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Identification of pork in meat products using real-time loop-mediated isothermal amplification

In this study, a one-step, real-time, loop-mediated isothermal amplification (RealAmp) assay was developed, for the highly specific detection of pork DNA. For the assay, the mtDNA of cytochrome b (cytb) gene was amplified at 63 °C using SYBR Green I for 45 min with a Real-Time Polymerase Chain React...

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Detalles Bibliográficos
Autores principales: Yang, Lixia, Fu, Shujun, Peng, Xinkai, Li, Le, Song, Taoping, Li, Lin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4433938/
https://www.ncbi.nlm.nih.gov/pubmed/26019573
http://dx.doi.org/10.1080/13102818.2014.963789
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author Yang, Lixia
Fu, Shujun
Peng, Xinkai
Li, Le
Song, Taoping
Li, Lin
author_facet Yang, Lixia
Fu, Shujun
Peng, Xinkai
Li, Le
Song, Taoping
Li, Lin
author_sort Yang, Lixia
collection PubMed
description In this study, a one-step, real-time, loop-mediated isothermal amplification (RealAmp) assay was developed, for the highly specific detection of pork DNA. For the assay, the mtDNA of cytochrome b (cytb) gene was amplified at 63 °C using SYBR Green I for 45 min with a Real-Time Polymerase Chain Reaction (PCR) System that measured the fluorescent signal at one-minute intervals. As little as 1 pg of template DNA could be detected, without any cross-reactivity with non-target species. Meat mixtures, heat-treated at 100 °C for 15 min, prepared by mixing pork meat with beef at different ratios (0.01%–10%) were tested, and the RealAmp assays allowed the detection of as little as 0.01% pork in the meat mixtures. Thus, this work showed that RealAmp could be used for specific identification and sensitive quantification of meat species, even for heat-treated meat products.
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spelling pubmed-44339382015-05-25 Identification of pork in meat products using real-time loop-mediated isothermal amplification Yang, Lixia Fu, Shujun Peng, Xinkai Li, Le Song, Taoping Li, Lin Biotechnol Biotechnol Equip Article; Food Biotechnology In this study, a one-step, real-time, loop-mediated isothermal amplification (RealAmp) assay was developed, for the highly specific detection of pork DNA. For the assay, the mtDNA of cytochrome b (cytb) gene was amplified at 63 °C using SYBR Green I for 45 min with a Real-Time Polymerase Chain Reaction (PCR) System that measured the fluorescent signal at one-minute intervals. As little as 1 pg of template DNA could be detected, without any cross-reactivity with non-target species. Meat mixtures, heat-treated at 100 °C for 15 min, prepared by mixing pork meat with beef at different ratios (0.01%–10%) were tested, and the RealAmp assays allowed the detection of as little as 0.01% pork in the meat mixtures. Thus, this work showed that RealAmp could be used for specific identification and sensitive quantification of meat species, even for heat-treated meat products. Taylor & Francis 2014-09-03 2014-10-22 /pmc/articles/PMC4433938/ /pubmed/26019573 http://dx.doi.org/10.1080/13102818.2014.963789 Text en © 2014 The Author(s). Published by Taylor & Francis. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0/, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.
spellingShingle Article; Food Biotechnology
Yang, Lixia
Fu, Shujun
Peng, Xinkai
Li, Le
Song, Taoping
Li, Lin
Identification of pork in meat products using real-time loop-mediated isothermal amplification
title Identification of pork in meat products using real-time loop-mediated isothermal amplification
title_full Identification of pork in meat products using real-time loop-mediated isothermal amplification
title_fullStr Identification of pork in meat products using real-time loop-mediated isothermal amplification
title_full_unstemmed Identification of pork in meat products using real-time loop-mediated isothermal amplification
title_short Identification of pork in meat products using real-time loop-mediated isothermal amplification
title_sort identification of pork in meat products using real-time loop-mediated isothermal amplification
topic Article; Food Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4433938/
https://www.ncbi.nlm.nih.gov/pubmed/26019573
http://dx.doi.org/10.1080/13102818.2014.963789
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