Adenoviruses representative strain information.

Pigeon virus diarrhea is very common in the clinic, which has caused great damage to the pigeon industry. Pigeon adenovirus constitutes a significant pathogen responsible for pigeon viral diarrhea. Among them, Pigeon adenovirus 1 (PiAdV-1) and Pigeon adenovirus 2 (PiAdV-2) are the most common adenoviruses discovered in pigeon diseases in recent years. Infection with these viruses is common in clinical settings, which makes differential diagnosis more difficult. At present, real-time PCR is one of the most widely used techniques for the identification and detection of pathogens. This study presents the development of a dual real-time PCR assay designed specifically for the detection of PiAdV-1 and PiAdV-2. The dual real-time PCR assay developed in this study demonstrated satisfactory specificity, sensitivity, repeatability, and reproducibility, while showing no cross-reactivity with unrelated pathogens. The limit of detection (LOD) for PiAdV-1 is 94.8 copies/µL, and the LOD for PiAdV-2 is 88.4 copies/µL. This method was then used to analyze 20 liver tissue samples collected from dead pigeons, and the results were verified by genome sequencing. To understand the current epidemic status of pigeon adenovirus in China, the dual real-time PCR method established in this study was used to detect 500 pigeon swab samples collected from 10 provinces in China from 2022 to 2023. The results showed that the positive rates of PiAdV detected by the real-time PCR was 27.00% (135/500). PiAdV-1 single infection was detected in 8 cases (1.6%), PiAdV-2 single infection in 123 cases (24.6%), and PiAdV-1 and PiAdV-2 co-infection in 4 cases (0.8%). This study established a valuable tool for differentiating between PiAdV-1 and PiAdV-2 in practical applications and provided significant insights into the prevalence of these viruses in China.

鸽病毒性腹泻在临床中极为常见，已对鸽养殖业造成了严重危害。鸽腺病毒是引发鸽病毒性腹泻的重要病原之一，其中，鸽腺病毒1型（Pigeon adenovirus 1, PiAdV-1）与鸽腺病毒2型（Pigeon adenovirus 2, PiAdV-2）是近年在鸽病中检出的最常见腺病毒毒株。该类病毒感染在临床中多发，这使得鉴别诊断难度大幅提升。目前，实时荧光定量PCR（real-time PCR）是应用最广泛的病原鉴定与检测技术之一。本研究开发了一种专为检测PiAdV-1与PiAdV-2设计的双重实时荧光定量PCR检测方法。该方法展现出良好的特异性、灵敏度、重复性与再现性，且与其他非相关病原无交叉反应。其对PiAdV-1的检测限（limit of detection, LOD）为94.8 copies/µL，对PiAdV-2的检测限为88.4 copies/µL。随后，研究人员使用该方法对20份病死鸽肝脏组织样本进行检测，检测结果通过基因组测序得以验证。为明确我国鸽腺病毒的当前流行态势，本研究使用所建立的双重实时荧光定量PCR方法，对2022至2023年间从国内10个省份采集的500份鸽拭子样本进行了检测。结果显示，经实时荧光定量PCR检测，鸽腺病毒总体阳性率为27.00%（135/500）。其中，单感染PiAdV-1的样本有8例（占比1.6%），单感染PiAdV-2的样本有123例（占比24.6%），同时感染PiAdV-1与PiAdV-2的样本有4例（占比0.8%）。本研究为临床实际应用中区分PiAdV-1与PiAdV-2提供了极具价值的检测工具，并为明确我国该类病毒的流行情况提供了重要参考。