五种重要猪病病毒基因芯片探针的建立及其应用

Development and application of gene chip for detection of five swine disease viruses

将猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪细小病毒(PPV)、猪伪狂犬病病毒(PRV)、猪圆环病毒2型(PCV-2)的特异cDNA片段作为探针点制于氨基修饰的玻片上,以这5种细胞毒的核酸作为模板,进行不对称PCR扩增,制备靶物,经绿色荧光染料Cy3标记后,分别与cDNA微阵列的芯片探针进行杂交,杂交信号经Genepix 4000A扫描仪扫描。结果显示,该芯片探针可以特异地与Cy3标记的这5种靶物结合。用该基因芯片对100份现地采集的猪全血样品进行检测,检出PRRSV阳性样品26份,PCV-2阳性样品47份,PRRSV和PCV-2混合感染阳性样品17份,PPV阳性样品5份,PRV阳性样品2份,CSFV阳性样品20份。表明,本试验研制的cDNA芯片探针可以特异地检测这5种猪病病毒,具有良好的诊断应用前景。

The specific cDNA probes of porcine circovirus type 2（PCV-2）,porcine reproductive and respiratory syndrome virus（PRRSV）,classical swine fever virus（CSFV）,porcine parvovirus（PPV）,pseudo-rabies virus（PRV） were fixed on the amino-modified slides.Asymmetric PCR products of five nucleic acids were labeled with Cy3 as the target material,and then hybridized with the cDNA probes on the slides.The slides were scanned by Genepix 4000A scanner.In result,the cDNA on the slide chips could specifically bind to the target material.100 whole blood samples were detected by the gene chip,and 26 were positive for PRRSV,47 for PCV-2,17 for PRRSV and PCV-2,5 for PPV,2 for PRV and 20 for CSFV.The results showed that the probes of the cDNA chip could detect the five viruses specially and had the potential for the diagnostic application.