Beijing Laboratory of Food Quality and Safety, Beijing Key Laboratory of Agricultural Product Detection and Control of Spoilage Organisms and Pesticide Residue, Beijing Engineering Technology Research Center of Food Safety Immune Rapid Detection, College of Food Science and Engineering, Beijing University of Agriculture, Beijing, China.
Department of Nutrition and Health, Key Laboratory of Functional Dairy, Co-constructed by Ministry of Education and Beijing Government, China Agricultural University, Beijing, China.
PLoS One. 2022 Dec 12;17(12):e0278869. doi: 10.1371/journal.pone.0278869. eCollection 2022.
Shigella spp. and enteroinvasive Escherichia coli (EIEC) are widely distributed and can cause serious food-borne diseases for humans such as dysentery. Therefore, an efficient detection platform is needed to detect Shigella and EIEC quickly and sensitively. In this study, a method called recombinase polymerase amplification combined with lateral flow dipstick (RPA-LFD) was developed for rapid detection of Shigella and EIEC. RPA primers and LFD detection probes were designed for their shared virulence gene ipaH. Primers and probes were screened, and the primer concentration, and reaction time and temperature were optimized. According to the optimization results, the RPA reaction should be performed at 39°C, and when combined with LFD, it takes less than 25 min for detection with the naked eye. The developed RPA-LFD method specifically targets gene ipaH and has no cross-reactivity with other common food-borne pathogens. In addition, the minimum detection limit of RPA-LFD is 1.29×102 copies/μL. The detection of food sample showed that the RPA-LFD method was also verified for the detection of actual samples.
志贺氏菌属和侵袭性大肠杆菌（EIEC）分布广泛，可引起严重的食源性疾病，如痢疾。因此，需要一种高效的检测平台来快速、灵敏地检测志贺氏菌属和 EIEC。在本研究中，开发了一种称为重组聚合酶扩增结合侧流试纸条（RPA-LFD）的方法，用于快速检测志贺氏菌属和 EIEC。针对其共同的毒力基因 ipaH 设计了 RPA 引物和 LFD 检测探针。筛选了引物和探针，并优化了引物浓度、反应时间和温度。根据优化结果，RPA 反应应在 39°C 下进行，与 LFD 结合时，肉眼检测不到 25 分钟。开发的 RPA-LFD 方法特异性靶向基因 ipaH，与其他常见食源性病原体无交叉反应。此外，RPA-LFD 的最小检测限为 1.29×102 拷贝/μL。对食品样本的检测表明，RPA-LFD 方法也可用于实际样本的检测。
