Real-time genotyping of cytochrome P4501A1 A4889G and T6235C polymorphisms

Harth, Volker; Brüning, Thomas; Abel, Josef; Koch, Brigitte; Berg, Inge E.T. van den; Sachinidis, Agapios; Bolt, H. M.; Vetter, Hans; Y, Ko

doi:10.1006/mcpr.2001.0349

Cited by 24 publications

(11 citation statements)

References 12 publications

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“…The NQO1*2 was performed following the method of Harth et al [39] that briefly consists of a real-time PCR amplification of a 300 bp region on exon 6 of the gene using the primers NQO1F and NQO1R and the corresponding hybridization probes.…”

Section: Polymorphisms Genotypingmentioning

confidence: 99%

CASP8 D302H polymorphism delays the age of onset of breast cancer in BRCA1 and BRCA2 carriers

Suela

Cardeñosa²,

Saéz-González³

et al. 2009

Breast Cancer Res Treat

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The polymorphic genetic differences among individuals may modify the high risk for breast cancer (BC) and/or ovarian cancer (OC) susceptibility conferred by BRCA1 and BRCA2 mutations. In the present study we investigate the relevance of RAD51 -135C > G, TP53 R72P, NQO1*2 and CASP8 D302H polymorphisms as potential modifiers of BC and/or OC susceptibility conferred by these mutations. The study group encompasses 390 BRCA1/BRCA2 mutation carriers (182 affected with BC and/or OC and 208 unaffected) of 131 unrelated families studied in the Program of Genetic Counselling on Cancer of Valencia Community. The polymorphisms were detected in genomic DNA by ASRA method or real time PCR using fluorescently labeled probes. We found similar incidence of RAD51 -135C > G, TP53 R72P and NQO1*2 polymorphisms among affected and unaffected individuals considering BRCA1/BRCA2 mutations together and separately. However, the CASP8 D302H polymorphism was strongly associated with the absence of BC [OR = 3.41 (95% CI 1.33-8.78, P = 0.01)]. In fact, in the females with CASP8 D302H polymorphism the BC appeared at a median age of 58 in opposition to the 47 years observed for the wild type subjects (P = 0.03). Furthermore, the CASP8 D302H positive females showed a 50% probability of being free of BC by the age of 78 versus the 2% of the CASP8 negative ones. Our results support that the presence of the CASP8 D302H polymorphism diminishes the high risk of BC conferred by BRCA1 and BRCA2 mutations, making possible that some of the carriers could escape from suffering BC along their life span.

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Section: Polymorphisms Genotypingmentioning

confidence: 99%

CASP8 D302H polymorphism delays the age of onset of breast cancer in BRCA1 and BRCA2 carriers

Suela

Cardeñosa²,

Saéz-González³

et al. 2009

Breast Cancer Res Treat

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“…We and others have used this method for identification of polymorphisms of glutathione S-transferases (GST), different CYP450 isoenzymes or a1-antitrypsin; it allows the analysis of 30 samples within 60 min (Bru¨ning et al 1999;Ortiz-Pallardo et al 2000;Ko et al 2000Ko et al , 2001Harth et al 2001;Fronhoffs et al 2002;Molde et al 2002;Rodriguez et al 2002;Ballerini et al 2003;Wenghoefer et al 2003). In these cases, the DNA used was derived either from whole blood or from paraffinembedded archival tissue specimens.…”

Section: Introductionmentioning

confidence: 99%

Reliability of non-invasively acquired human genomic DNA as a substrate for real-time PCR-assisted analysis of genetic polymorphisms

et al. 2004

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Molecular epidemiological studies require high numbers of participants. The combination of an non-invasive access to human DNA with a rapid genotyping analysis, e.g. by use of LightCycler assisted real-time polymerase chain reaction (PCR), can be helpful in conducting such trials. The aim of our study was to define, for the first time, the use of LightCycler technology in analysis of non-invasively derived DNA. DNA extracted from blood, mouthwash and buccal cytobrush samples from 100 volunteers was analyzed for the genotypes of cytochrome P450 CYP1B1, and glutathione S-transferases GSTT1, GSTM1 and GSTP1. The median amounts of DNA isolated from blood, mouthwash and buccal cytobrush samples were 95, 11 and 8 microg, respectively. While genotyping for CYP1B1 codon 432 polymorphism and GSTP1 codon 105 polymorphism resulted in a complete correspondence for all three modes of sampling, the identification of individuals with null-genotype for GSTT1 or GSTM1 failed in some cases due to atypical courses of the corresponding melting curves, leading to high false-positive rates in the group of non-invasively derived samples. Thus, the results presented here call for caution in using LightCycler assisted real-time PCR in non-invasively collected samples, at least when appropriate control strategies are not implemented.

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“…8,[17][18][19] First, RFLP can in some instances result in significant false positive rates as a result of incomplete restriction enzyme digestion or the presence of other mutations close to the mutation of interest. 20,21 Second, fluorescence-based genotyping assays are more amenable to high-throughput screening, as they do not require extensive postamplification manipulation. Commonly used fluorescence-based PCR techniques for SNP detection include the use of either the nonspecific DNA intercalating dye SYBR Green I or an allele-specific fluorogenic probe (ie, Taqman).…”

Section: Resultsmentioning

confidence: 99%

Detection of MDR1 single nucleotide polymorphisms C3435T and G2677T using real-time polymerase chain reaction: MDR1 single nucleotide polymorphism genotyping assay

Song

Shen

Meibohm

et al. 2002

AAPS J

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The objective of this study was to develop a real-time polymerase chain reaction (PCR) method to detect MDR1 (human multidrug resistance gene) single nucleotide polymorphisms (SNPs) C3435T and G2677T. C3435T and G2677T are l inked to MDR1*2 , which is associated with enhanced efflux activity in vitro. Using the Smart Cycler, an allele-specific real-time PCR-based genotyping method was developed to detect C3435T and G2677T. The MDR1 genotype of human genomic DNA templates was determined by direct DNA sequencing. PCR reactions for genotyping C3435T and G2677T by using allele-specific primers were conducted in separate tubes. An additional nucleotide mismatch at the third position from the 3' end of each allele-specific primer was used to abrogate nonspecific PCR amplification. The fluorescence emitted by SYBR Green I was monitored to detect formation of specific PCR products. PCR growth curves exceeding the threshold cycle were considered positive. Fluorescence melt-curve analysis was used to corroborate results from PCR growth curves. Using PCR growth curves, our assay accurately determined hetero-and homozygosity for C3435T and G2677T. Genotype assignments based on PCR growth curve, melt-curve analysis, agarose gel electrophoresis, and direct DNA sequencing results of PCR products were in perfect agreement. We have developed a rapid MDR1 genotyping method that can be used to assess the contribution of MDR1*2 to pharmacokinetic and pharmacodynamic variability of Pglycoprotein substrates.

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Real-time genotyping of cytochrome P4501A1 A4889G and T6235C polymorphisms

Cited by 24 publications

References 12 publications

CASP8 D302H polymorphism delays the age of onset of breast cancer in BRCA1 and BRCA2 carriers

CASP8 D302H polymorphism delays the age of onset of breast cancer in BRCA1 and BRCA2 carriers

Reliability of non-invasively acquired human genomic DNA as a substrate for real-time PCR-assisted analysis of genetic polymorphisms

Detection of MDR1 single nucleotide polymorphisms C3435T and G2677T using real-time polymerase chain reaction: MDR1 single nucleotide polymorphism genotyping assay

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