Detection of Genetically Modified Rice by Loop-Mediated Isothermal Amplification Assays on a Self-Priming Compartmentalization Chip

Autor: Weihao Li, Yongfa Jing, Guotao Ding, Zengjun Jin, Yunzhe Zhang, Guiying Li, Yonghong Han
Rok vydání: 2020
Předmět:
Zdroj: Food Analytical Methods. 13:1454-1461
ISSN: 1936-976X
1936-9751
DOI: 10.1007/s12161-020-01766-8
Popis: 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 CP4-5-enolpyruvylshikimate-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.
Databáze: OpenAIRE
Externí odkaz: