Fast and non-PCR Detection of RNAs based on Molecular Fishing

Description

This project aims to develop a highly versatile and powerful technique for rapid, non-PCR, highly sensitive detection of RNA from virus, including SARS-Cov-2. The proposed testing strategy will be capable of cherry-picking targeted signature RNAs from liquid sample prepared from SARS-CoV-2 viruses. The technique combines “molecular fishing” that have been continuously developed by the research team using diamond nanoneedles for manipulating cellular systems and for probing various biological markers. Based on these works, the proposed platform for the quantitative detection of SARS-Cov-2 will have a detection limit as low as 100 attomolar, corresponding to only 50 copies of RNAs per microliter, and a turnover time of ~15 minutes per assay. Notably, a novel RNA processing protocol will be employed to address the unpredictable degradation issue in existing methods, so that the SARS-CoV-2 signature RNAs could be converted to fragments of 18-25 nucleotides, and then be captured and enriched by diamond nanoneedles functionalized with p19 RNA biding proteins, thus reducing the false-negative and false-positive possibility.