Rapid and large-scale method to detect K-ras gene mutations in tumor samples
Lopez-Crapez, E.; Chypre, C.; Saavedra, J.; Marchand, J.; Grenier, J.
Clinical Chemistry 43(6 Pt 1): 936-942
1997
ISSN/ISBN: 0009-9147 PMID: 9191543 Document Number: 474201
We have developed a rapid and large-scale method for the detection of K-ras gene mutations in tumors. First, DNA is amplified by an asymmetric PCR; second, the single-strand dinitrophenyl (DNP)-labeled amplified DNA is hybridized specifically to oligonucleotide probes affixed on a tube. Finally, perfectly matched duplexes are easily detected by a monoclonal anti-DNP antibody bearing 125I. The usefulness of this technique is illustrated by analyzing K-ras codon 12 mutations in human colorectal samples. This reliable assay procedure can be applied to the rapid screening of virtually any genetic disease caused by previously described point mutations.