摘要
根据GenBank上已发表的猪伪狂犬病毒(PRV)的gH基因、猪细小病毒(PPV)的VP2基因序列和猪圆环病毒2型(PCV2)的ORF2基因序列,设计合成了3对特异引物,分别建立了PRV,PPV和PCV2的单项PCR诊断方法;通过对扩增条件的筛选,建立了PRV,PPV,PCV2的复合PCR诊断方法,利用1次PCR反应,即可同时扩增PRV的355 bp,PPV的195 bp和PCV2 487 bp的特异性片段,而猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、正常细胞(PK-15)扩增结果均为阴性,对PRV,PPV,PCV2的最低检出量分别为100 fg,1 pg和10 pg的DNA.该方法适合对PRV,PPV和PCV2的联合检测和鉴别诊断.
On the basis of gH sequences of PRV,VP2 sequences of PPV and ORF2 sequences of PCV2 obtained from the GenBank,three pairs of primers were designed,single PCR of PRV,PPV and PCV2 were established.The Multiplex PCR was developed by optimized conditions.The results showed that 355 bp for PRV,195 bp for PPV and 487 bp for PCV2 could be amplified in a multiplex PCR.DNA fragments were not amplified from HCV,PRRSV and control cells（PK-15）,and it could detect the template DNA of 100 fg for PRV,1 pg for PPV and 10 pg for PCV2,and it could detect PRV,PPV,PCV2 in a mixed infection and identification diagnosis.
出处
《河南农业大学学报》
CAS
CSCD
北大核心
2010年第4期421-424,共4页
Journal of Henan Agricultural University
基金
河南省重点科技攻关项目(08210213007)
国家质检局项目(2009IK029)
关键词
多重PCR
检测
猪伪狂犬病毒
猪细小病毒
猪圆环病毒2型
multiplex PCR
detection
porcine pseudorabies virus
porcine parvovirus
porcine circovirus type 2
