Leukemia  2000 Feb;14(2):312-5 

Rapid and reliable detection of N-ras mutations in acute lymphoblastic leukemia
by melting curve analysis using LightCycler technology.

Nakao M, Janssen JW, Seriu T, Bartram CR.

Institute of Human Genetics, University of Heidelberg, Germany.

We applied a new strategy for the detection of N-ras gene mutations based on
LightCycler technology. We designed two sets of amplimers and internal
hybridization probes representing N-ras codons 12/13 and codon 61, respectively.
Genomic DNAs from 134 childhood acute lymphoblastic leukemia (ALL) patients (83
common ALL, nine pre-pre-B ALL, 19 pre-B ALL, 23 T-ALL) were amplified, followed
by the analysis of the melting temperatures of the PCR products on the
LightCycler. PCR products exhibiting an abnormal melting characteristic were
directly sequenced. Sequence analyses unravelled nucleotide substitutions at
codon 12 in 10 patients, at codon 13 in three, and at codon 61 in one case. The
incidence of N-rasmutations (10%) is compatible with previous reports. The
LightCycler technology facilitates the rapid analysis of other genes exhibiting
hot spot mutations in human malignancies.

PMID: 10673750 [PubMed - indexed for MEDLINE]