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Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification
Institute of Agriculture Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China
College of Chemistry and Life Science, Shenyang Normal University, Shenyang 110034, China
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 13 September 2012; in revised form: 8 October 2012 / Accepted: 31 October 2012 / Published: 7 November 2012
Abstract: 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.
Keywords: GMO detection; LAMP; transgenic rice; TT51-1; KMD1; KF6
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MDPI and ACS Style
Chen, X.; Wang, X.; Jin, N.; Zhou, Y.; Huang, S.; Miao, Q.; Zhu, Q.; Xu, J. Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification. Int. J. Mol. Sci. 2012, 13, 14421-14433.
Chen X, Wang X, Jin N, Zhou Y, Huang S, Miao Q, Zhu Q, Xu J. Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification. International Journal of Molecular Sciences. 2012; 13(11):14421-14433.
Chen, Xiaoyun; Wang, Xiaofu; Jin, Nuo; Zhou, Yu; Huang, Sainan; Miao, Qingmei; Zhu, Qing; Xu, Junfeng. 2012. "Endpoint Visual Detection of Three Genetically Modified Rice Events by Loop-Mediated Isothermal Amplification." Int. J. Mol. Sci. 13, no. 11: 14421-14433.