Citation: Xiao Ming Zhou, Li Jia. Detection of telomerase activity by combination of telomeric repeat amplification protocol and electrochemiluminescence assay[J]. Chinese Chemical Letters, ;2008, 19(6): 699-702. doi: 10.1016/j.cclet.2008.04.017 shu

Detection of telomerase activity by combination of telomeric repeat amplification protocol and electrochemiluminescence assay

Xiao Ming Zhou ,
Li Jia

MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University, Guangzhou 510631, China

Received Date: 25 December 2007

Fund Project: This research is supported by the National Natural Science Foundation of China (No. 30600128the National High Technology Research and Development Program of China (863 Program) (No. 2007AA10Z204)and the Natural Science Foundation of Guangdong Province (No. 7005825). 30700155)

A highly sensitive telomerase detection method that combines telomeric repeat amplification protocol (TRAP) and magnetic beads based electrochemiluminescence (ECL) assay has been developed. Briefly, telomerase recognizes biotinylated telomerase synthesis primer (B-TS) and synthesizes extension products, which then serve as the templates for PCR amplification using B-TS as the forward primer and Iris-(2´2´-bipyridyl) ruthenium (TBR) labeled ACX (TBR-ACX) as the reversed primer. The amplified product is captured on streptavidin-coated paramagnetic beads and detected by ECL. Telomerase positive HeLa cells were used to validate the feasibility of the method. The experimental results showed down to 10 cancer cells can be detected easily. The method is a useful tool for telomerase activity analysis due to its sensitivity, rapidity, safety, high throughput, and low cost. It can be used for screening a large amount of clinical samples.
- Telomerase activity,
- TRAP,
- Electrochemiluminescence,
- Cancer diagnosis
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