Rapid Detection of CYP1A1, CYP2D6, and NAT Variants by Multiplex Polymerase Chain Reaction and Allele-Specific Oligonucleotide Assay

doi:10.1006/abio.1999.4293

Analytical Biochemistry

Volume 275, Issue 1, 1 November 1999, Pages 84-92

https://doi.org/10.1006/abio.1999.4293 Get rights and content

Abstract

Drugs and carcinogens are excreted from the body after metabolic conversion involving enzymes mediating oxidative metabolism and conjugation. Many of the corresponding genes exhibit functional polymorphisms that contribute to individual cancer susceptibility. To increase the efficiency and to facilitate genotyping, we developed a combined approach (PCR–ASO) which includes multiplex PCR and allele-specific oligonucleotide (ASO) hybridization. PCR primer pairs were used to amplify the following alleles/variants: CYP1A1*1, *2A, *2B; CYP2D6*3, *4; NAT1*4, *3, *10, *11, *14, *15; and NAT2*4, *5A, *5B, *5C, *6A, *7B. The products were dot-blotted and polymorphisms were detected by hybridization with ASO probes for both wild-type and variant sites in parallel. This approach was validated by genotyping DNA samples from a French–Canadian population that was previously analyzed by PCR–RFLP. The variants frequencies were compared with the data on other populations available in the literature. The PCR–ASO assay appears to be simple, efficient, and cost-effective, particularly if a large number of samples are to be screened for several DNA variants. This approach has potential for automation with microplates and robotic workstations for high throughput.

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¹: To whom correspondence should be addressed at Centre de recherche, Hôpital Sainte-Justine, 3175 Côte-Sainte Catherine, Montréal, Québec, H3T 1C5 Canada. Fax: (514) 345-4731. E-mail: [email protected].

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Regular ArticleRapid Detection of CYP1A1, CYP2D6, and NAT Variants by Multiplex Polymerase Chain Reaction and Allele-Specific Oligonucleotide Assay

Abstract

FEBS Lett.

Toxicol. Lett.

Leuk. Res.

Anal. Biochem.

Blood

Am. J. Hum. Genet.

Genetic linkage of lung cancer-associated Msp1 polymorphism with amino acid replacement in the heme binding region of the human cytochrome P4501A1 gene

J. Biochem.

C4887A polymorphism in exon 7 of human CYP1A1: Population frequency, mutation linkages, and impact on lung cancer susceptibility

Cancer Res.

Cytochrome P450 2D6 variants in a Caucasian population: Allele frequencies and phenotypic consequences

Am. J. Hum. Genet.

Arylamine N-acetyltransferase (NAT2) mutations and their allelic linkage in unrelated Caucasian individuals: Correlation with phenotypic activity

Am. J. Hum. Genet.

Ethnic distribution of slow acetylator mutations in the polymorphic N-acetyltransferase (NAT2) gene

Pharmacogenetics

Nomenclature for N-acetyltransferase

Pharmacogenetics

Molecular epidemiology: Insights into cancer susceptibility, risk assessment, and prevention

J. Natl. Cancer Inst.

Racial differences in restriction fragment length polymorphisms and messenger RNA inducibility of the human CYP1A1 gene

Cancer Epidemiol. Biomarkers Prev.

Homozygous rapid arylamine N-acetyltransferase (NAT2) genotype as a susceptibility factor in lung cancer

Cancer Res.

Polycyclic aromatic hydrocarbon–DNA adducts in human lung and cancer susceptibility genes

Cancer Res.

Debrisoquine hydroxylase gene polymorphism and susceptibility to Parkinson's disease

Lancet

Combined analysis of Inherited polymorphisms in arylamine N-acetyltransferase 2, glutathione S-transferases M1, and T1, microsomal epoxide hydrolase, and cytochrome P450 enzymes as modulators of bladder cancer risk

Cancer Res.

The human debrisoquine 4-hydroxylase (CYP2D) locus: Sequence and identification of the polymorphic CYP2D6 gene, a related gene, and a pseudogene

Am. J. Hum. Genet.

Regular Article
Rapid Detection of CYP1A1, CYP2D6, and NAT Variants by Multiplex Polymerase Chain Reaction and Allele-Specific Oligonucleotide Assay