Increasing the Sensitivity of Aspergillus Galactomannan ELISA Using Silver Nanoparticle-Based Surface-Enhanced Raman Spectroscopy

Galactomannan (GM) is a polysaccharide secreted by opportunistic pathogenic fungi of the Aspergillus genus. It is prescribed as a diagnostic biomarker of invasive aspergillosis in immunocompromised patients by the guidelines for diagnosis and management of Aspergillus diseases. It has been shown previously that the measurement of soluble horseradish peroxidase (HRP) using surface-enhanced Raman scattering (SERS) of 2,3-diaminophenazine enzymatic reaction product on silver nanoparticles is largely superior in detection limit compared to colorimetric readout. In this study, a highly sensitive SERS-based HRP measurement protocol was applied to enzyme-linked immunosorbent assay (ELISA) for GM quantification in biological fluids. The detection limit for GM was 4.3 pg per sample, which is one and a half orders of magnitude lower compared to colorimetric detection with o-phenylenediamine as a substrate and five times more sensitive than ELISA using 3,3',5,5'-tetramethylbenzidine.