Development of point-of-care diagnostic tools based on the conformational switch of oligonucleotides – OligoSwitch

We propose to develop «split-quadruplex« oligonucleotide-based sensing platforms for the detection of DNA mutations and protein biomarkers. Utilizing the high fluorescence response of labeled oligonucleotides, or the long phosphorescence lifetimes of transition metal complexes, we envision that our methodology could be adapted for the in vitro detection of disease-related factors in biological media such as human cellular and nuclear extracts. In order to maximize the selectivity, sensitivity and response time of our test, we will rigorously optimize the DNA sequence and the experimental parameters of the proposed assay.

We have already demonstrted that our approach gives satisfactory results. Extension to other systems is under way.

Our proposed methodologies are highly simple, sensitive, rapid, selective, low-cost and amenable to real-time and high-throughput analysis. We anticipate that the proposed technology could be highly useful to both academic and biomedical researchers worldwide working in the fields of medical diagnostics or biomarker biochemistry, and could be potentially applied for the “on-site” clinical diagnosis of human diseases.

5 articles in common between the two groups

Mutations in the genetic information of an organism can lead to the aberrant production of gene products. In humans, such genetic abnormalities can contribute to the development of various diseases. One type of DNA mutation is gene deletion, where a section of DNA or an entire part of a chromosome is missing. Some mutations can lead to genetic diseases characterized by elevated concentrations of biomarkers, which are usually proteins with critical roles in regulating cellular processes. The aberrant activity of such factors has been implicated in the development of a number of diseases, including cancer, developmental disorders and inflammation. Due to the importance of these agents as indicators and causative agents of diseases, their highly sensitive and efficient detection is of paramount importance for the early monitoring and therefore prevention and/or treatment of genetic diseases.

The development of small and portable probe for point-of-care diagnosis has emerged as a critical issue in in vitro diagnostics technology. To date, numerous methods have been developed for the selective detection of DNA, including the polymerase chain reaction and rolling circle amplification. However, these methods tend to be time-consuming, and involve tedious sample preparation and/or complex operation. For protein detection, the most commonly-used techniques include enzyme-linked immunosorbent assays (ELISA), capillary electrophoresis, and mass spectrometry. These detection methods typically involve multiple sample preparation steps and/or expensive instrumentation. Most importantly, the aforementioned detection methods generally require off-site laboratory equipment, and are not amenable to real-time point-of-care analysis.

We propose to develop "split-quadruplex" oligonucleotide-based sensing platforms for the detection of DNA mutations and protein biomarkers. Utilizing the high fluorescence response of labeled oligonucleotides, or the long phosphorescence lifetimes of transition metal complexes, we envision that our methodology could be adapted for the in vitro detection of disease-related factors in biological media such as human cellular and nuclear extracts. In order to maximize the selectivity, sensitivity and response time of our test, we will rigorously optimize the DNA sequence and the experimental parameters of the proposed assay. Our proposed methodologies are highly simple, sensitive, rapid, selective, low-cost and amenable to real-time and high-throughput analysis. We anticipate that the proposed technology could be highly useful to both academic and biomedical researchers worldwide working in the fields of medical diagnostics or biomarker biochemistry, and could be potentially applied for the “on-site” clinical diagnosis of human diseases.