引用本文:
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
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
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
Detection of telomerase activity by combination of telomeric repeat amplification protocol and electrochemiluminescence assay
摘要:
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.
English
Detection of telomerase activity by combination of telomeric repeat amplification protocol and electrochemiluminescence assay
Abstract:
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.
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Key words:
- Telomerase activity
- / TRAP
- / Electrochemiluminescence
- / Cancer diagnosis
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