Quantification of total plasma cell-free DNA in ovarian cancer using real-time PCR

Aparna A. Kamat, Anil K. Sood, Dianne Dang, David M. Gershenson, Joe L. Simpson, Farideh Z. Bischoff

Research output: Chapter in Book/Report/Conference proceedingConference contribution

46 Scopus citations

Abstract

Our objective was to compare the levels of total circulating plasma cell-free DNA (CfDNA) using real-time PCR in patients with late-stage ovarian cancer with those in unaffected controls. Following IRB consent, DNA was extracted from archived frozen plasma of 19 patients with primary ovarian carcinoma and 12 age-matched controls using Qiagen DNA Isolation Kits. Quantification of total CfDNA was performed using real-time PCR with the TaqMan Assay for GAPDH, β-actin and β-globin and the number of genome equivalents (GE/mL) were determined from a standard curve. CfDNA levels of these loci were compared between the groups with Student's t-test, with P < 0.05 being statistically significant. The mean age of the patients was 61.6 years (±9.6) and of the controls was 54 years (±12.2). All patients had high-grade, advanced stage (III or IV) serous ovarian carcinomas. Preoperative CA-125 levels ranged from 43 to 15,626 IU/mL (mean 2487.2 ± 3686 IU/mL). Total CfDNA in ovarian cancer was higher among patients with ovarian cancer as compared to controls at all three loci: GAPDH (P = 0.022), β-actin (P = 0.025), and β-globin (P = 0.0089). CfDNA is elevated in advanced stage disease compared to controls. These preliminary results suggest that total CfDNA in the plasma of patients with ovarian cancer may be useful For noninvasive screening and disease surveillance.

Original languageEnglish (US)
Title of host publicationCirculating Nucleic Acids in Plasma and Serum IV
PublisherBlackwell Publishing Inc.
Pages230-234
Number of pages5
ISBN (Print)157331627X, 9781573316279
DOIs
StatePublished - Sep 2006

Publication series

NameAnnals of the New York Academy of Sciences
Volume1075
ISSN (Print)0077-8923
ISSN (Electronic)1749-6632

Keywords

  • Cell-free DNA
  • Ovarian cancer
  • Quantification
  • Real-time PCR

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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