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Clinical Chemistry 49: 1058-1065, 2003; 10.1373/49.7.1058
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(Clinical Chemistry. 2003;49:1058-1065.)
© 2003 American Association for Clinical Chemistry, Inc.

DNA Integrity as a Potential Marker for Stool-based Detection of Colorectal Cancer

Kevin A. Boynton1, Ian C. Summerhayes2, David A. Ahlquist3 and Anthony P. Shuber1,a

1 Applied Research Group, EXACT Sciences Corporation, Maynard, MA 01754.

2 Cell and Molecular Biology, R.E. Wise Research & Education Institute, Lahey Clinic, Burlington, MA 01805.

3 Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN 55905.

aAddress correspondence to this author at: EXACT Sciences Corporation, 63 Great Road, Maynard, MA 01754. Fax 978-897-3481; e-mail tshuber{at}exactsciences.com.

Background: Molecular genetic analysis of DNA in patient stools has been proposed for screening of colorectal cancer (CRC). Because nonapoptotic cells shed from tumors may contain DNA that is less degraded than DNA fragments from healthy colonic mucosa, our aim was to show that DNA fragments isolated from stools of patients with CRC had higher integrity than DNA isolated from stools of patients with healthy colonic mucosa.

Methods: We purified DNA from the stools of a colonoscopy-negative control group and patients with CRC and examined the relationship between long DNA fragments and clinical status by determining stool DNA integrity, using oligonucleotide-based hybrid captures with specific target sequences in increasingly long PCR reactions (200 bp, 400 bp, 800 bp, 1.3 kb, 1.8 kb, 24 kb). DNA fragments obtained from CRC patients were compared with fragments obtained from colonoscopy-negative individuals for length and/or integrity.

Results: DNA fragments isolated from CRC patients were of higher molecular weight (>18 bands detected of a total of 24 possible bands) than fragments isolated from fecal DNA of the colonoscopy-negative control group.

Conclusions: The presence of long DNA fragments in stool is associated with CRC and may be related to disease-associated differences in the regulation of proliferation and apoptosis. An assay of fecal DNA integrity may be a useful biomarker for the detection of CRC.




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