Abstract
.The focus of this work is toward the development of a point-of-care (POC) handheld technology for the noninvasive early detection of salivary biomarkers. The initial of focus was the detection and quantification of S100 calcium-binding protein P (S100P) mRNA found in whole saliva for use as a potential biomarker for oral cancer. Specifically, a surface-enhanced Raman spectroscopy (SERS)-based approach and assay were designed, developed, and tested for sensitive and rapid detection of S100P mRNA. Gold nanoparticles (AuNPs) were conjugated with oligonucleotides and malachite green isothiocyanate was then used as a Raman reporter molecule. The hybridization of S100P target to DNA-conjugated AuNPs in sandwich assay format in both free solution and a vertical flow chip (VFC) was confirmed using a handheld SERS system. The detection limit of the SERS-based assay in free solution was determined to be 1.1 nM, whereas on the VFC the detection limit was observed to be 10 nM. SERS-based VFCs were also used to quantify the S100P mRNA from saliva samples of oral cancer patients and a healthy group. The result indicated that the amount of S100P mRNA detected for the oral cancer patients is three times higher than that of a healthy group.
Highlights
Oral cancer, of which more than 90% is oral squamous cell carcinoma (OSCC), is the world’s sixth most common cancer.[1]
Its fluorescence intensity at 662 nm is located at approximately half of the intensity of the 130:1 Cy5 DNA oligomer standard solution, which means that only about half of DNA oligomers were conjugated on the AuNPs under the current incubation conditions
SERS bands assigned to Malachite green isothiocynate (MGITC) were not observed for the supernatant, which means most of MGITC is immobilized on the left DNA-conjugated AuNPs
Summary
Of which more than 90% is oral squamous cell carcinoma (OSCC), is the world’s sixth most common cancer.[1] According to the World Health Organization, it has been estimated that each year the number of new cases of oral cancer is 529,000 with more than 300,000 deaths.[2] OSCC survival rates are strongly dependent on the stage at diagnosis. When the disease is at stages III and IV, OSCC can be metastasized and the 5year survival rate for the patients remains at ∼39% to 64% despite an aggressive therapy of chemotherapeutic agents with radiation.[3] early diagnosis of oral cancer is important to improve the therapy.[4] The most common way to diagnose OSCC is through regular check-up by a dentist; if something is detected, oral tissue biopsy is performed followed by a lab test. Systematic review and meta-analysis have revealed that clinical examination alone may not be sufficient for the clinician to perform a biopsy or refer for biopsy for early detection of OSCC.[5,6]
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