Journal
ACS APPLIED MATERIALS & INTERFACES
Volume 14, Issue 24, Pages 27666-27674Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsami.2c06061
Keywords
nanoPCR; gold nanoparticles; LAMP; biosensors; proteins; GSH
Funding
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- National Natural Science Foundation of China [21575165]
- China Scholarship Council
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This study systematically investigated the effect of surface-modified gold nanoparticles (AuNPs) on loop-mediated isothermal amplification (LAMP). It was found that mixing AuNPs with primers delayed the reaction, while other mixing orders accelerated it. The performance of AuNPs on LAMP was most significant when they formed a protein-like surface. By using glutathione-capped AuNPs, a detection limit of around 100 copies/μL of target DNA was achieved, indicating a higher sensitivity in DNA sensing.
Loop-mediated isothermal amplification (LAMP) is a popular DNA amplification method. Gold nanoparticles (AuNPs) were reported to enhance the efficiency of LAMP, although the underlying mechanism remained elusive. Since AuNPs strongly adsorb a range of ligands, preadsorbed ligands cannot be easily displaced. In this work, we systematically investigated the effect of surface-modified AuNPs on LAMP by varying the order of mixing of AuNPs with each reagent in the LAMP system (Mg2+, template DNA, dNTPs, primers, and polymerase). Mixing the AuNPs with the primers delayed the LAMP based on SYBR green I fluorescence. While other orders of mixing had little effect, all accelerated the reaction. We then tested other common ligands including polymers (polyethylene glycol and polyvinylpyrrolidone), inorganic ions (Br-), proteins, glutathione (GSH), and DNA (A(15)) on AuNP-LAMP. The boosted AuNP performance on LAMP was most obvious when the AuNPs formed a protein-like surface. Finally, using GSH-capped AuNPs, a detection limit of around 100 copies/mu L-1 of target DNA was achieved. This work has identified a ligand-capped AuNP strategy to boost LAMP and yielded a higher sensitivity in DNA sensing, which also deepens our understanding of AuNP-assisted LAMP.
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