 93.2 and 91.6 copies/μL,respectively.This sensitivity was 100 times higher than that of conventional PCR.The method was applied to inspect 265 clinical samples sent for testing,and the results showed that PCV2 and PCV3 infection in pig was rather common,with 16.6％ positive for PCV3,14.7％ positive for PCV2,and 6.8％ for mixed infection.The detection results on clinical samples supported the newly established nano-dPCR method as a rapid and sensitive differential diagnosis for the early infection of PCV2 and PCV3.关键词
双重纳米PCR/猪圆环病毒2型(PCV2)/猪圆环病毒3型(PCV3)/病原检测Key words
duplex nanoparticle-assisted PCR (nano-dPCR)/porcine circovirus 2 (PCV2)/porcine circovirus 3 (PCV3)/pathogen detection分类
农业科技引用本文复制引用
梁琳,逄春华,罗亚坤,周灵,王静,刘畅,刘琪,崔尚金..PCV2和PCV3双重纳米PCR方法的建立及初步应用[J].中国畜牧兽医,2017,44(12):3440-3445,6.基金项目
中国农业科学院基本科研业务费(2016ywf-yb-12) （2016ywf-yb-12）
中国农业科学院创新工程(ASTIP-IAS15) （ASTIP-IAS15）
