New method to measure telomerase activity by transcription-mediated amplification and hybridization protection assay.
Telomerase is a ribonucleoprotein complex that uses RNA as a template for the addition of telomeric repeats. The development of the telomeric repeat amplification protocol (TRAP), a sensitive PCR-based assay, has facilitated the detection of telomerase activity in small tissue and tumor samples. Telomerase activity is expected to be a new diagnostic and prognostic marker of human cancer. In this study, we applied a non-PCR-based transcription-mediated amplification (TMA) and hybridization protection assay (HPA) to the measurement of telomerase activity by modification of both primers in TMA. We demonstrated that the modified TMA can detect and measure telomerase activity. TMA/HPA is as sensitive and reproducible as conventional TRAP, but is both faster and easier to perform. Furthermore, we found that TMA/HPA was influenced minimally by TRAP inhibitors that may come from clinical samples. TMA/HPA, which is easy, rapid, and applicable to a high-throughput format, should be clinically useful for the detection and monitoring of telomerase activity.
Hirose
M
M
1 Diagnostics Science Laboratory, Chugai Diagnostics Science Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171-8545, Japan. hirosemnr@chungai-pharm.co.jp
Abe-Hashimoto
J
J
Tahara
H
H
Ide
T
T
Yoshimura
T
T
D006528:Carcinoma, Hepatocellular / Q000201:enzymology; D015179:Colorectal Neoplasms / Q000201:enzymology; D017931:DNA Primers; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D019098:Telomerase