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International Journal of Colorectal Disease

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01-05-2011 | Original Article

A simplified, non-invasive fecal-based DNA integrity assay and iFOBT for colorectal cancer detection

Authors: Murugan Kalimutho, Giovanna Del Vecchio Blanco, Micaela Cretella, Elena Mannisi, Pierpaolo Sileri, Amanda Formosa, Francesco Pallone, Giorgio Federici, Sergio Bernardini

Published in: International Journal of Colorectal Disease | Issue 5/2011

Abstract

Purpose

Neoplasia cells exfoliated from colorectal epithelium have dysfunctional apoptotic mechanisms, thus it is possible to identify high-molecular weight DNA fragments in feces. This prospective single-center study was performed to evaluate the sensitivity and specificity of fecal-based DNA integrity versus immunological fecal occult blood test (iFOBT) and calprotectin for colorectal cancer (CRC) and adenoma detection.

Methods

Feces were collected from 204 subjects and DNA integrity was quantified by quantitative-denaturing high performance liquid chromatography (QdHPLC). Calprotectin and iFOBT were assessed using commercial kits. The diagnostic performance was calculated by receiver operating characteristic (ROC) curves analysis.

Results

A total of 192 fecal specimens were analyzed and 12 samples were excluded due to DNA degradation. We found long DNA (L-DNA) occurrence in feces with a sensitivity of 86% (n = 24/28) and a specificity of 81% for CRC detection. To minimize false-positive cases of the developed test, area under the curve of ROC was evaluated such that the specificity was increased to 92% with decreased sensitivity to 79%, p = 0.0001 for CRC detection. iFOBT was positive in 51% (n = 14/27) while calprotectin was positive in 75% (n = 18/27). The combination of iFOBT and L-DNA identified a greater number of CRC cases with a sensitivity of 89% and a specificity of 95%, p < 0.001. The combination also improved the sensitivity of polyps, particularly high-grade dysplasia and advanced adenoma (33%, p = 0.0015) as opposed to a single evaluation assay (17–21%).

Conclusions

This study illustrates the usefulness of fecal DNA integrity assay by QdHPLC as a non-invasive, easy-to-perform, and reproducible method with a high level of sensitivity in detecting individuals with colorectal neoplasia. Combination of iFOBT and L-DNA improves the sensitivity for CRC and adenoma detection.

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Ratio is referred to the normalization of the detected L-DNA by QdHPLC over β-actin PCR amplification on the same patient

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Title: A simplified, non-invasive fecal-based DNA integrity assay and iFOBT for colorectal cancer detection
Authors: Murugan Kalimutho
Giovanna Del Vecchio Blanco
Micaela Cretella
Elena Mannisi
Pierpaolo Sileri
Amanda Formosa
Francesco Pallone
Giorgio Federici
Sergio Bernardini
Publication date: 01-05-2011
Publisher: Springer-Verlag
Published in: International Journal of Colorectal Disease / Issue 5/2011
Print ISSN: 0179-1958
Electronic ISSN: 1432-1262
DOI: https://doi.org/10.1007/s00384-010-1128-x

Keynote webinar | Spotlight on medication adherence

Springer Medicine

A simplified, non-invasive fecal-based DNA integrity assay and iFOBT for colorectal cancer detection

Abstract

Purpose

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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