Visual and sensitive detection of telomerase activity via hydrogen peroxide test strip

The point of care testing (POCT) of telomerase activity is critical for early diagnosis of cancer. Herein, a colorimetric method was developed for visual detection of telomerase activity via hydrogen peroxide test strip. It is based on the telomerase-controlled in-situ formation of hydrogen peroxide. Firstly, biotinylated telomerase substrate (TS) primer was attached on the surface of magnetic beads (MBs) via the streptavidin-biotin reaction to form MB-TS complex. Then, TS primers were elongated by telomerase to form long telomere elongated products (TEP) which contains TTAGGG repeat units. The in-situ formed MB-TEP complex specifically hybridized with glucose oxidase modified cDNA (GOD-cDNA). After magnetic separation and washing, the MB-TEP/GOD-cDNA complex incubated with glucose solution to in-situ produce hydrogen peroxide which was detected by hydrogen peroxide test strip. One long TEP hybridized with multiple GOD-cDNAs, which enriched GOD to highly efficiently catalyze glucose for generating hydrogen peroxide. Thus, the visual assay achieved sensitive detection of telomerase activity, and the limit of detection (LOD) reached as low as 10 HeLa cells/μL by naked eyes and 4.5 HeLa cells/μL by absorbance measurements. Therefore, it offers a sensitive and low-cost method for visual detection of telomerase activity, which also, widens the application of commercial hydrogen peroxide test strip in the development of non-H₂O₂ biosensors.