uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
Cytochrome P450 genotyping by multiplexed real-time DNA sequencing with Pyrosequencing TM technology
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical pharmacogenomics and osteoporosis.ORCID iD: 0000-0002-6368-2622
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Genetics and Pathology.
Responsible organisation
2002 (English)In: Assay and drug development technologies, ISSN 1540-658X, E-ISSN 1557-8127, Vol. 1, no 1, 49-59 p.Article in journal (Refereed) Published
Abstract [en]

Individual differences in xenobiotic metabolism influence the therapeutic value of many drugs and are of major concern during the development of new drug candidates. A number of polymorphic cytochrome p450 enzymes account for a significant part of this variation. A better understanding of these genetic factors would be of value for drug development, as well as clinical practice. To fulfill the goal of a personalized medicine, methods for simple and accurate assessment of cytochrome p450 genes are required. We report on the development of multiplex assays for genotyping of the cytochrome p450 drug-metabolizing enzymes CYP2D6, CYP2C9, and CYP2C19 with Pyrosequencing technology. Eleven variable positions, representing 12 of the most frequent alleles, were scored: CYP2D6 alleles *2, *3, *4, *6, *7, *8, and *14, CYP2C19 alleles *2, *3, and *4, and CYP2C9 alleles *2 and *3. Four multiplex Pyrosequencing reactions per patient sample were performed to cover these positions, using either simplex or multiplex PCR for amplification of target DNA sequences. Unequivocal genotypes were obtained for all patient samples, and the results were validated by comparing with results obtained using PCR-RFLP. For positions addressed with both methods, the results were in complete agreement. Pyrosequencing technology offers a highly automated, rapid, and accurate method for identification of cytochrome p450 alleles, which is suitable for pharmacogenomic research, as well as for routine assessment of patient genotypes.

Place, publisher, year, edition, pages
2002. Vol. 1, no 1, 49-59 p.
Keyword [en]
genotyping, pyrosequence, CYP2D6, CYP2C9, CYP2C19, pharmacogenetics
National Category
Pharmacology and Toxicology
URN: urn:nbn:se:uu:diva-64509OAI: oai:DiVA.org:uu-64509DiVA: diva2:92420
Available from: 2008-01-18 Created: 2008-01-18 Last updated: 2016-02-19

Open Access in DiVA

No full text

Search in DiVA

By author/editor
Wadelius, MiaAlderborn, Anders
By organisation
Clinical pharmacogenomics and osteoporosisDepartment of Genetics and Pathology
In the same journal
Assay and drug development technologies
Pharmacology and Toxicology

Search outside of DiVA

GoogleGoogle Scholar

Total: 127 hits
ReferencesLink to record
Permanent link

Direct link