Polymerase chain reaction-based enzyme immunoassay for quantitation of telomerase activity: application to colorectal cancers.
Telomerase is a specialized reverse transcriptase that synthesizes telomeric sequences onto human chromosomal ends. It appears to be present in the majority of primary human cancer tissues, and may have potential as a universal tumor marker. In this report, we describe a sensitive, non-radioactive, polymerase chain reaction (PCR)-based enzyme immunoassay (EIA) for the quantitation of telomerase activity in human cells. This PCR-EIA is convenient and can be easily completed within 3 h. The correlation coefficient between the results of PCR-EIA and the conventional telomeric repeat amplification protocol (TRAP) method, as measured on 4 different cell lines, was over 0.98. Evaluation of this method for clinical application was conducted with tissues obtained from patients with colorectal cancers and the results were compared with those of the conventional TRAP method. Our data indicate that telomerase activities measured by conventional TRAP and PCR-EIA are highly correlated, and we suggest that the PCR-EIA method can substitute for conventional TRAP.
Cheng
A J
AJ
Department of Medical Technology, Chang Gung University, Tao-Yuan, Taiwan.
Tang
R
R
Wang
J Y
JY
Chang
J T
JT
Wang
T C
TC
D001345:Autoradiography; D014408:Biomarkers, Tumor / Q000032:analysis; D002457:Cell Extracts; D015179:Colorectal Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007413:Intestinal Mucosa / Q000201:enzymology / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D012680:Sensitivity and Specificity; D013223:Statistics as Topic; D019098:Telomerase / Q000378:metabolism*; D013997:Time Factors; D017186:Titrimetry; D014407:Tumor Cells, Cultured
D014408:Biomarkers, Tumor; D002457:Cell Extracts; D019098:Telomerase; D012259:Ribonuclease, Pancreatic