RESUMO
High-impact diseases, including cancer, cardiovascular disease, and neurological disease, are challenging to diagnose without supplementing clinical evaluation with laboratory testing. Even with laboratory tools, definitive diagnosis often remains elusive. The lack of three crucial elements presents a road block to achieving the potential of clinical diagnostic tests: (1) definitive disease-associated protein and genetic markers, (2) easy and inexpensive sampling methods with minimal discomfort for the subject, and (3) an accurate and quantitative diagnostic platform. Our aim is to develop and validate a solution for requirement (3) and also to develop a portable system. Requirements (1) and (2) will be addressed through the utilization of novel and highly specific oral cancer saliva proteomic and genomic biomarkers and the use of saliva as the biofluid of choice, respectively. The Oral Fluid NanoSensor Test (OFNASET) technology platform combines cutting-edge technologies, such as self-assembled monolayers (SAM), bionanotechnology, cyclic enzymatic amplification, and microfluidics, with several well-established techniques including microinjection molding, hybridization-based detection, and molecular purification. The intended use of the OFNASET is for the point of care multiplex detection of salivary biomarkers for oral cancer. We have demonstrated that the combination of two salivary proteomic biomarkers (thioredoxin and IL-8) and four salivary mRNA biomarkers (SAT, ODZ, IL-8, and IL-1b) can detect oral cancer with high specificity and sensitivity. Our preliminary studies have shown compelling results. We sequentially delivered a serial dilution of IL-8 antigen, probe solution, wash, enzyme solution, wash, and mediator solution to sensor reaction chambers housed in a prototype cartridge and demonstrated strong signal separation at 50 pg/mL above a negative control.
Assuntos
Técnicas Biossensoriais , Eletroquímica , Nanotecnologia , Saliva/química , Técnicas Biossensoriais/métodos , Eletroquímica/métodos , Humanos , Nanotecnologia/métodosRESUMO
With the growing number of fatalities resulting from the 100 or so cancer-related diseases, new enabling tools are required to provide extensive molecular profiles of patients to guide the clinician in making viable diagnosis and prognosis. Unfortunately with cancer-related diseases, there is not one molecular marker that can provide sufficient information to assist the clinician in making effective prognoses or even diagnoses. Indeed, large panels of markers must typically be evaluated that cut across several different classes (mutations in certain gene fragments--DNA; over/under-expression of gene activity as monitored by messenger RNAs; the amount of proteins present in serum or circulating tumor cells). The classical biosensor format (dipstick approach for monitoring the presence of a single element) is viewed as a valuable tool in many bioassays, but possesses numerous limitations in cancer due primarily to the single element nature of these sensing platforms. As such, if biosensors are to become valuable tools in the arsenal of the clinician to manage cancer patients, new formats are required. This review seeks to provide an overview of the current thinking on molecular profiling for diagnosis and prognosis of cancers and also, provide insight into the current state-of-the-art in the biosensor field and new strategies that must be considered to bring this important technology into the cancer field.