【摘要】 为建立猪瘟病毒(CSFV)野毒株和疫苗株快速定量鉴别检测方法,根据CSFV野毒株和疫苗株E2基因序列差异,设计了2对特异性引物及1条Taq Man探针,以含CSFV HB株和C-株E2基因的重组质粒p BS-E2C和p BS-E2作为标准品,建立了能同时检测CSFV野毒株和疫苗株的双重Taq Man real-time PCR鉴别检测方法。结果表明,建立的CSFV双重Taq Man real-time PCR标准曲线Ct值与1×101~1×106copies/μL之间的E2基因拷贝数呈良好的线性关系,灵敏度达10 copies/μL,且特异性和重复性很好。综上,本试验建立的Taq Man real-time PCR检测方法可用于CSFV野毒株和疫苗株的鉴别诊断及病原定量分析。更多还原

【Abstract】 This experiment was conducted to establish a duplex Taq Man real-time PCR for differentiating C-strain vaccine and wild-type viruses of CSFV. According to the differences of the E2 gene sequences between C-strain and wild-type of CSFV,two pairs of specific primers and a Taq Man probes were designed in the E2 gene. The recombinant plasmid,which contained E2 gene of C-strain vaccine and wild-type viruses of CSFV was used to establish melting and standard curves of the duplex Taq Man quantitative real-time PCR. The standard curve for the viral genomic copy number and threshold cycle ranging from 1 × 101-1 ×106copies/μL E2 gene were linear. Sensitivity of the method was 10 copies,with a good specificity and repeatability. These data indicated that the duplex Taq Man quantitative real-time PCR can be used to CSFV diagnosis and quantification.更多还原