关键词: 猪瘟病毒；猪繁殖与呼吸综合征病毒；双重实时荧光定量RT\, PCR

Abstract: To identify classical swine fever virus（CSFV） and porcine reproductive and respiratory syndrome virus（PRRSV） rapidly， a double real\|time fluorescent RT\|PCR assay of CSFV and PRRSV was designed， along with primers and TaqMan probes based on CSFV and PRRSV genome sequences. It was shown that the proposed assay proved to be specific， as no amplification was found of PCV2，PRV，PPV，PEDV，JEV，BVDV. The assay was sensitive to as low as 20 copies of template RNA of PRRSV and CSFV， and the sensitivity was 10 times higher than that of the conventional RT\|PCR. The coefficient variation （CV%） of intra/inter\|assay for the same RNA sample was less than 4%. Fifty samples were examined by the proposed fluorescent quantitative RT\|PCR and the conventional RT\|PCR， respectively， and there were 30 and 13 samples found to be infected with PRRSV and CSFV， respectively， by the proposed assay， but only 24 and 10 samples were found to be infected with PRRSV and CSFV， respectively， by the conventional RT\|PCR. These results showed that the developed double real\|time fluorescent quantitative RT\|PCR assay was rapid， specific， sensitive and simple for the detection of PRRSV and CSFV， which could be used as a rapid detection method for the epidemiologic survey of PRRSV and CSFV.

Key words: classical swine fever virus（CSFV）, porcine reproductive and respiratory syndrome virus （PRRSV）, double real\, time fluorescent quantitative RT\, PCR