Simultaneous Detection of Colorectal Cancer Mutations in Stool Samples with Biochip Arrays

Helena Murray,Damien Mcaleer,Andrew Cartwright,Mark J Latten ,S Fitzgerald

doi:10.2478/v10011-009-0028-5

Abstract

Simultaneous Detection of Colorectal Cancer Mutations in Stool Samples with Biochip ArraysColorectal cancer (CRC) is the second main cause of cancer-related death in the Western world and like many other tumours is curable if detected at an early stage. Current detection options include faecal occult blood testing and invasive direct visualisation techniques such as flexible sigmoidoscopy, colonoscopy and barium enema. The availability of a more simple, non-invasive test that detects tumour specific products with optimal analytical performance might overcome barriers among patients who are not willing to undergo more sensitive but invasive tests. One such emerging technology, which has shown promise in recent years, is the analysis of DNA alterations exfoliated from tumour cells into stool. Here we report an analytical platform for non-invasive detection of 28 common mutations within CRC-related genesAPC, TP53, K-rasandBRAFin stool samples based on biochip array technology and applied to the semi-automated Evidence Investigator analyser. Mutation detection was possible in 1000-fold excess of wildtype DNA and analysis of 10 CRC-positive patient samples showed presence of targeted mutations with equivalent mutations also identified by an alternative method. This application represents an excellent tool for the multiplex detection of CRC-specific mutations using a single platform.

Highlights

Colorectal cancer (CRC) has a long premalignant phase and relatively slow progression from organ defined invasive disease to local and distant metastatic disease and as such there is ampleAddress for correspondence: Mr Damien McAleer Randox Laboratories Ltd. 55 Diamond Road, Crumlin Co
Summary: Colorectal cancer (CRC) is the second main cause of cancer-related death in the Western world and like many other tumours is curable if detected at an early stage
The availability of a more simple, non-invasive test that detects tumour specific products with optimal analytical performance might overcome barriers among patients who are not willing to undergo more sensitive but invasive tests. One such emerging technology, which has shown promise in recent years, is the analysis of DNA alterations exfoliated from tumour cells into stool

Summary

Introduction

Colorectal cancer (CRC) has a long premalignant phase and relatively slow progression from organ defined invasive disease to local and distant metastatic disease and as such there is ampleAddress for correspondence: Mr Damien McAleer Randox Laboratories Ltd. 55 Diamond Road, Crumlin Co. Kratak sadraj: Kolorektalni kancer je drugi glavni uzrok smrtnosti usled raka u zapadnom svetu i poput mnogih drugih tumora izle~iv je ukoliko se otkrije u ranoj fazi. Dostupnost jednostavnijeg, neinvazivnog testa za otkrivanje produkata specifi~nih za tumor uz optimalne analiti~ke performanse doprinela bi prevazilaenju barijera kod pacijenata koji se nerado podvrgavaju senzitivnijim ali invazivnim testovima. Takva tehnologija koja se poslednjih godina razvija i daje ohrabruju}e rezultate je analiza alteracija DNK koje iz }elija tumora dospevaju u stolicu. Detekciju mutacija bilo je mogu}e uraditi u 1000-strukom vi{ku DNK a analiza 10 uzoraka pacijenata pozitivnih na kolorektalni kancer je otkrila prisustvo ciljanih mutacija sa ekvivalentnim mutacijama tako|e identifikovanim alternativnom metodom. Takva primena predstavlja sjajnu alatku za detekciju mutacija specifi~nih za kolorektalni kancer pomo}u jedinstvene platforme

Methods

Results

Discussion

Conclusion