TY - JOUR
T1 - Molecular methods for the detection of mutations
AU - Monteiro, C.
AU - Marcelino, L. A.
AU - Conde, A. R.
AU - Saraiva, C.
AU - Giphart-Gassler, M.
AU - De Nooij-van Dalen, A. G.
AU - Van Buuren-van Seggelen, V.
AU - Van Der Keur, M.
AU - May, C. A.
AU - Cole, J.
AU - Lehmann, A. R.
AU - Steinsgrimsdottir, H.
AU - Beare, D.
AU - Capulas, E.
AU - Armour, J. A L
PY - 2000
Y1 - 2000
N2 - We report the results of a collaborative study aimed at developing reliable, direct assays for mutation in human cells. The project used common lymphoblastoid cell lines, both with and without mutagen treatment, as a shared resource to validate the development of new molecular methods for the detection of low-level mutations in the presence of a large excess of normal alleles. As the 'gold standard,' hprt mutation frequencies were also measured on the same samples. The methods under development included i) the restriction site mutation (RSM) assay, in which mutations lead to the destruction of a restriction site; ii) minisatellite length-change mutation, in which mutations lead to alleles containing new numbers of tandem repeat units; iii) loss of heterozygosity for HLA epitopes, in which antibodies can be used to direct selection for mutant cells; iv) multiple fluorescence-based long linker arm nucleotides assay (mf-LLA) technology, for the detection of substitutional mutations; v) detection of alterations in the TP53 locus using a (CA) array as the target for the screening; and vi) PCR analysis of lymphocytes for the presence of the BCL2 t(14:18) translocation. The relative merits of these molecular methods are discussed, and a comparison made with more 'traditional' methods. (C) 2000 Wiley-Liss, Inc.
AB - We report the results of a collaborative study aimed at developing reliable, direct assays for mutation in human cells. The project used common lymphoblastoid cell lines, both with and without mutagen treatment, as a shared resource to validate the development of new molecular methods for the detection of low-level mutations in the presence of a large excess of normal alleles. As the 'gold standard,' hprt mutation frequencies were also measured on the same samples. The methods under development included i) the restriction site mutation (RSM) assay, in which mutations lead to the destruction of a restriction site; ii) minisatellite length-change mutation, in which mutations lead to alleles containing new numbers of tandem repeat units; iii) loss of heterozygosity for HLA epitopes, in which antibodies can be used to direct selection for mutant cells; iv) multiple fluorescence-based long linker arm nucleotides assay (mf-LLA) technology, for the detection of substitutional mutations; v) detection of alterations in the TP53 locus using a (CA) array as the target for the screening; and vi) PCR analysis of lymphocytes for the presence of the BCL2 t(14:18) translocation. The relative merits of these molecular methods are discussed, and a comparison made with more 'traditional' methods. (C) 2000 Wiley-Liss, Inc.
KW - BCL2 (AR Lehmann)
KW - HLA locus
KW - Hprt
KW - Human population monitoring
KW - Instability
KW - Mf-LLA
KW - Minisatellite
KW - Minisatellites
KW - Mutagenesis (J Cole) restriction site mutations
KW - Small-pool PCR (CA May) p53
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U2 - 10.1002/1520-6866(2000)20:6<357::AID-TCM5>3.0.CO;2-G
DO - 10.1002/1520-6866(2000)20:6<357::AID-TCM5>3.0.CO;2-G
M3 - Article
C2 - 11074520
AN - SCOPUS:0033679789
SN - 0270-3211
VL - 20
SP - 357
EP - 386
JO - Teratogenesis Carcinogenesis and Mutagenesis
JF - Teratogenesis Carcinogenesis and Mutagenesis
IS - 6
ER -