Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 11-19 |
| Number of pages | 9 |
| Journal | Journal of Immunological Methods |
| Volume | 206 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Aug 7 1997 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Immunology
Cite this
Detection of colorectal cancer k-ras mutations using a simplified oligonucleotide ligation assay. / Rothschild, Cynthia B.; Brewer, Catherine S.; Loggie, Brian W.; Beard, G. Alan; Triscott, Mark X.
In: Journal of Immunological Methods, Vol. 206, No. 1-2, 07.08.1997, p. 11-19.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of colorectal cancer k-ras mutations using a simplified oligonucleotide ligation assay
AU - Rothschild, Cynthia B.
AU - Brewer, Catherine S.
AU - Loggie, Brian W.
AU - Beard, G. Alan
AU - Triscott, Mark X.
PY - 1997/8/7
Y1 - 1997/8/7
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0030809713&partnerID=8YFLogxK
U2 - 10.1016/S0022-1759(97)00078-1
DO - 10.1016/S0022-1759(97)00078-1
M3 - Article
C2 - 9328563
AN - SCOPUS:0030809713
VL - 206
SP - 11
EP - 19
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -