title: Sensitive High-Resolution Melting Analysis for Screening of KRAS and BRAF Mutations in Iranian Human Metastatic Colorectal Cancers
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.
authors.affiliation: Department of Virology, Faculty of Medicine,Shahid Beheshti University of Medical Sciences, Tehran, Iran. Email: n_rakhshani@yahoo.com
authors.forename: Mohammad Hadi
authors.initials: MH
authors.lastname: Karbalaie Niya
authors.forename: Ali
authors.initials: A
authors.lastname: Basi
authors.forename: Aghigh
authors.initials: A
authors.lastname: Koochak
authors.forename: Fahimeh
authors.initials: F
authors.lastname: Safarnezhad Tameshkel
authors.forename: Nasser
authors.initials: N
authors.lastname: Rakhshani
authors.forename: Farhad
authors.initials: F
authors.lastname: Zamani
authors.forename: Farid
authors.initials: F
authors.lastname: Imanzade
authors.forename: Hamid
authors.initials: H
authors.lastname: Rezvani
authors.forename: Mohammad Mahdi
authors.initials: MM
authors.lastname: Adib Sereshki
authors.forename: Masoud Reza
authors.initials: MR
authors.lastname: Sohrabi
keywords: High-Resolution Melting (HRM); mutation analysis; metastatic colorectal cancer (mCRC)