Sensitive High-Resolution Melting Analysis for Screening of KRAS and BRAF Mutations in Iranian Human Metastatic Colorectal Cancers

Document Type : Research Articles

Authors

1 Department of Virology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

2 Gastrointestinal and liver diseases research center, firoozgar hospital, Iran University of Medical Sciences. Tehran, Iran

3 Gastrointestinal and liver diseases research center, firoozgar hospital, Iran University of Medical Sciences, Tehran, Iran.

4 Department of pediatrics, shahid beheshti University of Medical Sciences, Tehran, Iran.

5 Department of Oncology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

 
Background: Investigations of methods for detection of mutations have uncovered major weaknesses of direct sequencing and pyrosequencing, with their high costs and low sensitivity in screening for both known and unknown mutations. High resolution melting (HRM) analysis is an alternative tool for the rapid detection of mutations. Here we describe the accuracy of HRM in screening for KRAS and BRAF mutations in metastatic colorectal cancer (mCRCs) samples. Materials and Methods: A total of 1000 mCRC patients in Mehr Hospital, Tehran, Iran, from Feb 2008 to May 2012 were examined for KRAS mutations and 242 of them were selected for further assessment of BRAF mutations by HRM analysis. In order to calculate the sensitivity and specificity, HRM results were checked by pyrosequencing as the golden standard and Dxs Therascreen as a further method. Results: In the total of 1,000 participants, there were 664 (66.4%) with wild type and 336 (33.6%) with mutant codons 12 and/or 13 of the KRAS gene. Among 242 samples randomly checked for the BRAF gene, all were wild type by HRM. Pyrosequencing and Dxs Therascreen results were in line with those of the HRM. In this regard, the sensitivity and specificity of HRM were evaluated as 100%. Conclusion: The findings suggest that the HRM, in comparison with DNA sequencing, is a more appropriate method for precise scanning of KRAS and BRAF mutations. It is also possible to state that HRM may be an attractive technique for the detection of known or unknown somatic mutations in other genes.

Keywords

Asian Pacific Journal of Cancer Prevention
Volume 17, Issue 12
December 2016
Pages 5147-5152

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History

  • Receive Date: 01 November 2016
  • Revise Date: 10 December 2016
  • Accept Date: 16 January 2017

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