Ma Biao, Li Jiali, Chen Kai, Yu Xiaoping, Sun Chuanxin, Zhang Mingzhou
Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, China.
Department of Plant Biology, Uppsala BioCenter, Linnean Centre for Plant Biology, Swedish University of Agricultural Science (SLU), SE-75007 Uppsala, Sweden.
Foods. 2020 Mar 3;9(3):278. doi: 10.3390/foods9030278.
Foodborne pathogens can cause foodborne illness. In reality, one food sample may carry more than one pathogen. A rapid, sensitive, and multiple target method for bacteria detection is crucial in food safety. For the simultaneous detection of , , and Enteritidis, multi-objective recombinase polymerase amplification (RPA) combined with a lateral flow dipstick (LFD) was developed in this study. The whole process, including amplification and reading, can be completed in 15 min at 37 °C. The detection limits were 2.6 × 10 CFU/mL for , 7.6 × 10 CFU/mL for , and 1.29 × 10 CFU/mL for Enteritidis. Moreover, colored signal intensities on test lines were measured by a test strip reader to achieve quantitative detection for (R = 0.9903), (R = 0.9928), and Enteritidis (R = 0.9945). In addition, the method demonstrated good recoveries (92.00%-107.95%) in the testing of spiked food samples. Therefore, the multiplex LFD-RPA assay is a feasible method for the rapid, sensitive, and quantitative detection of bacterial pathogens in seafood.
食源性病原体可导致食源性疾病。实际上，一份食品样本可能携带不止一种病原体。一种快速、灵敏且能检测多种目标的细菌检测方法对食品安全至关重要。在本研究中，开发了一种将多目标重组酶聚合酶扩增（RPA）与侧向流动试纸条（LFD）相结合的方法，用于同时检测[具体细菌名称缺失]、[具体细菌名称缺失]和肠炎沙门氏菌。整个过程，包括扩增和读数，可在37℃下15分钟内完成。[具体细菌名称缺失]的检测限为2.6×10 CFU/mL，[具体细菌名称缺失]为7.6×10 CFU/mL，肠炎沙门氏菌为1.29×10 CFU/mL。此外，通过试纸条读数仪测量测试线上的有色信号强度，以实现对[具体细菌名称缺失]（R = 0.9903）、[具体细菌名称缺失]（R = 0.9928）和肠炎沙门氏菌（R = 0.9945）的定量检测。另外，该方法在加标食品样本检测中显示出良好的回收率（92.00%-107.95%）。因此，多重LFD-RPA检测法是一种用于快速、灵敏且定量检测海鲜中细菌病原体的可行方法。
