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A single, multiplex analysis for all relevant activating NRAS gene mutations using heteroduplex generators
Author(s) -
Belli Carolina,
Bowen Derrick J.,
Brasi Carlos De,
Larripa Irene
Publication year - 2004
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.2004.05079.x
Subject(s) - heteroduplex , multiplex , exon , multiplex polymerase chain reaction , genetics , neuroblastoma ras viral oncogene homolog , biology , mutation , microbiology and biotechnology , gene , mutation testing , polymerase chain reaction , kras
Summary We describe a multiplex polymerase chain reaction (PCR)‐based test that detected all relevant NRAS activating mutations using a single PCR followed directly by electrophoresis. The test uses a Universal Heteroduplex Generator (UHG) to detect exon‐2 (codon 61) NRAS mutations in multiplex with an UHG for exon‐1 (codons 12 and 13). The method differentiated all 19 relevant mutations in these exons and showed a mutation independent sensitivity of approximately 6%. The sensitive, specific detection of all NRAS activating mutations using this single rapid test represents a minimum workload and could be applied readily for large‐scale screening and for routine analysis.