Assay design and optimization of mutant-enriched PCR based method for detection of K-ras gene mutations in pancreatic carcinoma

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...

Full description

Saved in:
Bibliographic Details
Published inNeoplasma Vol. 53; no. 5; p. 363
Main Authors Ugorcáková, J, Hlavatý, T, Babál, P, Janega, P, Bukovská, G, Halgasová, N, Hlavatá, A, Timko, J, Duris, I
Format Journal Article
LanguageEnglish
Published Slovakia 2006
Subjects
Adenocarcinoma - genetics
Base Sequence
Colorectal Neoplasms - genetics
DNA Primers
Genes, ras
Humans
Molecular Sequence Data
Mutation
Pancreatic Neoplasms - diagnosis
Pancreatic Neoplasms - genetics
Polymerase Chain Reaction - economics
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Time Factors
Online AccessGet more information

Cover

More Information
Summary: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.
ISSN:0028-2685