Rapid detection of exon 1 NRAS gene mutations using universal heteroduplex generator technology

摘要: Specific NRAS oncogene missense mutations have been frequently found in some tumors and several hematological diseases, especially those of myeloid origin. There is a wide range PCR-based methods for screening detection exon 1 single-base substitutions. However, there are disadvantages ambiguities associated with these techniques because all them require either separate probes, PCR amplifications, or complicated post-PCR manipulations. This report describes new approach gene at codon 12 13 based on the DNA heteroduplex analysis method. The strategy relies upon differential electrophoretic behavior induced molecules formed by cross-hybridization two PCR-amplified species, sample under synthetic universal generator (UHG). panel mutant variants indicated that this discriminates relevant mutations. sensitivity method was estimated competitive assay where alleles could be detected dilution level to 16 wild-type alleles. UHG technology tested clinical samples previously studied PCR-ASO. methodology highly specific, sensitive, achieves an appreciable reduction workload time it requires one amplification followed polyacrylamide gel electrophoresis standard conditions. We propose may applied as alternative 12-13 easily incorporated into routine assays performed oncology laboratories.