Development of surface-enhanced Raman scattering nanoprobes for the simultaneous multiplex detection of...

Relying on a single biomarker in biomedical analysis is often insufficient for accurate disease or pathogen determination. A recent trend is using simultaneous multiplex detection of multiple biomarkers to improve diagnostic accuracy and throughput. To enable multiplex detection, we developed a series of surface-enhanced Raman scattering (SERS) nanoprobes, referred to as nanoaggregate-embedded beads (NAEBs). These NAEBs were synthesized using three distinct Raman reporter molecules: Safranin O, ethyl violet, and cresyl violet acetate. By integrating the NAEBs with magnetic nanoparticles and a simple capillary magnetofluidic device, we developed a rapid and simultaneous multiplex detection platform for genetic analysis of an aquacultural pathogen Vibrio parahaemolyticus (VP) for pirA, pirB, and ompA and genotyping of Alzheimer's disease's risk factor biomarker Apoliproprotein E (ApoE). For VP detection, a limit of detection (LOD) as low as ~ 10^-16 M and a broad dynamic range of six orders were found . For ApoE genotyping, the platform was capable of detecting single nucleotide polymorphisms (SNPs) of rs429358 and rs7412 in the ApoE gene with a LOD of approximately 10^-13 M. These results highlight the potential of this SERS-based simultaneous multiplex detection platform for sensitive and quantitative determination of multiple DNA biomarkers as well as SNPs in complex biological matrices.