FRRSV和PCV-2双重PCR检测方法的建立及应用

根据猪繁殖与呼吸综合征病毒(PRRSV)美洲型标准株(ATCCVR-2332)的ORF7保守序列和猪圆环病毒2型(PCV-2)(AF381175)的ORF2基因保守序列,设计合成了两对特异性引物。用这两对引物,通过优化的PCR条件,对PRRSV阳性毒株反转录后的cDNA模板和PCV-2毒株的DNA模板进行双重PCR扩增,同时得到两条与试验设计相符的432bp(PRRSV)和630bp(PCV-2)特异性条带,建立了同时检测PRRSV和PCV-2的双重PCR方法。并用此方法对在安徽省不同地区所采集的72头份病猪的淋巴结、肺、肝、脾、肾等组织进行检测,证明建立的PCR方法可用于临床诊断。

Establishment and Application of the Diplex PCR to the Detection of PRRSV and PCV-2

Based on conserved sequences in ORF7 fragments of PRRSV American strain(ATCCVR-2332) and that in ORF2 of PCV-2(AF381175) strain, two pairs of specific primers were designed respectively. Through optimized PCR conditions,the cDNA reversed from of PRRSV and the DNA of PCV-2 were amplify by the diplex PCR, two special fragments which were 432 bp(PRRV) and 630 bp(PCV-2) respectively. The diplex PCR for detection of PRRSV and PCV-2 was established.Seventy-two tissue samples of sick pigs(lymphocyte, lungs, liver, spleen and kidney)which from different areas of Anhui province were detected respectively by the diplex PCR,which indicated that the diplex PCR for detecting PRRSV and PCV-2 was a rapid,specific and sensitive,and could be used in clinic diganoses of the 2 diseases.