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Nanoparticle-mediated light-driven LAMP combined with test strips for sensitive and rapid visual detection of
Yu-E Gao1, Qing Huang2, Yue Huang1
1State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, China; International Joint Laboratory on Food Safety, Institute of Analytical Food Safety, School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.
Abstract:
Antibiotic resistance genes (ARGs) are markers of drug-resistant pathogens, monitoring them contributes to prevent resistance to drugs. The detection methods for ARGs including PCR and isothermal amplification are sensitive and selective. However, it may take several hours or cannot be used on spot. Here, a detection method was developed based on a novel nanoparticle-mediated light-driven (LD) loop-mediated isothermal amplification (LAMP) combining with test strips, and a commonly found methicillin-resistant gene mecA was analyzed to verify the method. Under laser irradiation, gold nanoparticles produced localized surface plasmon resonance. Therefore, they provided both light-induced electrons and localized heat, the former acted as the catalyst of LD-LAMP and the latter as the energy supply. The mecA was amplified, producing numerous double-labeled amplicons. The LD-LAMP was three times as efficient as metal-bathed LAMP. A visual test strip (TS) was designed based on sandwich lateral flow chromatography, reading a LAMP result within 5-10 min. The method has a detection limit of 13.8 copies/μL, with a linear range of 13.8 copies/μL∼1.38 × 107 copies/μL. The LD-LAMPTS was applied to the on-site detection of mecA in milk samples. The results were consistent with qPCR, and total detection time reduced from 1 h by qPCR to 20-25 min by LD-LAMPTS.
