猪圆环病毒2型地高辛标记探针检测方法的建立与应用  

Establishment and application of digoxigenin-labeled nucleic acid probe for PCV-2

作　　者：刘方娜[1] 刁有祥[1] 王妮[1] 孟凡磊[1] 王辉[1] 张伟[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2008年第7期771-774,共4页Chinese Journal of Veterinary Science

基　　金：山东省重大畜禽疫病综合防治技术资助项目(011020102)

摘　　要：根据GenBank中已发表的猪圆环病毒2型(PCV-2)基因序列,在ORF2内设计1对特异性引物,利用PCR反应扩增出371 bp的核酸片段,回收并纯化PCR产物,用地高辛标记,建立了地高辛标记核酸探针诊断PCV-2的方法。该探针与8个PCV-2重组菌的核酸抽提物均能发生特异性杂交,而与对照猪圆环病毒Ⅰ型(PCV-1)、猪繁殖与呼吸综合征病毒(PRRSV)、猪细小病毒(PPV)、猪伪狂犬病病毒(PRV)的核酸杂交均为阴性;对PCV-2 DNA的最低检测量为10 pg。According to the genomic sequence of PCV-2 published in GenBank,one pair of primers were designed in the ORF2 for amplifying the 371 bp fragment in PCR experiments.The PCR product was labeled with digoxigenin as DNA probe for detection of PCV-2.The hybridization assay result showed that the eight recombinant bacteria of PCV-2 were positive,but no cross-hybridization was detected with DNAs of PCV-1,PRRSV,PPV and PRV.The sensitivity result indicated that as few as 10 pg DNA of PCV-2 could be detected by the D...

关 键 词：地高辛标记核酸探针 PCV-2 ORF2 

分 类 号：S852.65[农业科学—基础兽医学]