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Application of Recombinase Polymerase Amplification with Lateral Flow for a Naked-Eye Detection of <i>Listeria monocytogenes</i> on Food Processing Surfaces
oleh: Sarah Azinheiro, Joana Carvalho, Marta Prado, Alejandro Garrido-Maestu
Format: | Article |
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Diterbitkan: | MDPI AG 2020-09-01 |
Deskripsi
The continuous contamination of foods with <i>L. monocytogenes</i>, highlights the need for additional controls in the food industry. The verification of food processing plants is key to avoid cross-contaminations, and to assure the safety of the food products. In this study, a new methodology for the detection of <i>L. monocytogenes</i> on food contact surfaces was developed and evaluated. It combines Recombinase Polymerase Amplification (RPA) with the lateral flow (LF) naked-eye detection. Different approaches for the recovery of the bacteria from the surface, the enrichment step and downstream analysis by RPA-LF were tested and optimized. The results were compared with a standard culture-based technique and qPCR analysis. Sampling procedure with sponges was more efficient for the recovery of the bacteria than a regular swab. A 24 h enrichment in ONE broth was needed for the most sensitive detection of the pathogen. By RPA-LF, it was possible to detect 1.1 pg/µL of pure <i>L. monocytogenes</i> DNA, and the complete methodology reached a LoD<sub>50</sub> of 4.2 CFU/cm<sup>2</sup> and LoD<sub>95</sub> of 18.2 CFU/cm<sup>2</sup>. These results are comparable with the culture-based methodology and qPCR. The developed approach allows for a next-day detection without complex equipment and a naked-eye visualization of the results.