title: Detection of RET proto-oncogene codon 634 mutations using mass spectrometry.
abstract: Mutations located in the RET proto-oncogene at codon 634 associated with multiple endocrine neoplasia type 2A and medullary thyroid carcinoma are detected by low-resolution and high-resolution mass spectrometry schemes not requiring labeling or electrophoretic separation of diagnostic products. The former requires measurement by matrix-assisted laser desorption ionization time-of-flight mass spectrometry of 21- to 27-mer oligonucleotides generated by a primer oligo base extension reaction. The latter is based upon direct measurement of artificial products which include the mutation site using matrix-assisted laser desorption ionization Fourier transform mass spectrometry. In this feasibility study a synthetic 25-mer representing the wildtype allele (7660.3 Da) was easily distinguished from G to A (7644.3 Da) and G to T (7635.3 Da) mutant alleles; the mutant alleles, which differed in mass by only 9.0 Da, were easily resolved when analyzed as a mixture. The results of both detection schemes were highly accurate and reliable, indicating mass spectrometry to be a high-quality alternative for future DNA diagnostics performed in clinical laboratories and genetic profiling studies.
authors.affiliation: Sequenom Instruments GmbH, Hamburg, Germany.
authors.forename: D P
authors.initials: DP
authors.lastname: Little
authors.forename: A
authors.initials: A
authors.lastname: Braun
authors.forename: B
authors.initials: B
authors.lastname: Darnhofer-Demar
authors.forename: A
authors.initials: A
authors.lastname: Frilling
authors.forename: Y
authors.initials: Y
authors.lastname: Li
authors.forename: R T
authors.initials: RT
authors.lastname: McIver
authors.forename: H
authors.initials: H
authors.lastname: Köster
mesh_terms: D001483:Base Sequence; D018276:Carcinoma, Medullary / Q000235:genetics; D029721:Drosophila Proteins; D006580:Genetic Carrier Screening; D006720:Homozygote; D006801:Humans; D018813:Multiple Endocrine Neoplasia Type 2a / Q000235:genetics*; D015345:Oligonucleotide Probes; D017354:Point Mutation; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051096:Proto-Oncogene Proteins c-ret; D011519:Proto-Oncogenes; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D013964:Thyroid Neoplasms / Q000235:genetics*
chemical_list: D029721:Drosophila Proteins; C000715470:MAS1 protein, human; D015345:Oligonucleotide Probes; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila