Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow Immunoassay for Rapid Detection of Salmonella in Food

被引:63
作者
Li, Jiali [1 ]
Ma, Biao [1 ]
Fang, Jiehong [1 ]
Zhi, Antong [1 ]
Chen, Erjing [1 ]
Xu, Ying [1 ]
Yu, Xiaoping [1 ]
Sun, Chuanxin [2 ]
Zhang, Mingzhou [1 ]
机构
[1] China Jiliang Univ, Zhejiang Prov Key Lab Biometrol & Inspect & Quara, Hangzhou 310018, Peoples R China
[2] Swedish Univ Agr Sci SLU, Linnean Ctr Plant Biol, Dept Plant Biol, Uppsala BioCtr, POB 7080, SE-75007 Uppsala, Sweden
关键词
recombinase polymerase amplification; lateral flow dipstick; Salmonella; rapid and quantitative detection; PREVALENCE; DIPSTICK; FIMY;
D O I
10.3390/foods9010027
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
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.
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页数:12
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