NDLI: A commercial real-time PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma.

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A commercial real-time PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma.

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Year: 2010 Volume: 12 Issue: 2

Year: 2010 Volume: 12 Issue: 1

Year: 2009 Volume: 11

Year: 2008 Volume: 10

A commercial real-time PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Angulo, Bárbara García-García, Elena Martínez, Rebeca Suárez-Gauthier, Ana Conde, Esther Hidalgo, Manuel López-Ríos, Fernando
Description	Country affiliation: Spain Author Affiliation: Angulo B ( Hospital Universitario Madrid Sanchinarro, Laboratorio de Dianas Terapeuticas, Calle Oña 10, Madrid, Spain.)
Abstract	KRAS mutation testing has become a standard procedure in the management of patients with carcinomas. The most frequently used method for KRAS testing is direct sequencing of PCR products. The development of commercial real-time quantitative PCR kits offers a useful alternative since they are in theory much more sensitive than direct sequencing and they avoid post- PCR handling. We present our experience as a reference center for the study of KRAS mutations, comparing direct sequencing and the use of a commercial real-time quantitative PCR kit, as well as determining the sensitivity of both procedures in clinical practice. The TheraScreen K-RAS Mutation Kit identified mutations in 75 (44%) of the 170 tumors. Three cases were tested positive using TheraScreen K-RAS Mutation Kit and negative by direct sequencing. We then compared the sensitivity of the kit and that of direct sequencing using 74 mutant tumors. The kit was able to detect the presence of a mutation in a 1% dilution of the total DNA in 13.5% of the tumors and, in 84%, KRAS mutation was identified at a dilution of 5%. Sequencing was able to detect KRAS mutations when the mutant DNA represented 10% of the total DNA in 20/74 (27%) of the tumors. When the mutant DNA represented 30% of the total DNA, sequencing could detect mutations in 56/74 (76%).
File Format	HTM / HTML
ISSN	15251578
e-ISSN	19437811
DOI	10.2353/jmoldx.2010.090139
Journal	The Journal of Molecular Diagnostics
Issue Number	3
Volume Number	12
Language	English
Publisher	Elsevier
Publisher Date	2010-05-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Molecular Biology Colorectal Neoplasms Diagnosis Genetics Polymerase Chain Reaction Proto-oncogene Proteins Sequence Analysis, Dna Ras Proteins Mutation Proto-oncogene Proteins P21(ras) Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Molecular Medicine Pathology and Forensic Medicine

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A Commercial Real-Time PCR Kit Provides Greater Sensitivity than Direct Sequencing to Detect KRAS Mutations

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A commercial real-time PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma.

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A Commercial Real-Time PCR Kit Provides Greater Sensitivity than Direct Sequencing to Detect KRAS Mutations

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A commercial real-time PCR kit provides greater sensitivity than direct sequencing to detect KRAS mutations: a morphology-based approach in colorectal carcinoma.