College of Animal Science and Technology, Shandong Agricultural University, Tai'an, Shandong, 271018, China.
College of Animal Science and Technology, Shandong Agricultural University, Tai'an, Shandong, 271018, China.
Poult Sci. 2020 Dec;99(12):6446-6453. doi: 10.1016/j.psj.2020.08.021. Epub 2020 Aug 26.
Fowl adenovirus (FAdV) has posed a grave threat to the health of poultry, and the sudden outbreak highlights the importance of the new rapid diagnostic method for the control and prevention of transmission. Hence, in the present study, a novel recombinase polymerase amplification (RPA) assay, which was suitable for all 12 serotypes (FAdV-1 to 8a and 8b to 11) had been successfully launched to detect FAdV. Also, the entire amplification process could be completed in the isothermal condition when temperature ranged from 26 to 42°C within no more than 14 min, which was remarkably superior to endpoint polymerase chain reaction (98 min) with the same detecting sensitivity (as low as 0.1 fg viral DNA), avoiding sophisticated thermal cyclers with simple operation. Additionally, the same primers did not produce positive reactions with other viruses tested, demonstrating that the specificity of the RPA assay was acceptable. Moreover, this developed method could be efficiently used in the diagnosis of FAdV references and epidemic strains from different avian origins, thus making it a rapid, reliable, and point-of-care FAdV diagnostics tool, as well as an alternative to endpoint PCR.
禽腺病毒（FAdV）对家禽的健康构成了严重威胁，其突发疫情凸显了新的快速诊断方法对于控制和预防传播的重要性。因此，在本研究中，我们成功开发了一种适用于所有 12 种血清型（FAdV-1 至 8a 和 8b 至 11）的新型重组酶聚合酶扩增（RPA）检测方法，用于检测 FAdV。此外，在 26 至 42°C 的等温条件下，整个扩增过程可在 14 分钟内完成，这明显优于具有相同检测灵敏度（低至 0.1 fg 病毒 DNA）的终点聚合酶链反应（98 分钟），避免了使用复杂的热循环仪进行简单的操作。此外，相同的引物不会与其他测试的病毒产生阳性反应，表明 RPA 检测方法的特异性是可以接受的。此外，该方法可有效用于不同禽类来源的 FAdV 参考株和流行株的诊断，因此，它是一种快速、可靠的即时检测 FAdV 的诊断工具，可作为终点 PCR 的替代方法。
