Table of Contents
Sequencing
Volume 2011, Article ID 895709, 7 pages
http://dx.doi.org/10.1155/2011/895709
Research Article

KRAS Codons 12 and 13 Mutation Analysis: A Comparative Study between Direct Sequencing and a New Sensitive Real-Time PCR Assay

1Dipartimento di Scienze Mediche e Biologiche, Università di Udine, 33100 Udine, Italy
2AB ANALITICA s.r.l., 35127 Padova, Italy

Received 29 July 2011; Revised 26 September 2011; Accepted 29 September 2011

Academic Editor: Luis Menéndez-Arias

Copyright © 2011 Stefania Marzinotto et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

KRAS somatic mutations are found in 30–40% of colorectal cancer (CRC). Seven mutations in codons 12 and 13 of KRAS (95% of the observed human mutations) preclude the efficacy of anti-EGFR therapy for the treatment of CRC. Assessment of KRAS mutational status has become a standard procedure in the management of patients with CRC. Technically, KRAS mutation testing can be performed with different methods, characterized by distinct sensitivities and specificities. The present study analyzed KRAS in 182 CRC histological samples by using direct sequencing and a new kit based on a Real-Time Sequence-Specific Primers-PCR technology. The kit allowed to recover as positive 17 samples that were negative or unclear by sequencing, with a recovery rate equal to 13.82%. This study proposes a fast, sensitive, and high-throughput system to identify such seven described mutations of KRAS gene in CRC samples.