由副溶血性弧菌引起的食品中毒事件频繁爆出，开发一种简单、快速且能大规模现场检测的方法对确保食品安全、避免经济损失具有重要意义。该研究成功开发了针对副溶血性弧菌现场快速检测的时间分辨荧光免疫层析技术（TRF-LFIA）。该方法将时间分辨纳米荧光微球（EuNPs）标记副溶血性弧菌单克隆抗体形成可以作为特异性免疫荧光探针的微球抗体偶联物，通过测定探针与目标致病菌结合后的荧光信号值来实现对目标致病菌的定量检测。该研究开发的TRF-LFIA方法对副溶血性弧菌检测的灵敏度为8.2×102 CFU/mL，视觉检测限为1.2×104 CFU/mL，线性范围为1.8×103~1.8×107 CFU/mL，回收率为84.25%~105.13%，变异系数（CV）为2.56%~9.14%，并且对其他7种主要食源性致病菌株检测未发生交叉反应。该研究所建立的TRF-LFIA检测方法具有灵敏度高，操作简单、快速，重复性、特异性好等优点，为针对副溶血性弧菌的现场快速检测提供了一种有力的检测工具。

Vibrio parahaemolyticus-induced food poisoning is frequently reported, highlighting the necessity to develop a simple, rapid, large-scale on-site detection method to ensure food safety and avoid financial losses. Time-resolved fluorescence lateral flow immunoassay (TRF-LFIA) was successfully developed for the rapid detection of Vibrio parahaemolyticus, using europium (III) nanoparticles (EuNPs) to label V. parahaemolyticus monoclonal antibodies, forming microsphere-antibody conjugations for use as specific immunofluorescent probes. The intensity of fluorescence signals emitted by the combination of the probes and the target pathogenic bacteria could be detected to achieve quantitative detection of the target pathogenic bacteria. The TRF-LFIA has a sensitivity of 8.2×102 CFU/mL, a visual detection limit of 1.2×104 CFU/mL, a linear range of 1.8×103~1.8×107 CFU/mL, a recovery rate range of 84.25%~105.13%, and a coefficient of the variation range of 2.56%~9.14%. There was no cross-reaction to the other seven major foodborne pathogenic strains. The TRF-LFIA established in this study has high sensitivity, allowing for simple and rapid operation, and good repeatability and specificity, providing a powerful detection tool for rapid on-site detection of V. parahaemolyticus.

广东省基础与应用基础研究项目（2020A1515011561）