Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Wuhan 430070, China; College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Food Res Int. 2023 Jan;163:112212. doi: 10.1016/j.foodres.2022.112212. Epub 2022 Nov 25.
Rapid and sensitive quantitative detection methods are required to monitor and detect Salmonella throughout the food supply chain and early prevention of foodborne disease outbreaks. In this study, a magnetic microbead enzyme-linked immunoassay (MELISA) based on phage receptor binding protein was developed for rapid enrichment and detection of Salmonella in complex food matrices. RBP 41 from phage T102 acted as a species-specific recognition element for Salmonella by exploiting its strong binding capacity to Salmonella surface receptors. RBP 41-MBs were prepared by coupling recombinant RBP 41 with MBs and used to separate and enrich Salmonella cells from spiked food samples. The captured complexes were further integrated with ELISA procedures by HRP-labeled anti-Salmonella antibody for rapid and accurate detection of Salmonella. The whole method took <1.5 h and the detection limit was 10 CFU/mL. Therefore, MELISA was successfully developed for the detection of Salmonella in various spiked food samples (skim milk, lettuce, and chicken breast). The ELISA reaction process of this method was carried out on magnetic beads. It simplified the process of the traditional ELISA method and reduces the reaction time. This study expanded the use of phage-associated proteins and demonstrated the promising prospects for practical applications in the detection of foodborne pathogens.
需要快速和敏感的定量检测方法来监测和检测整个食物链中的沙门氏菌,并及早预防食源性疾病的爆发。在本研究中,基于噬菌体受体结合蛋白开发了一种磁性微球酶联免疫吸附测定(MELISA),用于快速富集和检测复杂食品基质中的沙门氏菌。噬菌体 T102 的 RBP 41 充当了沙门氏菌的种特异性识别元件,利用其与沙门氏菌表面受体的强结合能力。通过将重组 RBP 41 与 MB 偶联制备了 RBP 41-MB,并用于从加标食品样品中分离和富集沙门氏菌细胞。捕获的复合物通过 HRP 标记的抗沙门氏菌抗体进一步与 ELISA 程序集成,用于快速准确地检测沙门氏菌。整个方法耗时<1.5 小时,检测限为 10 CFU/mL。因此,成功地开发了 MELISA 用于检测各种加标食品样品(脱脂牛奶、生菜和鸡胸肉)中的沙门氏菌。该方法的 ELISA 反应过程在磁性珠上进行。它简化了传统 ELISA 方法的过程,并缩短了反应时间。本研究扩展了噬菌体相关蛋白的用途,并展示了在食源性病原体检测中的实际应用的广阔前景。