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[Quantitation of telomerase activity using the polymerase chain reaction-based enzyme immunoassay and its clinical
Wei-qun Chen1, He-ming Chen, Yuan-jia Hu
1Department of Clinical Biochemistry, Xiangya School of Medicine, Central South University, Changsha 410013, China.
Objective:
To investigate detective methodology of telomerase activity and its clinical application.
Methods:
The telomerase activity was quantitated by polymerase chain reaction-base enzyme immunoassay (PCR-EIA). The TS primer was biotinylated (TS-B), and CX primer was digoxigeninated (CX-D). The amplicons containing TS-B were combined with microtiter plate coated streptavidin, then combined with anti-digoxigenin antibody labeled with POD and finally reacted to tetramethylbenzidine substrate solution.
Results:
Telomerase activity measured by the PCR-EIA method was comparable to that obtained from TRAP-silver stain protocol. The CV of the PCR-EIA method was 4.138%. Telomerase activity was detected in 90% of various tumer tissues. In control tissues, telomerase activity was detected only in 7.1%.
Conclusion:
The PCR-EIA method offers a rapid, quantitative, and nonisotopic assay for the determination of telomerase activity. It is simpler than silver stain protocol. The detection of telomerase activity may play a significant role in the diagnosis of clinical tumors.