School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.
Crop Science Department, Teagasc, Oak Park, Carlow, Ireland.
Anal Biochem. 2022 Apr 1;642:114526. doi: 10.1016/j.ab.2021.114526. Epub 2021 Dec 17.
Potato virus Y (PVY) is an abundant and damaging virus which reduces crop yield and marketability. Accurate detection of this economically important virus both in-field and in seed potatoes is considered essential in the control of PVY spread. Current detection methods are focused on immunodetection and PCR-based methods, however, identification of PVY through isothermal amplification is a promising avenue for developing accessible, on-site diagnostics with quick turnaround times. In this work, a rapid recombinase polymerase amplification assay was developed which could readily amplify PVY nucleic acids with good sensitivity and specificity. Additionally, this assay was shown to be capable of amplification directly from RNA in a one-step amplification process, without the need for prior reverse transcription. The assay was coupled with lateral flow technology to provide a rapid visual confirmation of amplification. This nucleic-acid lateral flow immunoassay could feasibly be employed in-field, or at any location where testing is required, to aid in the detection and control of PVY.
马铃薯 Y 病毒（PVY）是一种丰富且具有破坏性的病毒，会降低作物产量和市场价值。在田间和马铃薯种子中准确检测这种具有重要经济意义的病毒，被认为是控制 PVY 传播的关键。目前的检测方法主要集中在免疫检测和基于 PCR 的方法上，然而，通过等温扩增来识别 PVY 是开发具有快速周转时间的便捷、现场诊断方法的一个有前途的途径。在这项工作中，开发了一种快速重组酶聚合酶扩增检测方法，该方法可以灵敏且特异地扩增 PVY 核酸。此外，该检测方法能够直接从 RNA 一步扩增过程中进行扩增，无需事先进行逆转录。该检测方法与侧流技术相结合，可快速进行扩增的可视化确认。这种核酸侧向流动免疫分析可以在田间或任何需要检测的地方使用，以帮助检测和控制 PVY。
