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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ftpubmed:oai:pubmedcentral.nih.gov:9235052 2023-05-15T15:30:11+02:00 Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification Li, Qiuping Xue, Hanyue Fei, Yanjin Cao, Min Xiong, Xiaohui Xiong, Xiong Yang, Ying Wang, Libin 2022-03-26 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 en eng Elsevier http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9235052/ http://www.ncbi.nlm.nih.gov/pubmed/35769395 http://dx.doi.org/10.1016/j.fochms.2022.100107 © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). CC-BY-NC-ND Food Chem (Oxf) Research Article Text 2022 ftpubmed https://doi.org/10.1016/j.fochms.2022.100107 2022-07-03T00:50:38Z 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. Text Atlantic salmon Salmo salar PubMed Central (PMC) Food Chemistry: Molecular Sciences 4 100107 |
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Research Article Li, Qiuping Xue, Hanyue Fei, Yanjin Cao, Min Xiong, Xiaohui Xiong, Xiong Yang, Ying Wang, Libin Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
topic_facet |
Research Article |
description |
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. |
format |
Text |
author |
Li, Qiuping Xue, Hanyue Fei, Yanjin Cao, Min Xiong, Xiaohui Xiong, Xiong Yang, Ying Wang, Libin |
author_facet |
Li, Qiuping Xue, Hanyue Fei, Yanjin Cao, Min Xiong, Xiaohui Xiong, Xiong Yang, Ying Wang, Libin |
author_sort |
Li, Qiuping |
title |
Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
title_short |
Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
title_full |
Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
title_fullStr |
Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
title_full_unstemmed |
Visual detection of rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
title_sort |
visual detection of rainbow trout (oncorhynchus mykiss) and atlantic salmon (salmo salar) simultaneously by duplex loop-mediated isothermal amplification |
publisher |
Elsevier |
publishDate |
2022 |
url |
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 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
Food Chem (Oxf) |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9235052/ http://www.ncbi.nlm.nih.gov/pubmed/35769395 http://dx.doi.org/10.1016/j.fochms.2022.100107 |
op_rights |
© 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
op_rightsnorm |
CC-BY-NC-ND |
op_doi |
https://doi.org/10.1016/j.fochms.2022.100107 |
container_title |
Food Chemistry: Molecular Sciences |
container_volume |
4 |
container_start_page |
100107 |
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1766360624411967488 |