CYP2D6 Haplotype Determination Using Long Range Allele-Specific Amplification: Resolution of a Complex Genotype and a Discordant Genotype Involving the CYP2D6*59 Allele

The Journal of Molecular Diagnostics : JMD
Andrea GaedigkJ Steven Leeder

Abstract

Cytochrome P450 (CYP) 2D6, a major contributor to the metabolism and bioactivation of many clinically used drugs, is encoded by a complex, highly polymorphic gene locus. To aid in the characterization of CYP2D6 allelic variation, we developed allele-specific long-range PCR (ASXL-PCR) to amplify only the allele of interest for further characterization by PCR. This development was achieved utilizing single-nucleotide polymorphisms in the upstream region of CYP2D6 and a universal CYP2D6-specific reverse primer. This approach was assessed and optimized on samples with known genotypes. The application of ASXL-PCR clarified a case with a complex genotype (CYP2D6*2x2/*4N+*4) by amplifying the duplicated gene units separately for subsequent analysis. Furthermore, ASXL-PCR and subsequent sequence analysis also resolved genotype discord in a mother/daughter relationship by revealing the presence of the CYP2D6*59 allelic variant in both individuals. Finally, we demonstrated that the 2939G>A single-nucleotide polymorphism present on CYP2D6*59 interfered with the TaqMan genotype assay that detected 2850C>T, causing false genotype assignments. Assay interference was resolved using an alternative TaqMan genotype assay currently available as a...Continue Reading

References

Nov 18, 2003·Naunyn-Schmiedeberg's Archives of Pharmacology·Ulrich M ZangerMichel Eichelbaum
Jan 18, 2006·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Andrea GaedigkJ Steven Leeder
Sep 27, 2006·Pharmacogenetics and Genomics·Claudia ToscanoUlrich M Zanger
Jan 19, 2008·European Journal of Clinical Pharmacology·Andrea Gaedigk, Christa Coetsee
Aug 13, 2008·Analytical and Bioanalytical Chemistry·Ulrich M ZangerMatthias Schwab
Sep 18, 2008·European Journal of Clinical Pharmacology·Andrea GaedigkUwe Fuhr
Dec 2, 2009·The Journal of Molecular Diagnostics : JMD·Ralph StadhoudersMartin Schutten
May 18, 2010·European Journal of Clinical Pharmacology·Andrea GaedigkSusan Corey
Jan 1, 2010·Frontiers in Pharmacology·Andrea GaedigkJ Steven Leeder
Nov 25, 2011·Pharmacogenomics·Andrea GaedigkJ Steven Leeder
Dec 22, 2011·Drug Metabolism and Pharmacokinetics·Lay Kek Teh, Leif Bertilsson
Oct 25, 2013·International Review of Psychiatry·Andrea Gaedigk
Jan 15, 2015·Psychiatry Journal·Kathryn R GardnerJay Lombard
Mar 20, 2015·Scientific Reports·Andrea GaedigkJ Steven Leeder

❮ Previous
Next ❯

Citations

Jan 23, 2016·Frontiers in Pharmacology·Amanda K RiffelAndrea Gaedigk
Jul 19, 2018·Pharmacogenomics·Charity Nofziger, Markus Paulmichl
Nov 26, 2015·Human Mutation·Wanqiong QiaoStuart A Scott
Sep 6, 2017·Omics : a Journal of Integrative Biology·Fernando de AndrésAdrián LLerena
Sep 24, 2019·Clinical Pharmacology and Therapeutics·Charity NofzigerAndrea Gaedigk
Feb 8, 2019·Pharmacogenomics·Wanqiong QiaoStuart A Scott
Sep 14, 2018·NPJ Genomic Medicine·Gouri MukerjeeRuslan Dorfman
Feb 26, 2021·The Journal of Molecular Diagnostics : JMD·Amy J TurnerAndrea Gaedigk
Oct 31, 2021·Clinical Pharmacology and Therapeutics·Jean C DinhAndrea Gaedigk

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.