Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification

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...

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Bibliographic Details
Published in:Food Chemistry: Molecular Sciences
Main Authors: Li, Qiuping, Xue, Hanyue, Fei, Yanjin, Cao, Min, Xiong, Xiaohui, Xiong, Xiong, Yang, Ying, Wang, Libin
Format: Text
Language:English
Published: Elsevier 2022
Subjects:
Research Article
Atlantic salmon
Salmo salar
Online Access:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9235052/
http://www.ncbi.nlm.nih.gov/pubmed/35769395
https://doi.org/10.1016/j.fochms.2022.100107
Description
Summary: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 fluorophores (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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