Detection of Genetically Modified Rice by Loop-Mediated Isothermal Amplification Assays on a Self-Priming Compartmentali
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Detection of Genetically Modified Rice by Loop-Mediated Isothermal Amplification Assays on a Self-Priming Compartmentalization Chip Guotao Ding 1 & Zengjun Jin 2 & Yunzhe Zhang 3 & Yonghong Han 1 & Guiying Li 2
&
Yongfa Jing 4 & Weihao Li 1
Received: 7 January 2020 / Accepted: 3 May 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract We developed a self-priming compartmentalization (SPC) micro-device made of polymethyl methacrylate (PMMA) and integrated a loop-mediated isothermal amplification (LAMP) system for performing multiplex visual detection. The approach had a high throughput of identification of selectable marker gene (SMG) (phosphomannose isomerase (PMI) gene, hygromycin B phosphotransferase (HPT) gene, noglycoside phosphotransferase II (NPTII), β-glucuronidase (GUS), and CP4-5enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS)) and was able to perform five SMG analyses simultaneously within 1 h. This micro-device can detect five SMGs in one injection, each gene is repeated three times, and each micro-reactor arrays without cross-contamination. The method of extracting genomic DNA from GM rice grains can be very simple and the detection method only requires cell homogenization. In addition, the limit of detections of the method for PMI, HPT, and NPTII were 10−4. The limit of detections of the method for GUS and CP4-EPSPS were 10−5. The results demonstrated that our method could specifically recognize in genetically modified crops. Keywords Self-priming compartmentalization (SPC) micro-device . Loop-mediated isothermal amplification (LAMP) . Genetically modified (GM) . Selectable marker gene (SMG)
Introduction According to the International Service for the Acquisition of Agri-biotech Applications (ISAAA), since the commercialization of GM crops began in the end of the twentieth century, the cumulative planting area of GM crops worldwide has reached 2 billion hectares. In 2017, GM crops were planted in 189.8 million hectares, which the area planted with GM crops increased 3% from 2016, in 24 countries around the Guotao Ding and Zengjun Jin contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12161-020-01766-8) contains supplementary material, which is available to authorized users. * Weihao Li [email protected] 1
Handan Municipal Centre for Disease Control and Prevention, Handan 056008, Hebei Province, China
2
School of Medicine, Hebei University of Engineering, Handan 056002, Hebei Province, China
3
College of Food Science and Technology, Agricultural University of Hebei, Baoding 071001, Hebei Province, China
4
Handan First Hospital, Handan 056000, Hebei Province, China
world (James 2017). The GM crops are continuously expanding; the detection of GM crops is receiving more attention. A large number of researchers have established methods for detecting GM crops. Polymerase chain reaction (PCR) is the most powerful analytical method for nucleic acid detection owing to its high sp
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