Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food
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DOI：
10.3390/foods9010027
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发表时间：
2020-01-01
期刊：
影响因子：
5.2
通讯作者：
Zhang, Mingzhou
中科院分区：
文献类型：
--
作者：
Li, Jiali;Ma, Biao;Zhang, Mingzhou
沙门氏菌可导致严重的食源性疾病。我们建立了一种检测食品中沙门氏菌的侧流免疫分析结合重组酶聚合酶扩增（LFD-RPA）方法。选择保守片段（fimY）作为目的基因。在最佳条件下（37 ℃，10分钟），在纯培养物中的灵敏度为12个菌落形成单位（CFU）/mL。以16种非沙门氏菌菌株作为对照的测试显示，LFD-RPA对沙门氏菌的fimY基因具有特异性。建立的检测方法可以检测浓度低至1.29 × 10（2）CFU/mL的沙门氏菌在人工污染的样品。该检测水平略高于纯细菌培养物。结合试纸条阅读器，LFD-RPA是一种基于测试线强度（反射光测试线与对照线的比例）定量检测沙门氏菌的可行方法。该方法可在资源有限的地区作为一种潜在的床旁检测方法，为食品安全的诊断提供了新的方法和技术支持。
Salmonella can cause serious foodborne diseases. We have developed a lateral flow immunoassay combined with recombinase polymerase amplification (LFD-RPA) for detection of Salmonella in food. The conserved fragment (fimY) was selected as the target gene. Under an optimal condition (37 degrees C, 10 min), the sensitivity was 12 colony-forming units (CFU)/mL in a pure culture. Testing with 16 non-Salmonella strains as controls revealed that LFD-RPA was specific to the fimY gene of Salmonella. The established assay could detect Salmonella at concentrations as low as 1.29 x 10(2) CFU/mL in artificially contaminated samples. This detection was at a slightly higher level than that for a pure bacterial culture. Combined with the test strip reader, the LFD-RPA is a feasible method for quantitative detection of Salmonella based on the test line intensity, which was the ratio for the test line and control line of the reflected light. The method could be a potential point-of-care test in limited resource areas and provides a new approach and technical support for the diagnosis of food safety.