Detection of colorectal cancer K-ras mutations using a simplified oligonucleotide ligation assay
| Autor: | Brian W. Loggie, Catherine S. Brewer, Cynthia B. Rothschild, G. Alan Beard, M X Triscott |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Adenoma
Male Colorectal cancer Immunology Enzyme-Linked Immunosorbent Assay Biology Adenocarcinoma medicine.disease_cause Sensitivity and Specificity Exon medicine Immunology and Allergy Humans Point Mutation Genetic Testing Gene Alleles Mutation Oligonucleotide Point mutation Gene Amplification Assay sensitivity Dipstick medicine.disease Molecular biology Genes ras ras Proteins Colorimetry Female Colorectal Neoplasms Oligonucleotide Probes |
| Zdroj: | Journal of immunological methods. 206(1-2) |
| ISSN: | 0022-1759 |
| Popis: | It has been suggested that some mutations in codons 12 and 13 of the K-ras gene are associated with the progression of colorectal adenomas to carcinomas. The aim of this study was to develop a rapid, colorimetric assay for K-ras point mutations commonly associated with colorectal cancer. K-ras exon 1 was amplified from colorectal tumor DNA and K-ras activating mutations detected using an oligonucleotide ligation assay (OLA) in combination with immunological and colorimetric detection. Using the OLA with oligonucleotides specific to individual K-ras mutations, 6 (of 17 total colorectal adenomas/carcinomas) were found to have K-ras mutations. The assay could detect as little as 10% mutant allele. A simplified OLA designed to test for either the presence (+) or absence (−) of any of the K-ras activating mutations was developed. The assay was further streamlined by use of a dipstick methodology for colorimetric development. If required, assay sensitivity can be increased by the use of the recently described EDNA–ELCA detection system. The simplified (+/−) mutation OLA in combination with a dipstick or EDNA–ELCA detection system provides a rapid, sensitive assay for K-ras point mutations suitable for use as part of the clinical assessment of colorectal cancer. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect