DETECTION OF K-RAS POINT MUTATION IN CODON 12 FOR COLORECTAL CANCER BY THE ENZYME-LINKED MINI-SEQUENCE ASSAY IN COMPARISON WITH THE DOT-BLOT HYBRIDIZATION
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The point mutation of K-ras codon 12 in the paraffin-embedded blocks of colorectal cancer from 19 patients was analyzed with a newly developed detection technique, enzyme linked mini-sequence assay (ELMA). The results were compared with those obtained using conventional dot-blot hybridization (DBH) to examine the consistency between these two techniques and the usefulness of the ELMA method. The results of DBH showed that the point mutation of K-ras was present in 12 of 19 patients (63.2%), while ELMA in the first trial showed it in 14 of 19 patients (73.7%). A comparison of the results of DBH with those of ELMA revealed that the presence of the mutation and its arrangement of base patterns agreed in 14 of the 19 patients (73.7%). Then ELMA was repeated for the five patients in whom the results of the two techniques did not agree. It showed that the second ELMA provided the results consistent with those of DBH in four patients, but not in one patient. In three of these four patients, the color intensity of mutation for the first trial of ELMA was (1+). However, the comparison with DBH showed that the results agreed in 90% of the patients judged as (3+) by the first trial of ELMA. The mutations examined by DBH and ELMA and the reproducibility of ELMA agreed in 18 of 19 cases (94.7%) in accordance with the color intensity of 1(+) which should be judged as a negative mutation. We conclude that these results confirmed that ELMA for paraffin-embedded sections was useful in detecting the point mutation of the K-ras gene.
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