Development of a free-solution SERS-based assay for point-of-care oral cancer biomarker detection using DNA-conjugated gold nanoparticles

Sungyub Han; Andrea K. Locke; Luke A. Oaks; Yi-Shing Lisa Cheng; Gerard L. Coté

doi:10.1117/12.2290516

20 February 2018 Development of a free-solution SERS-based assay for point-of-care oral cancer biomarker detection using DNA-conjugated gold nanoparticles

Sungyub Han, Andrea K. Locke, Luke A. Oaks, Yi-Shing Lisa Cheng, Gerard L. Coté

Author Affiliations +

Proceedings Volume 10501, Optical Diagnostics and Sensing XVIII: Toward Point-of-Care Diagnostics; 1050104 (2018) https://doi.org/10.1117/12.2290516
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

It is estimated that the number of new cases of oral cancers worldwide is 529,000 and more than 300,000 deaths each year. The five-year survival rate remains about 50%, and the low survival rate is believed to be due to delayed detection. The primary detection method is through a comprehensive clinical examination by a dentist followed by a biopsy of suspicious lesions. Systematic review and meta-analysis have revealed that clinical examination alone may not be sufficient to cause the clinician to perform a biopsy or refer for biopsy for early detection of OSCC. Therefore, a non-invasive, point-of-Care (POC) detection with high sensitivity and specificity for early detection would be urgently needed, and using salivary biomarkers would be an ideal technology for it. S100 calcium binding protein P (S100P) mRNA presenting in saliva is a potential biomarker for detection of oral cancer. Further, surface enhanced Raman spectroscopy (SERS) has been shown to be a promising POC diagnostic technique. In this research, a SERS-based assay using oligonucleotide strains was developed for the sensitive and rapid detection of S100P. Gold nanoparticles (AuNPs) as a SERS substrate were used for the conjugation with one of two unique 24 base pair oligonucleotides, referred to as left and right DNA probes. A Raman reporter molecule, malachite green isothiocyanate (MGITC), was bound to left-probe-conjugated AuNPs. UV-vis spectroscopy was employed to monitor the conjugation of DNA probes to AuNPs. The hybridization of S100P target to DNA-conjugated AuNPs in sandwich-assay format was confirmed by Raman spectroscopy and shown to yield and R² of 0.917 across the range of 0-200 nM and a limit of detection of 3 nM.

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Sungyub Han, Andrea K. Locke, Luke A. Oaks, Yi-Shing Lisa Cheng, and Gerard L. Coté "Development of a free-solution SERS-based assay for point-of-care oral cancer biomarker detection using DNA-conjugated gold nanoparticles", Proc. SPIE 10501, Optical Diagnostics and Sensing XVIII: Toward Point-of-Care Diagnostics, 1050104 (20 February 2018); https://doi.org/10.1117/12.2290516

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