根据Genbank上已发表的猪伪狂犬病毒(PRV)的gH基因和猪细小病毒(PPV)的VP2基因的保守序列,设计合成了2对特异引物,分别建立了PRV和PPV的单项PCR诊断方法,通过优化PCR条件最终成功地建立了PRV和PPV的复合PCR诊断方法.敏感性检测结果表明,对PRV的检测可以达到10-10.5/100 μL TCID50、对PPV的检测可以达到10-9.5/100 μL TCID50.对20头份病、死猪的血清、淋巴结、肺、肝等组织进行检测,结果5头份PRV阳性,3头份PPV阳性,其中3头份PRV和PPv同时为阳性,其余猪为阴性,健康猪对照样品全部阴性.结果表明,PRV和PPV复合PCR诊断方法具有高度特异性和敏感性,可用于兽医临床诊断.
According to sequences of PRV and PPV published in GenBank,two pairs of primers were designed,single PCR of PRV and PPV were established,The duplex PCR was developed by optimizing action conditions,This method could detect the template DNA of 10 -10.5/100 μL LTCID50 for PRV,and 10-9.5/100 μL LTCID50 for PPV.Twenty organ samples from different farms of Henan province were respectively detected by duplex PCR,out of which five were PRV positive,eight were PPV positive,three were both PRV and PPV positive,control samples were negative.It suggested that the duplex PCR of PRV and PPV was a rapid,specific and sensitive diagnostic method.
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