猪圆环病毒2型的PCR检测方法应用  

Application of PCR Detection for Porcine Circovirus Type 2

作　　者：苗丽娟[1,2,3] 李静姬 李一经[1] Miao Lijuan;Li Jingji;Li Yijing(Northeast Agricultural University,Harbin,150030;Jilin Agricultural Science and Technology College,Jilin,132101;Key Laboratory of Preven-tive Veterinary Medicine of Jilin Province,Jilin,132101)

机构地区：[1]东北农业大学,哈尔滨150030 [2]吉林农业科技学院,吉林132101 [3]预防兽医学吉林省重点实验室,吉林132101

出　　处：《基因组学与应用生物学》2019年第3期1383-1386,共4页Genomics and Applied Biology

基　　金：吉林省科技发展计划项目(20150623004TC);吉农院合字[(2015)第X057号]共同资助

摘　　要：本研究登录Genbank对猪圆环病毒2型基因的序列进行分析,用Primer 5.0设计了ORF2基因的扩增引物,试图选择一种比较合理的PCR方法检查PCV2感染的病原,以期这种PCR方法可以有效分析猪综合征障碍病毒(PRRSV)、猪细小病毒(PPV)、猪瘟病毒的扩增(HCV)、猪伪狂犬病毒(PRV)等比较常见的病原。研究结果表明,在PCV2进行扩增阳性样品检测中,发现一条异常447 bp的DNA条带,对产物测序结果进行扩增分析,证明其是PCV ORF2基因序列。敏感性检验分析表明检测样本DNA浓度时达到了9.8×10^(-4)ng/μL。PCR实验具有良好的稳定性和重复性。根据对66例临床病猪的分析表明,在PCV2的检测中阳性感染率是27.26%,这可能受到HCV、PRRSV、PRV、PPV等多种病毒的感染,其中混合感染的比例达到了72.23%。In this study,the sequences of porcine circovirus type 2 gene were analyzed by logging in Genbank,and the ORF2 gene amplification primers were designed with Primer 5.0.We tried to select a reasonable PCR method to examine the pathogen of PCV2 infection,hoping to effectively analyze the common pathogens such as porcine syndrome disorder virus(PRRSV),porcine parvovirus(PPV),hog cholera virus(HCV)amplification and porcine pseudorabies virus(PRV).The results showed that an anomaly DNA band of 477 bp was found in the detection of PCV2 amplified positive samples,and the product sequencing results were amplified to be proved that it was a PCV ORF2 gene sequence.Sensitivity analysis showed that the DNA concentration of the tested sample reached 9.8×10^-4 ng/μL;PCR had good stability and repeatability.According to the analysis of 66 clinical sick pigs,the positive infection rate in PCV2 test was 27.26%,which might be infected by HCV,PRRSV,PRV,PPV and other viruses,among which the proportion of mixed infection reached 72.23%.

关 键 词：猪圆环病毒2型 PCR检测 敏感性检验 

分 类 号：S852.651[农业科学—基础兽医学]