期刊
FOOD CHEMISTRY: MOLECULAR SCIENCES
卷 4, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.fochms.2022.100107
关键词
Loop-mediated isothermal amplification (LAMP); Oncorhynchus mykiss; Salmo salar; Species identification; Visual detection
资金
- Natural Science Research Program of Colleges and Universities in Jiangsu Province [21KJB550011]
- National Natural Science Foundation of China [31701688]
This study achieved specific detection in LAMP using fluorescently labeled probes, with the DNA produced being precipitated and appearing as different colors for different target species. The optimized duplex LAMP system can visually detect at least 1 ng of DNA.
Loop-mediated isothermal amplification (LAMP) is often confounded by the non-specific amplification arising from primer dimers, off-target priming, and other artifacts. Precipitation of the DNA produced during LAMP with the use of specific fluorescently labeled probe has proved the effectiveness in specific detection. Herein, two fluomphores (ROX and FAM) were attached to the primers S-LB-6 and R-FIP for Atlantic salmon and rainbow trout, respectively, which are self-quenched in unbound state and become de-quenched after binding to the dumbbell-shaped DNA specifically. The DNA precipitation and appearance of small sediment took 10 s of centrifugation at 1000 g, by adding polyethylenimine (PEI) 600. Each target species was specifically amplified with the predicted color of PEI-DNA sediment, namely red for Atlantic salmon, green for rainbow trout, and pale yellow for mixed species. The optimized duplex LAMP system has proved its specificity and can detect as little as 1 ng DNA in visual detection.
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