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A fast, cost-efficient and sensitive approach for KRAS mutation detection using multiplexed primer extension with IP/RP-HPLC separation.
- Source :
-
International journal of cancer [Int J Cancer] 2012 Feb 01; Vol. 130 (3), pp. 567-74. Date of Electronic Publication: 2011 Apr 27. - Publication Year :
- 2012
-
Abstract
- Mutations in the KRAS gene are very important diagnostic and prognostic markers in cancer. Particularly, KRAS mutations at codons 12 and 13 have a high prognostic value for EGFR-directed antibody therapies. Several methods are available to detect the most common mutations, some of them are commercialized. The most frequently used techniques, allele-specific PCR or direct sequencing, are not standardized and often lack sensitivity to detect low amounts of mutated tumor cells in paraffin-embedded tissue-blocks leading to a high number of false-negatives. Here we present a reliable, fast, cost-effective and sensitive approach for KRAS mutation detection that has a high potential for standardized large scale screening. The method is based on multiplexed primer extension reactions coupled to HPLC separation. The highly sensitive assay gives easily interpretable and reproducible results at affordable costs. We describe the method and an application example for diagnosis in early colorectal cancer screening.<br /> (Copyright © 2011 UICC.)
- Subjects :
- Early Detection of Cancer
Humans
Polymerase Chain Reaction
Proto-Oncogene Proteins p21(ras)
Reproducibility of Results
Sensitivity and Specificity
Chromatography, High Pressure Liquid methods
Colorectal Neoplasms diagnosis
Colorectal Neoplasms genetics
DNA Mutational Analysis methods
Mutation
Proto-Oncogene Proteins genetics
ras Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1097-0215
- Volume :
- 130
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- International journal of cancer
- Publication Type :
- Academic Journal
- Accession number :
- 21387306
- Full Text :
- https://doi.org/10.1002/ijc.26040