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Infectious Agents and Cancer

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Open Access 01-12-2007 | Methodology

Absolute quantitative real-time polymerase chain reaction for the measurement of human papillomavirus E7 mRNA in cervical cytobrush specimens

Authors: Michael E Scheurer, Laura M Dillon, Zhuo Chen, Michele Follen, Karen Adler-Storthz

Published in: Infectious Agents and Cancer | Issue 1/2007

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.

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Title: Absolute quantitative real-time polymerase chain reaction for the measurement of human papillomavirus E7 mRNA in cervical cytobrush specimens
Authors: Michael E Scheurer
Laura M Dillon
Zhuo Chen
Michele Follen
Karen Adler-Storthz
Publication date: 01-12-2007
Publisher: BioMed Central
Published in: Infectious Agents and Cancer / Issue 1/2007
Electronic ISSN: 1750-9378
DOI: https://doi.org/10.1186/1750-9378-2-8

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