【摘要】 为建立快速、灵敏的猪细小病毒(PPV)的检测方法,本研究针对PPV VP2基因的保守区域设计一对扩增472 bp片段的特异性引物,建立了PPV纳米PCR检测方法。敏感性试验表明纳米PCR方法是普通PCR的1 000倍,最低核酸检出量为4拷贝/μL。利用该纳米PCR方法检测猪的其他相关病毒核酸,结果均呈阴性。分别采用纳米PCR和普通PCR对3个省份送检的43份临床样品进行检测,PPV阳性率分别为95%(41/43)和72%(31/41),表明该纳米PCR检测方法具有更高的敏感性,适用于PPV低含量临床样品的检测。更多还原

【Abstract】 To establish a rapid and sensitive nanoparticle-assisted PCR(nanoPCR) assay for detection of porcine parvovirus(PPV), primers were designed based on the conserved region of the VP2 gene to amplify a 472 bp fragment. The recombinant plasmid of pMD-VP2 which contained full-length VP2 gene was constructed. The sensitivity tests of nanoPCR and conventional PCR showed that nanoPCR was 1 000 times more sensitive than conventional PCR with the diluted pMD-VP2 as the template. In addition, the other porcine viruses detected by the nanoPCR were all negative. Furthermore, the PPV positive rates of 95% and 72% were detected from 43 clinical samples collected from three provinces in China by the nanoPCR and conventional PCR, respectively. These data indicated that the nanoPCR is more sensitive and applicable for detection of PPV infection in pigs.更多还原