Limitations on the quantitative determination of telomerase activity by the electrophoretic and ELISA based TRAP assays.
Telomerase is a promising new tumor marker and can be detected using the TRAP (Telomeric Repeat Amplification Protocol) method. To address factors affecting its quantitative determination, we evaluated two commercial TRAP assays, an electrophoretic and an ELISA assay formats, using cultured cells and human tumor samples. We found that both TRAP assays had a limited linearity from 250 to 5000 tumor cells, with a similar intra-assay variation. The quantification of TRAP products was affected by high cell number in sample, the presence of non-tumor cells, and interfering substances in patient specimens. Because both assays have different limitations, determination of telomerase by a combined use of the two may provide more accurate information on the telomerase activity in a specimen. Extracts of specimens should also be tested at several concentrations to insure that the result is not being falsely decreased by an inhibitor. The quantitative results for telomerase activity by the TRAP assays, however, should be interpreted cautiously.
Wu
Y Y
YY
Clinical Pathology Department, CC NIH, Building 10, Room 2C 407, Bethesda, MD 20892-1508, USA.
Hruszkewycz
A M
AM
Delgado
R M
RM
Yang
A
A
Vortmeyer
A O
AO
Moon
Y W
YW
Weil
R J
RJ
Zhuang
Z
Z
Remaley
A T
AT
D002452:Cell Count; D002793:Cholic Acids; D004586:Electrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D019098:Telomerase / Q000032:analysis*; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000201:enzymology
D002793:Cholic Acids; D019098:Telomerase; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate