Tandem Oligonucleotide Probe Annealing and Elongation To Discriminate Viral Sequence

摘要

New approaches for genomic DNA/RNA detection are in high demand in order to provide controls for existing enzymatic technologies and to create alternatives for emerging applications. In particular, there is an unmet need in rapid, reliable detection of short RNA regions which could open up new opportunities- in transcriptome analysis, virology, and other fields. Herein, we report for the first:time a "click" chemistry approach to oligonucleotide probe elongation as a novel approach to specifically detect a viral sequence. We hybridized a library of short, terminally-labeled probes to Ebola virus RNA followed by click assembly and analysis of the read seqUence by various 'techniques. As we demonstrate in this paper, using our new approach, a viral RNA sequence can be detected in less than 2 h without the need for cDNA synthesis or any other enzymatic reactions and with a sensitivity of <10 pM target RNA.