Establishment and Application of a Universal Real-time Fluorescent PCR for Pigeon Adenovirus
Pigeon adenovirus(PiAdV)is one of the major pathogens affecting healthy development of pigeon industry.In order to rapidly detect PiAdV at high-throughput,based on the common conserved region of pⅧ gene sequences of PiAdV-A and PiAdV-B registered in GeneBank,specific primers and probes were designed,after optimization of reaction conditions,a universal real-time fluorescent PCR assay for detecting PiAdV was established,followed by tests of its specificity,sensitivity and repeatability.The results showed that the optimal probe concentration of the established method was 0.2 μmol/L,without any cross reactivity with other common pigeon viruses,the minimum detection limit was 56.9 copies/μL,100 times more sensitive than conventional PCR,and all the coefficients of variation in repeated inter and intra groups were less than 1.8％.20 suspected samples were detected by the method and the conventional PCR,the detection rate of positive samples by the former was higher than that by the latter.In conclusion,the universal real-time fluorescent PCR assay for PiAdV established in the paper,with its advantages of strong specificity,high sensitivity and good reliability,could be used for clinical detection and epidemiological investigation of the virus.
万方数据
维普
