Ivanov Aleksandr V, Safenkova Irina V, Zherdev Anatoly V, Dzantiev Boris B
A.N. Bach Institute of Biochemistry, Research Centre of Biotechnology, Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, Russia.
Plants (Basel). 2021 Nov 10;10(11):2424. doi: 10.3390/plants10112424.
Rapid, sensitive, and timely diagnostics are essential for protecting plants from pathogens. Commonly, PCR techniques are used in laboratories for highly sensitive detection of DNA/RNA from viral, viroid, bacterial, and fungal pathogens of plants. However, using PCR-based methods for in-field diagnostics is a challenge and sometimes nearly impossible. With the advent of isothermal amplification methods, which provide amplification of nucleic acids at a certain temperature and do not require thermocyclic equipment, going beyond the laboratory has become a reality for molecular diagnostics. The amplification stage ceases to be limited by time and instruments. Challenges to solve involve finding suitable approaches for rapid and user-friendly plant preparation and detection of amplicons after amplification. Here, we summarize approaches for in-field diagnostics of phytopathogens based on different types of isothermal amplification and discuss their advantages and disadvantages. In this review, we consider a combination of isothermal amplification methods with extraction and detection methods compatible with in-field phytodiagnostics. Molecular diagnostics in out-of-lab conditions are of particular importance for protecting against viral, bacterial, and fungal phytopathogens in order to quickly prevent and control the spread of disease. We believe that the development of rapid, sensitive, and equipment-free nucleic acid detection methods is the future of phytodiagnostics, and its benefits are already visible.
快速、灵敏且及时的诊断对于保护植物免受病原体侵害至关重要。通常，实验室中使用聚合酶链式反应（PCR）技术对植物的病毒、类病毒、细菌和真菌病原体的DNA/RNA进行高灵敏度检测。然而，使用基于PCR的方法进行现场诊断是一项挑战，有时几乎是不可能的。随着等温扩增方法的出现，该方法可在一定温度下扩增核酸且不需要热循环设备，分子诊断走出实验室已成为现实。扩增阶段不再受时间和仪器的限制。需要解决的挑战包括找到快速且用户友好的植物制备方法以及扩增后检测扩增子的合适方法。在此，我们总结基于不同类型等温扩增的植物病原体现场诊断方法，并讨论它们的优缺点。在本综述中，我们考虑等温扩增方法与适用于现场植物诊断的提取和检测方法的结合。在实验室外条件下进行分子诊断对于预防病毒、细菌和真菌植物病原体尤为重要，以便快速预防和控制疾病传播。我们相信，开发快速、灵敏且无需设备的核酸检测方法是植物诊断的未来，其益处已经显现。
