Development and Application of SYBR Green Ⅰ Fluorescent Quantitative PCR for Detection of Cap Gene of Pigeon Circovirus
In order to develop a rapid diagnostic method for pigeon circovirus(PiCV),a pair of specific primers were designed and synthesized according to the cap conservative gene sequence of PiCV for development of a fluorescence quantitative PCR method.Subsequently,this assay was evaluated for its sensitivity,specificity,repeatability and suitability for testing clinical samples.The results showed that under the optimal reaction conditions,this assay had a good linear correlation in the range of 1×102-1×107 copies/μL and the linear correlation coefficient of 0.9846.The minimum detection limit of standard substance was 1 × 102 copies/μL,which was 100 times sensitive than that of the conventional PCR method.The method did not react with other common pigeon pathogens.Repeatability evaluation results showed that the coefficients of variation within and between batches were less than 2％.Then this method along with the conventional PCR was used to test liver samples of dead pigeons.The positive rates of PiCV were 75.7％for the fluorescence quantitative PCR method and 54.1％for conventional PCR.Taken together,this method developed in this study could be used as a better choice for rapid detection of PiCV and technical support for the timely prevention and control of PiCV.
Cap genepigeon circovirusreal-time quantitative PCR
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