欧洲型猪繁殖与呼吸综合征病毒荧光RT-PCR检测方法的建立.pdfVIP

26 Anmi alHusbandry VeterinaryM edicine 2008 Vol40 No7 欧洲型猪繁殖与呼吸综合征病毒荧光 RTPCR 检测方法的建立 1, 2 2 1 * 鲁韦韦 , 唐泰山, 姜 平 , 2 2 张常印, 王凯民 ( 1. , 210095; 2. , 210001) : (PRRSV ) , E1 E6 2 PCR TaqMan, E1 E6 RTPCR , PRRSV, 10DNAcDNA PCR , ( CSFV ) (PRV ) 7, E 1 E6 RTPCR PRRSV, 616 216 PRRSV DNA, CSFV PRRSV E1E6 RTPCR, PRRSV : ; RTPCR; : S85265 : A : 2008) R ealtim e RTPCR assays for the detection of European strain of porcine reproductive and resp iratory syndrom e virus 1, 2 2 1* L W eiwei , TANG Taishan, JIANG Ping , 2 2 ZHANG Changyin, WANG Kaim in ( 1. Key Laboratory ofAnmi alDisease D iagnostics and Immunology, M inistry ofAgriculture, NanjingAgricultural niversity, Nanjing 210095, China; 2. Jiangsu EntryExit Inspection and Quarantine Bureau, Nanjing 210001, China) Abstract: Two pairs of realtmi e RTPCR prmi ers and probes, E1 and E6, were designed and synthesized based on the nucleotide se quences ofEuropean strain of porcine reproductive and respiratory syndrome virus ( PRRSV). A fter the reaction conditionswere optmi ized, E1 and E6 realtmi e RTPCRwere developed here for PRRSV. The maxmi aldetection lmi its ofE1 and E6 realtmi eRTPCRwere 616 and 216 copies of plasmidDNA containing the gene ofPRRSV, respectively. The high specificity of the wt omethodswas confirmed by detecting 7 viruses, such as classicalsw ine fever virus (CSFV)pseudorabiesvirus