Cb. Rothschild et al., DETECTION OF COLORECTAL-CANCER K-RAS MUTATIONS USING A SIMPLIFIED OLIGONUCLEOTIDE LIGATION ASSAY, Journal of immunological methods, 206(1-2), 1997, pp. 11-19
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 t
o carcinomas. The aim of this study was to develop a rapid, colorimetr
ic assay for K-ras point mutations commonly associated with colorectal
cancer. K-ras exon 1 was amplified from colorectal tumor DNA and K-ra
s activating mutations detected using an oligonucleotide ligation assa
y (OLA) in combination with immunological and colorimetric detection.
Using the OLA with oligonucleotides specific to individual K-ms mutati
ons, 6 (of 17 total colorectal adenomas/carcinomas) were found to have
K-ras mutations. The assay could detect as little as 10% mutant allel
e. A simplified OLA designed to test for either the presence (+) or ab
sence (-) of any of the K-rns activating mutations was developed, The
assay was further streamlined by use of a dipstick methodology for col
orimetric 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-E
LCA detection system provides a rapid, sensitive assay for K-ras point
mutations suitable for use as part of the clinical assessment of colo
rectal cancer. (C) 1997 Elsevier Science B.V.