GSTT1 and GSTM1 gene copy number analysis in paraffin-embedded tissue using quantitative real-time PCR

被引：12

作者：

Bediaga, Naiara G. ^{[1
]}

Alfonso-Sanchez, Miguel A. ^{[1
]}

de Renobales, Mertxe ^{[1
]}

Rocandio, Ana M. ^{[2
]}

Arroyo, Marta ^{[2
]}

de Pancorbo, Marian M. ^{[1
]}

机构：

[1] Univ Basque Country, Serv Genom Banco ADN, Fac Farm, Dept Bioquim & Biol Mol, Vitoria 01006, Spain

[2] Univ Basque Country, Dept Nutr & Bromatol, Fac Farm, Vitoria 01006, Spain

来源：

ANALYTICAL BIOCHEMISTRY | 2008年 / 378卷 / 02期

关键词：

D O I：

10.1016/j.ab.2008.04.010

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

GSTT1 and GSTM1 genes possess an inherited deletion associated with a lack of enzyme activity. The heterozygous condition of this deletion is difficult to determine in low-quality DNA with existing PCR protocols. We designed and validated a multiplex real-time PCR assay by adapting the Delta Delta Ct relative quantification method for the analysis of GSTY1 and GSTM1 markers to accurately differentiate the three genotypes (*1/1, *1/0, and *0/0) in degraded DNA from formalin-fixed paraffin-embedded tissue. Gene copy number values obtained provide for unambiguous homozygous and heterozygous differentiation. The efficacy shown by the PCR assay endorses its usefulness for complete genotyping of glutathione S-transferases in archival tissues. (C) 2008 Elsevier Inc. All rights reserved.

引用

页码：221 / 223

页数：3

共 10 条

[1] Possible gene dosage effect of glutathione-S-transferases on atopic asthma: Using real-time PCR for quantification of GSTM1 and GSTT1 gene copy numbers [J].

Brasch-Andersen, C ;

Christiansen, L ;

Tan, QH ;

Haagerup, A ;

Vestbo, J ;

Kruse, TA .

HUMAN MUTATION, 2004, 24 (03) :208-214

[2]

Brockmoller J, 1996, CANCER RES, V56, P3915

[3] Quantitative real-time PCR for gene dosage determinations in microdeletion genotypes [J].

Covault, J ;

Abreu, C ;

Kranzler, H ;

Oncken, C .

BIOTECHNIQUES, 2003, 35 (03) :594-+

[4] Trimodal GSTT1 and GSTM1 genotyping assay by real-time PCR [J].

Girault, I ;

Lidereau, R ;

Bièche, I .

INTERNATIONAL JOURNAL OF BIOLOGICAL MARKERS, 2005, 20 (02) :81-86

[5] Real-time PCR analysis of DNA and RNA extracted from formalin-fixed and paraffin-embedded biopsies [J].

Lehmann, U ;

Kreipe, H .

METHODS, 2001, 25 (04) :409-418

[6] Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method [J].

Livak, KJ ;

Schmittgen, TD .

METHODS, 2001, 25 (04) :402-408

[7] GSTM1, GSTT1, and GSTP1 polymorphisms and risk of advanced colorectal adenoma [J].

Moore, LE ;

Huang, WY ;

Chatterjee, N ;

Gunter, M ;

Chanock, S ;

Yeager, M ;

Welch, B ;

Pinsky, P ;

Weissfeld, J ;

Hayes, RB .

CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION, 2005, 14 (07) :1823-1827

[8] Association of homozygous wild-type glutathione S-transferase M1 genotype with increased breast cancer risk [J].

Roodi, N ;

Dupont, WD ;

Moore, JH ;

Parl, FF .

CANCER RESEARCH, 2004, 64 (04) :1233-1236

[9] Characterization of the glutathione S-transferase GSTT1 deletion:: discrimination of all genotypes by polymerase chain reaction indicates a trimodular genotype-phenotype correlation [J].

Sprenger, R ;

Schlagenhaufer, R ;

Kerb, R ;

Bruhn, C ;

Brockmöller, J ;

Roots, I ;

Brinkmann, U .

PHARMACOGENETICS, 2000, 10 (06) :557-565

[10]

Wiebel FA, 1999, PHARMACOGENETICS, V9, P251

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