[Quantitation of telomerase activity using the polymerase chain reaction-based enzyme immunoassay and its clinical application].
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%.


CONCLUSIONS
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.
Chen
Wei-qun
WQ
Department of Clinical Biochemistry, Xiangya School of Medicine, Central South University, Changsha 410013, China.
Chen
He-ming
HM
Hu
Yuan-jia
YJ
D015179:Colorectal Neoplasms / Q000201:enzymology; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods; D008175:Lung Neoplasms / Q000201:enzymology; D009303:Nasopharyngeal Neoplasms / Q000473:pathology; D009369:Neoplasms / Q000201:enzymology*; D016133:Polymerase Chain Reaction / Q000379:methods; D019098:Telomerase / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D019098:Telomerase