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High sensitivity of reverse-hybridization methodology in the detection of KRAS mutations from formalin-fixed paraffin-embedded colorectal cancer samples

De Miglio, Maria Rosaria and Mura, Antonica and Uras, Maria Gabriela and Manca, Alessandra and Contini, Marcella and Murgia, Luciano and Zinellu, Angelo and Sotgia, Salvatore and Carru, Ciriaco and Massarelli, Giovannino and Cossu Rocca, Paolo Alessandro (2010) High sensitivity of reverse-hybridization methodology in the detection of KRAS mutations from formalin-fixed paraffin-embedded colorectal cancer samples. Diagnostic Molecular Pathology, Vol. 19 (4), p. 201-208. ISSN 1052-9551. Article.

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DOI: 10.1097/PDM.0b013e3181db67d5

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Abstract

Colorectal cancer is ranked the third most common cancer worldwide in terms of incidence and the second in terms of mortality. Recent advances in therapeutic approaches to colorectal cancer have identified a potential role of antiepidermal growth factor receptor (EGFR) targeted therapies as adjuvant treatment in advanced disease. New evidences showed that patients harboring KRAS mutations on codons 12 and 13 are not responsive to anti-EGFR monoclonal antibodies. Therefore, new mutational screening tools have been proposed to select patients who will benefit from anti-EGFR targeted therapy, reducing inappropriate, expensive treatments and unwarranted side effects. We evaluated the performance of a reverse-hybridization-based assay in the identification of the most frequent KRAS mutations on a series of 50 formalin-fixed, paraffin-embedded, advanced colorectal cancer specimens, in comparison with the direct gene sequencing technique. Thirtytwo of the 50 cases (64%) showed KRAS single point mutations by reverse-hybridization technique. In particular, 93.8% of the mutations were reported on codon 12, whereas 6.2% of the mutations were reported on codon 13. Direct gene sequencing showed KRAS mutations on 28 of the 50 cases (56%) with 96.4% of the mutations on codon 12 and 3.6% on codon 13. Concordance between the assays was observed in 92% of the cases. Both reverse hybridization and gene sequencing methods have been shown to be suitable tests in detecting KRAS mutations from formalin-fixed, paraffin-embedded tumor specimens. In our experience, reverse-hybridization technique has been shown to be an effective and more sensitive assay for the identification of the most common KRAS mutations.

Item Type:Article
ID Code:6309
Status:Published
Refereed:Yes
Uncontrolled Keywords:KRAS, EGFR, diagnostic test, colorectal cancer, sequencing, reverse-hybridization assay
Subjects:Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Area 06 - Scienze mediche > MED/08 Anatomia patologica
Area 06 - Scienze mediche > MED/04 Patologia generale
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
001 Università di Sassari > 03 Istituti > Anatomia patologica
Publisher:Lippincott Williams & Wilkins
ISSN:1052-9551
Copyright Holders:© 2010 by Lippincott Williams & Wilkins
Deposited On:25 Jul 2011 13:51

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