Establishment And Application Of Real-time Quantitative PCR For Detection Of Pigeon Adenovirus Type Ⅰ

Pigeon adenovirus Type Ⅰ(PIAd Ⅴ-Ⅰ),as a contagious pathogen,has a wide range of transmission and high incidence,especially for racing pigeons,which are in a highly stressed state during training and become susceptible animals due to immunosuppression.Sporadic cases of PIAd Ⅴ-Ⅰ infection had been diagnosed in the 1970 s,but it was not until 1984 Cousement et al that the first description of PIAd Ⅴ-Ⅰ,a clinical disease found only in pigeons in Belgium,was made.Since then,PIAd Ⅴ-Ⅰ infection has not only been confined to a certain region,but has spread and been widely reported all over the world,and has brought a great impact on the healthy development of pigeon breeding industry.In this study,according to the Pi Ad Ⅴ-Ⅰ Fiber 2 gene sequence published by Gen Bank,Primer 5.0 was used to design specific amplification primers,and the PCR detection method for Pi Ad Ⅴ-Ⅰ was established by optimizing the reaction program and system.A967 bp Fiber 2 conserved gene sequence was obtained,which was identified as avian adenovirus TYPE A and type 1 after genetic evolution analysis.Based on this,Pi Ad Ⅴ-Ⅰ SYBR Green ⅰ fluorescence quantitative PCR method was established,and compared with conventional PCR by clinical sample detection,the results showed that real-time fluorescence quantitative PCR method was 100 times more sensitive than the conventional PCR method.