title: Assay design and optimization of mutant-enriched PCR based method for detection of K-ras gene mutations in pancreatic carcinoma.
abstract: The aim of our work was to develop a fast, reliable and sensitive PCR method to detect K-ras mutations in various clinical samples. There is a need for an unimpeachable method for early diagnosis and/or screening of pancreatic cancer (PC). We optimized and subsequently analyzed four methods based on mutant-enriched PCR for the sensitivity, cost and time expense. Using the selected optimal method we examined codon 12 K- ras mutations in a study population of 59 patients with upper GIT malignancies. Reliability of the genotyping was confirmed by sequencing. By using the best of our modified mutant-enriched PCR methods we achieved sensitivity of 1:1 x 10(5). Further studies are necessary to determine the optimal biological material sampling in PC.
authors.affiliation: Institute of Molecular Biology, Slovak Academy of Sciences, 84551 Bratislava, Slovak Republic. jana.ugorcakova@savba.sk
authors.forename: J
authors.initials: J
authors.lastname: Ugorcáková
authors.forename: T
authors.initials: T
authors.lastname: Hlavatý
authors.forename: P
authors.initials: P
authors.lastname: Babál
authors.forename: P
authors.initials: P
authors.lastname: Janega
authors.forename: G
authors.initials: G
authors.lastname: Bukovská
authors.forename: N
authors.initials: N
authors.lastname: Halgasová
authors.forename: A
authors.initials: A
authors.lastname: Hlavatá
authors.forename: J
authors.initials: J
authors.lastname: Timko
authors.forename: I
authors.initials: I
authors.lastname: Duris
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics*; D001483:Base Sequence; D015179:Colorectal Neoplasms / Q000235:genetics*; D017931:DNA Primers; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010190:Pancreatic Neoplasms / Q000175:diagnosis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D017931:DNA Primers