[Detection of K-ras mutations in colon cancer by PHA-PHFA (preferential homoduplex formation assay)].
The mutations of K-ras gene have been demonstrated at frequencies of about 40% in human colorectal cancer. We applied a developed PCR-preferential homoduplex formation assay (PCR-PHFA) to detect a point mutation of K-ras gene in the surgical specimens from thirty patients with colorectal cancer. This method is based on the strand competition during hybridization between a double labeled amplicon, prepared from biotin and DNP labeled primers, and an unlabeled amplicon. The procedure of this method is simple and speedy, and suitable to detect mutations in a small number of samples. By using this method, the mutations were found in 37% (11/30) and confirmed by sequencing analysis. The results suggest that the PCR-PHFA system may be useful for detecting low frequent mutation of K-ras gene even in the case of an early cancer.
Funato
T
T
Department of Molecular Diagnostics, Tohoku University, School of Medicine, Sendai.
Zeng
Z
Z
Satoh
J
J
Fujiwara
J
J
Kaku
M
M
Sasaki
T
T
D015179:Colorectal Neoplasms / Q000235:genetics*; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*