Moon Ye-Ji, Lee So-Young, Kim Unji, Oh Se-Wook
Department of Food and Nutrition, Kookmin University, Seoul, 02727 Republic of Korea.
Food Sci Biotechnol. 2023 May 8;33(1):203-209. doi: 10.1007/s10068-023-01315-z. eCollection 2024 Jan.
causing soft-rot disease requires on-site detection before the distribution of agricultural products. Loop-mediated isothermal amplification (LAMP), which is resistant to food inhibitors, is known for its high detection sensitivity for pathogens and when coupled with lateral flow immunoassay (LFA) enables visualizations. For detection of soft-rot disease, we developed a LAMP-LFA system targeting 16S ribosomal RNA, a partial sequence gene of subsp. . The LAMP-LFA was performed at 60 °C for 50 min followed by hybridization of digoxygenin-labeled LAMP amplicon and biotinylated probe. Detection sensitivity was 3.22 × 10 CFU/mL in pure culture, which specifically detected the target. In Chinese cabbage and potato, the target was detected up to low levels of 1.57 × 10 CFU/g and 1.29 × 10 CFU/g, respectively. This study showed potential applicability as a sensitive point-of-care system for soft-rot disease bacteria detection in agricultural products.
The online version contains supplementary material available at 10.1007/s10068-023-01315-z.
在农产品分发之前,对引起软腐病的病菌进行现场检测很有必要。环介导等温扩增技术(LAMP)对食品抑制剂具有抗性,以其对病原体的高检测灵敏度而闻名,当与侧向流动免疫分析(LFA)结合时能够实现可视化检测。为了检测软腐病,我们开发了一种针对亚种部分序列基因16S核糖体RNA的LAMP-LFA系统。LAMP-LFA在60℃下进行50分钟,随后进行地高辛标记的LAMP扩增子与生物素化探针的杂交。在纯培养物中检测灵敏度为3.22×10CFU/mL,能特异性检测目标物。在大白菜和马铃薯中,分别能检测到低至1.57×10CFU/g和1.29×10CFU/g的目标物。本研究表明该系统作为一种用于农产品中软腐病菌检测的灵敏即时检测系统具有潜在的适用性。
在线版本包含可在10.1007/s10068-023-01315-z获取的补充材料。