title: [Detection of K-ras mutations in colon cancer by PHA-PHFA (preferential homoduplex formation assay)].
abstract: 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.
authors.affiliation: Department of Molecular Diagnostics, Tohoku University, School of Medicine, Sendai.
authors.forename: T
authors.initials: T
authors.lastname: Funato
authors.forename: Z
authors.initials: Z
authors.lastname: Zeng
authors.forename: J
authors.initials: J
authors.lastname: Satoh
authors.forename: J
authors.initials: J
authors.lastname: Fujiwara
authors.forename: M
authors.initials: M
authors.lastname: Kaku
authors.forename: T
authors.initials: T
authors.lastname: Sasaki
mesh_terms: D015179:Colorectal Neoplasms / Q000235:genetics*; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*