Detection of C-reactive protein using single cluster analysis of gold nanoparticle aggregates using a dark-field microscope equipped with a smartphone†

Gold nanoparticles (AuNPs), which have been used as colorimetric biosensors, show strong light scattering, allowing individual AuNPs to be identified using a dark-field microscope (DFM). In this study, we developed a method of observing the target molecule-derived aggregation of AuNPs modified with DNA aptamers at the single-cluster level using the DFM. C-Reactive protein (CRP) is an important clinical biomarker of inflammatory and cardiovascular diseases, for which a simple, inexpensive, and sensitive detection method is needed. In this study, the CRP-mediated aggregate formation of CRP aptamer-modified AuNPs was evaluated with single-cluster analysis using the DFM, and the detection limit was 17 nM, which was sufficient as a diagnostic indicator for CRP. We also developed a portable DFM equipped with a smartphone and a stage adjustment system, which enables single-cluster observation of AuNPs, and showed that 50 nM of CRP could be detected, indicating that this approach is suitable for point-of-care diagnosis. With the selection of appropriate aptamers, this method can be applied for the detection of various molecules.