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Rapid Quantification of Telomerase Activity Employing an Improved Real-time Telomeric Repeat Amplification Protocol in Clinical Tissue Samples Eliminates Interference by PCR Inhibitors | Abstract
ISSN: 1948-5956

Journal of Cancer Science & Therapy
Open Access

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Research Article

Rapid Quantification of Telomerase Activity Employing an Improved Real-time Telomeric Repeat Amplification Protocol in Clinical Tissue Samples Eliminates Interference by PCR Inhibitors

Philipp Schriefer1, Cagatay Günes2 and Henning Wege3*

1Department of Urology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg

2Institute of Molecular Medicine and Max-Planck-Research Group for Stem Cell Aging, University of Ulm, Albert-Einstein-Allee 11, D-89081 Ulm

3Department of Gastroenterology and Hepatology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg

*Corresponding Author:
Dr. Henning Wege
Department of Gastroenterology and Hepatology
University Medical Center Hamburg-Eppendorf
Martinistrasse 52
D-20246 Hamburg, Germany
Tel: + 49 152 22816347
Fax: +49 40 7410 59092
E-mail:
[email protected]

Received Date: September 30, 2011; Accepted Date: October 16, 2011; Published Date: October 18, 2011

Citation: Schriefer P, Günes C, Wege H (2011) Rapid Quantification of Telomerase Activity Employing an Improved Real-time Telomeric Repeat Amplification Protocol in Clinical Tissue Samples Eliminates Interference by PCR Inhibitors. J Cancer Sci Ther 3: 176-180. doi:10.4172/1948-5956.1000084

Copyright: © 2011 Schriefer P, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Telomerase, a ribonucleoprotein with reverse transcriptase activity, enables human cells to maintain chromosomal stability and to proliferate without limits. Various studies demonstrated telomerase activation in human cancer, including hepatocellular carcinoma. Therefore, quantification of telomerase activity has been proposed as diagnostic and prognostic tool. In this study, we optimized a 1-step real-time quantitative telomeric repeat amplification protocol for the robust and rapid quantification of telomerase activity in clinical tissue samples. To ensure undisturbed PCR kinetics even in samples with high telomerase activity, we initially determined optimal sample dilution for our assay. Next, we assessed highly diluted samples and did not observe relevant interference by tissue inhibitors of PCR, which constitute a major problem analyzing clinical tissue samples with end-point assays. To test our real-time assay, we evaluated human liver samples and detected increased telomerase activity in malignant liver lesions, whereas benign liver tissue displayed only minimal telomerase activity. In conclusion, our optimized assay is suitable to quantify telomerase activity in clinical tissue samples without interference by PCR inhibitors. The assay may be employed to detect telomerase activity during carcinogenesis and to monitor telomerase activity during cancer progression and treatment.

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