江西农业大学学报2018，Vol.40Issue(1)：136-141,6.DOI:10.13836/j.jjau.2018018
新现猪圆环病毒3型及圆环病毒2型双重PCR检测方法的建立与应用
Establishment and Application of Double PCR Assays for Detection of New-emerged Porcine Circovirus Type 3(PCV-3) and Porcine Circovirus Type 2(PCV-2)
摘要
Abstract
According to the published ORF2 gene sequences of new emerged porcine circovirus 3(PCV-3) and porcine circovirus 2(PCV-2)in GenBank,two pairs of primers specific to PCV-3 and PCV-2 were designed and synthesized.By optimizing the reaction conditions,duplex polymerase chain reaction(PCR)-based assay was developed and evaluated for its ability to simultaneously detect co-viral infections of swine.The detection limit of the established assay was 1.0 x103copies/μL for both PCV-3 and PCV-2.When common swine viral pathogens,including pseudorabies virus(PRV),porcine deltacoronavirus(PDCoV),porcine epidemic diarrhea virus(PEDV),porcine transmissible gastroenteritis virus(TGEV),porcine reproductive and respiratory syn-drome virus(PRRSV)and classical swine fever virus(CSFV),served as the template for the evaluation of the specificity of the assay,no cross amplification was observed.Based on the clinical investigation on 256 samples, the positive rates of PCV-3 and PCV-2 were 0.78%(2/256)and 86.3%(221/256),respectively.Only one positive sample was co-infected with PCV-3 and PCV-2.This study showed that the duplex PCR established in this study was sensitive and specific,and would be a useful tool for clinical diagnosis and epidemiologic survey of PCV-3 and PCV-2 circulating in swine.关键词
新现猪圆环病毒3型/猪圆环病毒2型/双重PCR/应用Key words
porcine circovirus type 3/porcine circovirus type 2/duplex PCR/application分类
农业科技引用本文复制引用
张帆帆,宋德平,唐玉新,李龙显,叶昱,李凯,郭楠楠,张敏,黄冬艳,吴琼,何后军..新现猪圆环病毒3型及圆环病毒2型双重PCR检测方法的建立与应用[J].江西农业大学学报,2018,40(1):136-141,6.基金项目
国家自然科学基金面上项目(31372457)和江西省教育厅科技计划项目(GJJ160399)Project supported by the National Natural Science Foundation of China(31372457)and Foundation of Education-al Commission of Jiangxi Province(GJJ160399) （31372457）
