This study aims to establish an improved solid phase DNA amplification technique for the detection of DNA biomarkers in the point of care (POC) based molecular diagnostic fields of human diseases including cancer. To date, a complex of DNA biomarkers is known to be associated with human diseases. In particular, many known biomarkers are associated with human cancers, and these need to be simultaneously tested for early detection and prognosis of cancer. In this study, we developed a DNA amplification technique with signal enhancement that combines with solid phase bridge PCR and fluorescence resonance energy transfer (FRET), which can be improved the multiple targets detection simultaneously without obtaining false-positives that caused by the primers binding to each other when multiple targets are amplifying on a solid surface. Here, we showed that the fluorescence signals are 2.5 times enhanced on the Au surface with the KRAS gene (target DNA), which is a common DNA biomarker of human cancers, compared to on the Au surface with non-target DNA by using the present technique. Furthermore, we confirmed the storage conditions of the Au sensor with the immobilized primers for the commercialization as a molecular diagnostic device in the field of POC (point of care)-based cancer applications. Therefore, the present technique will be useful not only for the detection of multiple targets simultaneously without any false-positive signal generated by primers, but also enable integration with a microfluidic system in order to minimize the platform for the genetic analysis of DNA biomarkers in the field of diagnostic applications for disease treatment.
Bibliographical noteFunding Information:
This work was supported by the Agency for Science, Technology and Research (A*STAR) Joint Council Office (JCO) Development Programme Grant ( 1234e00018 ), Singapore.
All Science Journal Classification (ASJC) codes
- Electronic, Optical and Magnetic Materials
- Condensed Matter Physics
- Surfaces, Coatings and Films
- Metals and Alloys
- Electrical and Electronic Engineering
- Materials Chemistry