Real-time PCR detection of telomerase activity using specific molecular beacon probes.
Telomerase is a potentially important biomarker and a prognostic indicator of cancer. Several techniques for assessing telomerase activity, including the telomeric repeat amplification protocol (TRAP) and its modified versions, have been developed. Of these methods, real-time quantitative TRAP (RTQ-TRAP) is considered the most promising. In this work, a novel RTQ-TRAP method is developed in which a telomeric repeats-specific molecular beacon is used. The use of the molecular beacon can improve the specificity of the RTQ-TRAP assay, making the method suitable for studying the overall processivity results and the turnover rate of telomerase. In addition, the real-time, closed-tube protocol used obviates the need for post-amplification procedures, reduces the risk of carryover contamination, and supports high throughput. Its performance in synthetic telomerase products and cell extracts suggests that the developed molecular beacon assay can further enhance the clinical utility of telomerase activity as a biomarker/indicator in cancer diagnosis and prognosis. The method also provides a novel approach to the specific detection of some particular gene sequences to which sequence-specific fluorogenic probes cannot be applied directly. Figure Real-time PCR detection of telomerase activity using specific molecular beacon probes.
Kong
Deming
D
Key Laboratory of Functional Polymer Materials, Ministry of Education, Nankai University, Tianjin, 300071, China. kongdem@nankai.edu.cn
Jin
Yawei
Y
Yin
Yuji
Y
Mi
Huaifeng
H
Shen
Hanxi
H
D014408:Biomarkers, Tumor / Q000032:analysis*; D018922:HL-60 Cells; D006801:Humans; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D019098:Telomerase / Q000032:analysis*
D014408:Biomarkers, Tumor; D015335:Molecular Probes; D019098:Telomerase