title: Absolute quantitative real-time polymerase chain reaction for the measurement of human papillomavirus E7 mRNA in cervical cytobrush specimens.
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%.
  
  
  CONCLUSIONS
  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.
authors.affiliation: The University of Texas M. D. Anderson Cancer Center, Department of Epidemiology, Unit 1340, PO Box 301439, Houston, TX 77230-1439, USA. mescheur@mdanderson.org
authors.forename: Michael E
authors.initials: ME
authors.lastname: Scheurer
authors.forename: Laura M
authors.initials: LM
authors.lastname: Dillon
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Chen
authors.forename: Michele
authors.initials: M
authors.lastname: Follen
authors.forename: Karen
authors.initials: K
authors.lastname: Adler-Storthz