DEVELOPMENT OF A QUANTITATIVE REAL-TIME PCR FOR DETECTION OF IFN-α MRNA OF MUSCOVY DUCKS

Two pairs of primers were designed according to the duck IFN-α and β-actin gene sequences available in GenBank for the development of a SYBR Green I quantitative Real-time PCR.The β-actin was used as an internal control for the assay.To establish the standard curve,positive plasmid of each gene served as a standard and the melting curve was also analyzed.The results showed a good linear correlation coefficient(r0.99) as Ct values ranged from 13.27 to 27.54 for IFN-α and from 11.86 to 25.32 for β-actin.The melting curves presented single peaks with(93.6±0.26)℃ and(85.3±0.15)℃,respectively.The detection limits were 9.37×102 copies/μL for IFN-α and 3.71×101copies/μL for β-actin.The Real-time PCR developed in the present study could detect IFN-α at high efficiency within 4 hours.Therefore,this assay could be used for quantitative analysis of IFN-α mRNA expression in Muscovy ducks.