Magnetically Localized Detection of Amplified DNA Using Biotinylated and Fluorescent Primers and Magnetic Nanoparticles.

Abstract

Quantitative nucleic acid detection is widely used in molecular diagnostics for infectious diseases. Here, we demonstrate that the previously developed MLFIA (magnetically localized fluorescent immunoassay) has the potential to detect Polymerase Chain Reaction (PCR) and loop-mediated isothermal amplification (LAMP) products using biotinylated and fluorescent primers and streptavidin-coated magnetic nanoparticles. The functionalized nanoparticles separate amplified DNA from non-incorporated primers in situ, allowing the quantification of DNA products. We compare magnetically localized fluorescence detection to commercial technologies based on the DNA intercalation of fluorescent dyes. Our system allows the detection of PCR and LAMP products but is approximately 10 times less sensitive than standard commercial assays. Future optimizations, such as enhancing the signal-to-noise ratio and improving nanoparticle functionalization, could significantly increase sensitivity and bring it closer to current diagnostic standards. This work highlights the potential of magnetically localized fluorescence detection to detect DNA.