Sensitive detection of colorectal cancer in peripheral blood by septin 9 DNA methylation assay

PLoS One. 2008;3(11):e3759. doi: 10.1371/journal.pone.0003759. Epub 2008 Nov 19.

Abstract

Background: Colorectal cancer (CRC) is the second leading cause of cancer deaths despite the fact that detection of this cancer in early stages results in over 90% survival rate. Currently less than 45% of at-risk individuals in the US are screened regularly, exposing a need for better screening tests. We performed two case-control studies to validate a blood-based test that identifies methylated DNA in plasma from all stages of CRC.

Methodology/principal findings: Using a PCR assay for analysis of Septin 9 (SEPT9) hypermethylation in DNA extracted from plasma, clinical performance was optimized on 354 samples (252 CRC, 102 controls) and validated in a blinded, independent study of 309 samples (126 CRC, 183 controls). 168 polyps and 411 additional disease controls were also evaluated. Based on the training study SEPT9-based classification detected 120/252 CRCs (48%) and 7/102 controls (7%). In the test study 73/126 CRCs (58%) and 18/183 control samples (10%) were positive for SEPT9 validating the training set results. Inclusion of an additional measurement replicate increased the sensitivity of the assay in the testing set to 72% (90/125 CRCs detected) while maintaining 90% specificity (19/183 for controls). Positive rates for plasmas from the other cancers (11/96) and non-cancerous conditions (41/315) were low. The rate of polyp detection (>1 cm) was approximately 20%.

Conclusions/significance: Analysis of SEPT9 DNA methylation in plasma represents a straightforward, minimally invasive method to detect all stages of CRC with potential to satisfy unmet needs for increased compliance in the screening population. Further clinical testing is warranted.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Algorithms
  • Case-Control Studies
  • Colorectal Neoplasms / blood*
  • Colorectal Neoplasms / diagnosis*
  • Colorectal Neoplasms / genetics
  • DNA Methylation*
  • Female
  • GTP Phosphohydrolases / blood
  • GTP Phosphohydrolases / genetics*
  • Humans
  • Male
  • Mass Screening / methods*
  • Middle Aged
  • Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Septins
  • Treatment Outcome

Substances

  • GTP Phosphohydrolases
  • SEPTIN9 protein, human
  • Septins