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期刊论文
Real-time detection of telomerase activity utilizing duplex scorpionprimers and hairpin-like reverse primers
New J.Chem., 2004, 28, 1488-1493,-0001,():
Telomerase, a ribonucleoprotein enzyme, is expected to be a new marker for cancer diagnosis. Thedevelopment of the telomeric repeat amplification protocol (TRAP) and its modified versions have facilitatedthe detection of telomerase activity in small tissue and tumour samples. But most of these techniques requirecomplex post-PCR procedures. As for the two real-time quantitative methods (SYBR Green and Amplifluormethods) reported so far, both use fluorogenic probes without specificity. To overcome these problems wedeveloped a new real-time method for the detection of telomerase activity. In this method a duplex scorpionprimer and reverse primers with hairpin-like structures were used. The use of duplex scorpion primers showsa series of advantages: the target-specific probe sequence provides higher specificity than the SYBR Greenand Amplifluor methods; the unimolecular probing mechanism allow the assay to be conducted under fastcycling conditions and a single operation can be completed in 1.5h; the closed-tube system reduces the riskof carryover contamination and supports high throughput. This method may be a useful tool to rapidly,specifically and precisely quantify telomerase activity.
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