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Regular version of the site

Article

Novel homo Yin-Yang probes improve sensitivity in RT-qPCR detection of low copy HIV RNA

Talanta. 2019. Vol. 194. P. 226-232.
Vladimir A. Korshun.

Nucleic acids labeled with a fluorophore/quencher pair are widely used as probes in biomedical research and
molecular diagnostics. Here we synthesized novel DNA molecular beacons double labeled with the identical dyes
(R6G, ROX and Cy5) at 5′- and 3′-end and studied their photo physical properties. We demonstrated that
fluorescence quenching by formation of the homo dimer exciton in such molecular beacons allows using them in
homogeneous assays. Further, we developed and evaluated homo Yin-Yang DNA probes labeled with identical
dyes and used them for detection of low copy HIV RNA by RT-qPCR. They demonstrated improved sensitivity
(LLQ: 10 vs 30 copies mL-1) in comparison to commercially available Abbott RealTime HIV-1 kit based on VICBHQ
dyes both for model mixtures (naive human plasma with added deactivated HIV-1 virus) and for preliminarily
confirmed 36 clinical samples (4 vs 1 positive ones for low-copy samples).