Abstract
Loop-mediated isothermal amplification (LAMP) had been employed as a powerful tool to facilitate genetic tests for various food pathogens, as it is easy to perform. Recently, various methods of detecting the LAMP amplicon were developed. In this study, we improved two LAMP assays by combining LAMP with chromatographic flow dipstick (LFD) assays for Salmonella (targeting phoP and invA, respectively). We evaluated different labeled primer sets, then selected the optimal sets to perform the LFD assays. We compared the optimal LFD and LAMP assays with the ISO standard method. The results showed that LFD was more sensitive and quicker than LAMP. Furthermore, enrichment broths of 225 food samples were tested. The sensitivity of two LFD assays was 100%. The specificity of LFD assay targeting phoP was 99.1%, and LFD assay targeting invA was 99.5%. For the LFD assay targeting phoP, the estimate of limit of detection (LOD) 50% was 0.061 CFU/g and the estimate of LOD 95% was 0.265 CFU/g. For the LFD assay targeting invA, the estimate of LOD 50% was 0.040 CFU/g and the estimate of LOD 95% was 0.172 CFU/g. We validated this method in a primary laboratory, where we accomplished the assay only using an incubator and a heating block. It suggested that the LFD assay had the potential to become a suitable diagnostic method in field test and primary labs.
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The authors would like to thank Dr. Shuo Yang for her expert review of the manuscript.
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This study was supported by the Fundamental Research Funds for the Central Universities, Nankai University (No. 6391159).
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All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Jia Yu, Jiahua Xing, Xijing Zhan, and Zhen Yang. The first draft of the manuscript was written by Jia Yu, Yin Liu, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Jia Yu declares that she has no conflict of interest. Jiahua Xing declares that he has no conflict of interest. Xijing Zhan declares that she has no conflict of interest. Zhen Yang declares that he has no conflict of interest. Jun Qi declares that he has no conflict of interest. Yuxi Wei declares that he has no conflict of interest. Yin Liu declares that he has no conflict of interest.
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Yu, J., Xing, J., Zhan, X. et al. Improvement of Loop-Mediated Isothermal Amplification Combined with Chromatographic Flow Dipstick Assay for Salmonella in Food Samples. Food Anal. Methods 13, 1398–1408 (2020). https://doi.org/10.1007/s12161-020-01760-0
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DOI: https://doi.org/10.1007/s12161-020-01760-0