猪圆环病毒多重PCR诊断方法的建立

The establishment of a multiplex PCR of porcine circovirus

根据GenBank中20株猪圆环病毒(PCV2)核苷酸序列设计两对引物,建立的检测猪圆环病毒的多重PCR方法,其中引物P1、P2为PCV特异性,扩增938bp片段。P3、P4为PCV2特异性扩增490bp,因此本方法不仅可以扩增PCV片段,还可以同时区分PCV1和PCV2。该方法具有良好的特异性和敏感性,为猪圆环病毒的临床诊断与流行病学调查等研究奠定了基础。

The nuclear sequence homology and diversity of 20 strains of PCVI and PCV2 are compared to design two pairs of type-specific primers. The primers of Pl and P2 amplified the PCV virus-specific segment with the size of 938 bp. The primers of P3 and P4 amplified the PCV2 type-specific segment with the size of 490 bp. A multiplex PCR is developed based on these two pairs of primers to type and differentiate PCVl and PCV2.The optimization of the PCR reaction make the method more efficiently. The technique has been applied into the clinical diagnosis and epidemiological research.