期刊
FOOD CHEMISTRY
卷 405, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2022.134975
关键词
Loop-mediated isothermal amplification; (LAMP); Salmo salar; Species identification; Visual detection; Closed-tube
To improve the suitability for naked-eye inspection, a strategy based on the fluorescence property of SYTO 9 and HNB was proposed, and a novel dual-color fluorescence LAMP (dfLAMP) assay was developed for visual detection of Atlantic salmon. The optimized dfLAMP assay has proved its specificity and can detect as little as 1 fg Atlantic salmon DNA.
Loop-mediated isothermal amplification (LAMP) visual detection based on hydroxyl naphthol blue (HNB) and SYTO 9 is often confounded by the narrow color variation window and the requirement of empirical preset of cutoff intensity value. To improve the suitability for naked-eye inspection, the present work proposed a strategy based on the fluorescence property of SYTO 9 and HNB to enlarge the contrast and a novel dual-color fluores-cence LAMP (dfLAMP) assay was developed for visual detection of Atlantic salmon. Specifically, HNB of 26.25 mu M, blended with SYTO 9 of 0.75-1.5 mu M, was added in the mixture before amplification, producing light green fluorescence for both positive and negative samples. After amplification, green or yellow-green fluorescence was observed for positive samples, while only orange-red fluorescence emitted for negative ones, enabling an easy and rapid distinguish. The optimized dfLAMP assay has proved its specificity and can detect as little as 1 fg Atlantic salmon DNA.
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