Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification

Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modifie...

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Main Authors:	Chen, Xiaoyun, Wang, Xiaofu, Jin, Nuo, Zhou, Yu, Huang, Sainan, Miao, Qingmei, Zhu, Qing, Xu, Junfeng
Format:	Online
Language:	English
Published:	Molecular Diversity Preservation International (MDPI) 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3509588/

id	pubmed-3509588
recordtype	oai_dc
spelling	pubmed-35095882013-01-09 Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification Chen, Xiaoyun Wang, Xiaofu Jin, Nuo Zhou, Yu Huang, Sainan Miao, Qingmei Zhu, Qing Xu, Junfeng Article Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%–0.005% GM), was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB) facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops. Molecular Diversity Preservation International (MDPI) 2012-11-07 /pmc/articles/PMC3509588/ /pubmed/23203072 http://dx.doi.org/10.3390/ijms131114421 Text en © 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0).
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Chen, Xiaoyun Wang, Xiaofu Jin, Nuo Zhou, Yu Huang, Sainan Miao, Qingmei Zhu, Qing Xu, Junfeng
spellingShingle	Chen, Xiaoyun Wang, Xiaofu Jin, Nuo Zhou, Yu Huang, Sainan Miao, Qingmei Zhu, Qing Xu, Junfeng Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
author_facet	Chen, Xiaoyun Wang, Xiaofu Jin, Nuo Zhou, Yu Huang, Sainan Miao, Qingmei Zhu, Qing Xu, Junfeng
author_sort	Chen, Xiaoyun
title	Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
title_short	Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
title_full	Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
title_fullStr	Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
title_full_unstemmed	Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
title_sort	endpoint visual detection of three genetically modified rice events by loop-mediated isothermal amplification
description	Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%–0.005% GM), was 10- to 100-fold greater than that of conventional PCR. Additionally, the LAMP assay coupled with an indicator (SYBR green or HNB) facilitated analysis. These findings revealed that the rapid detection method was suitable as a simple field-based test to determine the status of GM crops.
publisher	Molecular Diversity Preservation International (MDPI)
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3509588/
_version_	1611936435119587328

Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification

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