Dickeya solani, one of the most significant bacterial pathogens, infects potato plants, resulting in severe economic damage. In this study, a lateral flow assay (LFA) combined with isothermal DNA amplification was developed for rapid, specific, and sensitive diagnosis of the potato blackleg disease caused by D. solani. Recombinase polymerase amplification (RPA) was chosen for this purpose. Five primer pairs specific to different regions of the D. solani genome were designed and screened. A primer pair providing correct recognition of the target sequence was aligned with the SOL-C region specific to D. solani and flanked by fluorescein (forward primer) and biotin (reverse primer). Lateral flow test strips were constructed to detect DNA amplicons. The RPA-LFA demonstrated a detection limit equal to 14,000 D. solani colony-forming units per gram of potato tuber. This assay provided sensitivity corresponding to the polymerase chain reaction (PCR) but was implemented at a fixed temperature (39 °C) over 30 min. No unspecific reactions with Pectobacterium, Clavibacter, and other Dickeya species were observed. Detection of latent infection of D. solani in the potato tubers by the developed RPA-LFA was verified by PCR. The obtained results confirmed that RPA-LFA has great potential for highly sensitive detection of latent infection.

Dickeya solani是最重要的细菌病原体之一，可感染马铃薯，造成严重的经济损失。在这项研究中，开发了一种结合等温DNA扩增的侧向流动测定（LFA），用于快速，特异性和灵敏地诊断由茄D病引起的马铃薯黑腿病。为此选择了重组酶聚合酶扩增（RPA）。设计并筛选了对茄形梭菌基因组不同区域特异的五个引物对。提供正确识别靶序列的引物对与D. solani特异的SOL-C区对齐侧翼是荧光素（正向引物）和生物素（反向引物）。构建横向流动测试条以检测DNA扩增子。RPA-LFA的检出限等于每克马铃薯块茎14,000 D. solani菌落形成单位。该测定法提供了与聚合酶链反应（PCR）相对应的灵敏度，但在固定温度（39°C）下进行了30分钟。没有非特异性反应黑腐，棍状杆菌，和其他的Dickeya观察到的物种。茄形衣原体潜伏感染的检测PCR验证了开发的RPA-LFA在马铃薯块茎中的抗性。获得的结果证实，RPA-LFA具有高度灵敏的潜伏感染检测潜力。