Development of Detection System Using Multiplex PCR and Liquid Beadarray for Stacked Genetically Modified Rice Event (LS28×Cry1Ac)

• Published in: Applied biological chemistry Vol. 53; no. 5; pp. 639 - 646
• Main Authors: Choi, S.H., Seoul Women's University, Seoul, Republic of Korea, Oh, Y.T., YeBT Co., Ltd., Seoul, Republic of Korea, Kwon, J.Y., Seoul Women's University, Seoul, Republic of Korea, Lee, S.N., Seoul Women's University, Seoul, Republic of Korea, Han, B.D., YeBT Co., Ltd., Seoul, Republic of Korea, Ryu, K.H., Seoul Women's University, Seoul, Republic of Korea
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01.10.2010; Springer Nature B.V; 한국응용생명화학회
• Subjects: Applied Microbiology; Biological Techniques; Bioorganic Chemistry; Biotinylation; Chemistry; Chemistry and Materials Science; Cry1Ac toxin; detection; Fluorescence; Food Science/Microbiology; Genetic modification; Hybridization; liquid beadarray; multiplex PCR; Multiplexing; PCR; Polymerase chain reaction; Primers; Qualitative analysis; Rice; SBS primer; Sequences; stacked GM crop; Target detection; Transgenes; 농학; multiplex PCR; liquid beadarray; detection; stacked GM crop; SBS primer
• ISSN: 1738-2203; 2468-0834; 2234-344X; 2468-0842
• DOI: 10.3839/jksabc.2010.097

A multiplex system was developed to assess detection of stacked genetically modified (GM) rice (LS28 × Cry1Ac) based on multiplex polymerase chain reaction (PCR) and liquid beadarray, and the accuracy of the system was analyzed. Standard and specific bulging specific (SBS) primers with standard primers were used to simultaneously detect multiple targets in stacked events of rice. Five sets of primers for the stacked events were applied to amplify their targets, and were separated distinctly in agarose gel. A liquid beadarray assay for the stacked GM rice was performed using the multiplex PCR products, followed by target biotinylation and hybridization between biotinylated-tagged target and anti-tagged bead. Fluorescent signals of the hybridized target sequences were detected by the Luminex system. The signaling patterns were analyzed by their mean fluorescent intensity (MFI) value. Results showed that liquid beadarrays with standard and SBS primers were in complete agreement with the PCR data, and detection of the different target elements was found to be very specific with no cross reaction among samples. Therefore, our detection system developed for stacked GM crop using multiplex PCR and liquid beadarray can be a useful and efficient system for screening and analyzing multiple transgenes in a single tube for qualitative analysis.