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Rapid Visual LAMP Method for Detection of Genetically Modified Organisms

[Image: see text] We developed a novel loop-mediated isothermal amplification (LAMP) method using DNA captured on polyacrylamide microparticles (PAMMPs) as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection of genetically modified organisms (GMOs). Here, DNA was extracted by a fast and cost...

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Autores principales: Xing, Yujun, Liang, Jie, Dong, Fei, Wu, Jirong, Shi, Jianrong, Xu, Jianhong, Wang, Jinke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2023
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10433496/
https://www.ncbi.nlm.nih.gov/pubmed/37599972
http://dx.doi.org/10.1021/acsomega.3c03567
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author Xing, Yujun
Liang, Jie
Dong, Fei
Wu, Jirong
Shi, Jianrong
Xu, Jianhong
Wang, Jinke
author_facet Xing, Yujun
Liang, Jie
Dong, Fei
Wu, Jirong
Shi, Jianrong
Xu, Jianhong
Wang, Jinke
author_sort Xing, Yujun
collection PubMed
description [Image: see text] We developed a novel loop-mediated isothermal amplification (LAMP) method using DNA captured on polyacrylamide microparticles (PAMMPs) as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection of genetically modified organisms (GMOs). Here, DNA was extracted by a fast and cost-effective method using PAMMPs. Four LAMP primers were designed for the PAMMPs@DNA-LAMP method to detect the cauliflower mosaic virus 35S (CaMV35S) promotor in GMOs. We thus developed this method for rapid extraction of DNA (5–10 min) and fast amplification of DNA within ∼30 min at a constant temperature of 63 °C. Moreover, the DNA captured by PAMMPs (PAMMPs@DNA) could be effectively detected by both conventional and quantitative PCR (qPCR) and LAMP. The PAMMPs@DNA-LAMP method was validated with high specificity, sensitivity, and performance for practical sample analysis. This assay detected 0.01% target sequences, which had a high specificity like qPCR and better than the conventional PCR (cPCR). Furthermore, PAMMPs@DNA-LAMP was successfully used to extract and detect DNA from food samples of the major crops (soybean, maize, rice, etc.). In summary, a novel PAMMPs@DNA-LAMP assay has been developed, which has higher sensitivity and spends less time than the cPCR detection using the conventional DNA extracted process. This method offers a novel approach for rapid detection of GMOs in the field.
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spelling pubmed-104334962023-08-18 Rapid Visual LAMP Method for Detection of Genetically Modified Organisms Xing, Yujun Liang, Jie Dong, Fei Wu, Jirong Shi, Jianrong Xu, Jianhong Wang, Jinke ACS Omega [Image: see text] We developed a novel loop-mediated isothermal amplification (LAMP) method using DNA captured on polyacrylamide microparticles (PAMMPs) as templates (PAMMPs@DNA-LAMP) for rapid qualitative detection of genetically modified organisms (GMOs). Here, DNA was extracted by a fast and cost-effective method using PAMMPs. Four LAMP primers were designed for the PAMMPs@DNA-LAMP method to detect the cauliflower mosaic virus 35S (CaMV35S) promotor in GMOs. We thus developed this method for rapid extraction of DNA (5–10 min) and fast amplification of DNA within ∼30 min at a constant temperature of 63 °C. Moreover, the DNA captured by PAMMPs (PAMMPs@DNA) could be effectively detected by both conventional and quantitative PCR (qPCR) and LAMP. The PAMMPs@DNA-LAMP method was validated with high specificity, sensitivity, and performance for practical sample analysis. This assay detected 0.01% target sequences, which had a high specificity like qPCR and better than the conventional PCR (cPCR). Furthermore, PAMMPs@DNA-LAMP was successfully used to extract and detect DNA from food samples of the major crops (soybean, maize, rice, etc.). In summary, a novel PAMMPs@DNA-LAMP assay has been developed, which has higher sensitivity and spends less time than the cPCR detection using the conventional DNA extracted process. This method offers a novel approach for rapid detection of GMOs in the field. American Chemical Society 2023-08-01 /pmc/articles/PMC10433496/ /pubmed/37599972 http://dx.doi.org/10.1021/acsomega.3c03567 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Xing, Yujun
Liang, Jie
Dong, Fei
Wu, Jirong
Shi, Jianrong
Xu, Jianhong
Wang, Jinke
Rapid Visual LAMP Method for Detection of Genetically Modified Organisms
title Rapid Visual LAMP Method for Detection of Genetically Modified Organisms
title_full Rapid Visual LAMP Method for Detection of Genetically Modified Organisms
title_fullStr Rapid Visual LAMP Method for Detection of Genetically Modified Organisms
title_full_unstemmed Rapid Visual LAMP Method for Detection of Genetically Modified Organisms
title_short Rapid Visual LAMP Method for Detection of Genetically Modified Organisms
title_sort rapid visual lamp method for detection of genetically modified organisms
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10433496/
https://www.ncbi.nlm.nih.gov/pubmed/37599972
http://dx.doi.org/10.1021/acsomega.3c03567
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