Identification of CYP2D6 alleles by single nucleotide polymorphism analysis using pyrosequencing
2003 (English)In: European Journal of Clinical Pharmacology, ISSN 0031-6970, Vol. 59, no 7, 521-526 p.Article in journal (Refereed) Published
OBJECTIVE: To develop a single nucleotide polymorphism (SNP) analysis for identification of cytochrome P450 (CYP)2D6 alleles by pyrosequencing.
METHODS: Swedish blood donors ( n=282) were typed for a partial CYP2D6 genotype comprising the alleles *1 (wild type), *2 (2850C>T), *3 (2549A>del), *4 (1846G>A) and *6 (1707T>del) using polymerase chain reaction (PCR) and pyrosequencing analysis. CYP2D6*5 (CYP2D6 deleted) was identified using an established long multiplex PCR method. Pyrosequencing is a sequencing-by-synthesis method in which a cascade of enzymatic reactions yields detectable light, which is proportional to the incorporated nucleotides. One feature of typing SNPs by pyrosequencing is that each allelic variant will give a unique sequence. These variants can be readily distinguished by pattern recognition software.
RESULTS: Of 281 individuals analysed, 24 (8.5%) were found to be poor metabolisers with two non-functional alleles. This is in the range of 7-10%, previously reported for Caucasians. A total of 126 individuals (45%) had one functional and one non-functional allele and 131 individuals (47%) had two functional alleles.
CONCLUSION: Pyrosequencing was found to be a fast and efficient tool for genotyping. The method is robust, reliable, rapid and has high throughput.
Place, publisher, year, edition, pages
2003. Vol. 59, no 7, 521-526 p.
CYP2D6 - Pyrosequencing - Pharmacogenetics
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:liu:diva-17932DOI: 10.1007/s00228-003-0654-7PubMedID: 13680033OAI: oai:DiVA.org:liu-17932DiVA: diva2:213005