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Potential of cross-priming amplification and DNA-based lateral-flow strip biosensor for rapid on-site GMO screening
Authors:Xin Huang  Congcong Zhai  Qimin You  Hongjun Chen
Institution:1. Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China
2. Ustar Biotechnologies (Hangzhou) Ltd, Hangzhou, 310053, China
Abstract:The requirement to monitor the presence of genetically modified organisms (GMO) in a variety of marked products has generated an increasing demand for reliable, rapid, and time and cost-effective analytical methods. Here we report an on-site method for rapid detection of cauliflower mosaic virus promoter (CaMV 35S), a common element present in most GMO, using cross-priming amplification (CPA) technology. Detection was achieved using a DNA-based contamination-proof strip biosensor. The limit of detection was 30 copies for the pBI121 plasmid containing the CaMV 35S gene. The certified reference sample of GM maize line MON810 was detectable even at the low relative mass concentration of 0.05 %. The developed CPA method had high specificity for the CaMV 35S gene, as compared with other GM lines not containing this gene and non-GM products. The method was further validated using nine real-world samples, and the results were confirmed by real-time PCR analysis. Because of its simplicity, rapidity, and high sensitivity, this method of detecting the CaMV 35S gene has great commercial prospects for rapid GMO screening of high-consumption food and agriculture products.
Keywords:
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