Development of Ultra-fast Assays for RNA-based Disease Targets

Nucleic acid detection and amplification technologies have recently gained a lot of public attention for the application of pathogen detection. Therefore, research has been directed towards improving their speed, sensitivity and accuracy to achieve more reliable results and simpler, more applicable methodologies. Our research will focus on contributing to this field by optimising and further developing RTF-EXPAR (Carter et al., 2019.) method for RNA pathogen detection. RTF-EXPAR is an unique method which surpasses the lengthy reverse transcription (RT step) of RNA to DNA conversion by utilising specific complementary oligonucleotide and nicking restriction enzyme instead of the reverse transcriptase. Coupling this novel technology to a rapid nucleic acid amplification method, such as EXPAR, achieved SARS-CoV-2 RNA detection within 10 minutes. We aim to further develop this method to be applicable to a range of RNA pathogens, as well as nucleic-acid cancer biomarkers. Furthermore, we aim to explore nucleic acid detection read-out signal methods, other than the currently used intercalating fluorescence dyes, to develop a multiplexed assay for automated high-through put DNA/RNA target screening.