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

Authors

Language

English

Publication Date

1-1-2007

Journal

Infectious Agents and Cancer

DOI

10.1186/1750-9378-2-8

PMID

17407544

PMCID

PMC1852093

PubMedCentral® Posted Date

April 2007

PubMedCentral® Full Text Version

Post-print

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.

Recommended Citation

, "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