title: [Detection of KRAS mutations in tumor cells using biochips].
abstract: Somatic mutations in the KRAS gene are important markers of some types of tumors, for example, pancreatic cancer, and may be useful in early diagnostics. A biochip has been developed which allows determining most frequent mutations in 12, 13 and 61 codons of the KRAS gene. To increase the sensitivity of the method and to make possible the analysis of minor fractions of tumor cells in clinical samples the method of blocking a wild type sequence PCR amplification by LNA-oligonucleotides has been used. The product of LNA-clamp PCR was further hybridized with oligonucleotide probes, immobilized on biochip. Biochip was tested with 42 clinical DNA samples from patients with pancreatic cancer, mostly ductal adenocarcinomas. As reference methods, the RFLP analysis and sequencing were used. The developed approach allows detecting somatic mutations in the KRAS gene if the portion of tumor cells with mutation is at least 1% of whole cell population.
authors.forename: M A
authors.initials: MA
authors.lastname: Emel'ianova
authors.forename: F A
authors.initials: FA
authors.lastname: Amosenko
authors.forename: A V
authors.initials: AV
authors.lastname: Chudinov
authors.forename: S A
authors.initials: SA
authors.lastname: Surzhikov
authors.forename: T P
authors.initials: TP
authors.lastname: Kazubskaia
authors.forename: L N
authors.initials: LN
authors.lastname: Liubchenko
authors.forename: T V
authors.initials: TV
authors.lastname: Nasedkina
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis / Q000235:genetics; D000328:Adult; D000368:Aged; D003062:Codon; D015342:DNA Probes / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D057230:Limit of Detection; D008297:Male; D046228:Microarray Analysis / Q000379:methods*; D008875:Middle Aged; D009154:Mutation; D015345:Oligonucleotide Probes / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000175:diagnosis / Q000235:genetics; D057089:Pathology, Molecular / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins / Q000235:genetics*
chemical_list: D003062:Codon; D015342:DNA Probes; C117307:KRAS protein, human; D015345:Oligonucleotide Probes; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins