Orsini Etienne, Bruckert Franz, Weidenhaupt Marianne, Cugat Orphée, Kauffmann Paul, Delshadi Sarah
MagIA Diagnostics, 15 Rue Maréchal Leclerc, 38130 Échirolles, France.
Univ. Grenoble Alpes, Centre National de la Recherche Scientifique Grenoble Institut National Polytechnique, Laboratoire des Matériaux et du Génie Physique, 38000 Grenoble, France.
Biosensors (Basel). 2025 Mar 18;15(3):195. doi: 10.3390/bios15030195.
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.
定量核酸检测在传染病的分子诊断中被广泛应用。在此，我们证明了先前开发的磁定位荧光免疫分析（MLFIA）有潜力使用生物素化荧光引物和链霉亲和素包被的磁性纳米颗粒来检测聚合酶链反应（PCR）和环介导等温扩增（LAMP）产物。功能化的纳米颗粒在原位将扩增的DNA与未掺入的引物分离，从而实现DNA产物的定量。我们将磁定位荧光检测与基于荧光染料DNA嵌入的商业技术进行了比较。我们的系统能够检测PCR和LAMP产物，但灵敏度比标准商业检测低约10倍。未来的优化，如提高信噪比和改善纳米颗粒功能化，可能会显著提高灵敏度并使其更接近当前的诊断标准。这项工作突出了磁定位荧光检测在检测DNA方面的潜力。
