Validation of a real-time PCR-based qualitative assay for the detection of methylated SEPT9 DNA in human plasma

Nicholas Potter, Patrick Hurban, Mary N. White, Kara D. Whitlock, Catherine E. Lofton-Day, Reimo Tetzner, Thomas Koenig, Neil B. Quigley, Gunter Weiss

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

BACKGROUND: Epi proColon® is a new blood-based colorectal cancer (CRC) screening test designed to determine the methylation status of a promoter region of the SEPT9 (septin 9) gene in cell-free DNA isolated from plasma. We describe the analytical and clinical performance of the test. METHODS: Analytical performance at 4 testing laboratories included determination of limit of detection, precision, and reproducibility of the SEPT9 test. Clinical performance was evaluated in a prospective study by use of samples (n = 1544) from subjects enrolled in the PRESEPT clinical trial. Results were analyzed by comparison with colonoscopy, the reference standard. RESULTS: The limit of detection for methylated SEPT9 DNA was 7.8 pg/mL (95% CI 6-11 pg/mL) corresponding to <2 genome copies of methylated SEPT9 per milliliter of plasma. In the prospective clinical trial, sensitivity for all stages of CRC was 68% (95% CI 53%-80%) and for stage I-III CRC, 64% (48%-77%). Adjusted specificity, on the basis of negative colonoscopy findings, was 80.0% (78%-82%). SIGNIFICANCE: The Epi proColon test is a simple, realtime PCR-based assay for the detection of methylated SEPT9 DNA in blood that may provide a noninvasive CRC screening alternative for people noncompliant with current CRC screening guidelines.

Original languageEnglish (US)
Pages (from-to)1183-1191
Number of pages9
JournalClinical Chemistry
Volume60
Issue number9
DOIs
StatePublished - Jan 1 2014

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Septins
Plasma (human)
Real-Time Polymerase Chain Reaction
Colorectal Neoplasms
Assays
Early Detection of Cancer
DNA
Screening
Colonoscopy
Limit of Detection
Blood
Genes
Clinical Trials
Plasmas
Methylation
Genetic Promoter Regions
Genome
Prospective Studies
Guidelines
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Validation of a real-time PCR-based qualitative assay for the detection of methylated SEPT9 DNA in human plasma. / Potter, Nicholas; Hurban, Patrick; White, Mary N.; Whitlock, Kara D.; Lofton-Day, Catherine E.; Tetzner, Reimo; Koenig, Thomas; Quigley, Neil B.; Weiss, Gunter.

In: Clinical Chemistry, Vol. 60, No. 9, 01.01.2014, p. 1183-1191.

Research output: Contribution to journalArticle

Potter, N, Hurban, P, White, MN, Whitlock, KD, Lofton-Day, CE, Tetzner, R, Koenig, T, Quigley, NB & Weiss, G 2014, 'Validation of a real-time PCR-based qualitative assay for the detection of methylated SEPT9 DNA in human plasma', Clinical Chemistry, vol. 60, no. 9, pp. 1183-1191. https://doi.org/10.1373/clinchem.2013.221044
Potter, Nicholas ; Hurban, Patrick ; White, Mary N. ; Whitlock, Kara D. ; Lofton-Day, Catherine E. ; Tetzner, Reimo ; Koenig, Thomas ; Quigley, Neil B. ; Weiss, Gunter. / Validation of a real-time PCR-based qualitative assay for the detection of methylated SEPT9 DNA in human plasma. In: Clinical Chemistry. 2014 ; Vol. 60, No. 9. pp. 1183-1191.
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