Absolute Quantitative Real-Time Polymerase Chain Reaction For The Measurement of Human Papillomavirus E7 mRNA In Cervical Cytobrush Specimens

Authors

Michael E. Scheurer, The University of Texas M. D. Anderson Cancer Center, Department of Epidemiology, Houston, Tx
Laura M. Dillon, The University of Texas Dental Branch, Department of Diagnostic Sciences, Houston, Tx
Zhuo Chen, Emory University Winship Cancer Institute, Department of Hematology and Oncology
Michele Follen, The University of Texas M. D. Anderson Cancer Center, Department of Gynecologic Oncology and Biomedical Engineering Center, Houston, Tx
Karen Adler-Storthz, The University of Texas Dental Branch, Department of Diagnostic Sciences, Houston, TxFollow

Publication Date

1-1-2007

Journal

Infectious Agents and Cancer

Abstract

BACKGROUND: Few reports of the utilization of an accurate, cost-effective means for measuring HPV oncogene transcripts have been published. Several papers have reported the use of relative quantitation or more expensive Taqman methods. Here, we report a method of absolute quantitative real-time PCR utilizing SYBR-green fluorescence for the measurement of HPV E7 expression in cervical cytobrush specimens. RESULTS: The construction of a standard curve based on the serial dilution of an E7-containing plasmid was the key for being able to accurately compare measurements between cervical samples. The assay was highly reproducible with an overall coefficient of variation of 10.4%. CONCLUSION: The use of highly reproducible and accurate SYBR-based real-time polymerase chain reaction (PCR) assays instead of performing Taqman-type assays allows low-cost, high-throughput analysis of viral mRNA expression. The development of such assays will help in refining the current screening programs for HPV-related carcinomas.

Comments

PMCID: PMC1852093

DOI

10.1186/1750-9378-2-8

PMID

17407544

PMCID

PMC1852093

PubMedCentral® Posted Date

April 2007

PubMedCentral® Full Text Version

Post-print

Recommended Citation

Scheurer, Michael E.; Dillon, Laura M.; Chen, Zhuo; Follen, Michele; and Adler-Storthz, Karen, "Absolute Quantitative Real-Time Polymerase Chain Reaction For The Measurement of Human Papillomavirus E7 mRNA In Cervical Cytobrush Specimens" (2007). Faculty, Staff and Student Publications. 8.
https://digitalcommons.library.tmc.edu/uthdb_docs/8