http://www.cnr.it/ontology/cnr/individuo/prodotto/ID129328

Multiplex genotyping of CYP2D6 polymorphisms by MALDI-TOF mass spectrometry. (Abstract/Poster in atti di convegno)

Type

Abstract/Poster in atti di convegno (Classe)
Prodotto della ricerca (Classe)

Label

Multiplex genotyping of CYP2D6 polymorphisms by MALDI-TOF mass spectrometry. (Abstract/Poster in atti di convegno) (literal)

Anno

2008-01-01T00:00:00+01:00 (literal)

Alternative label

Falzoi M., Congeddu E., Mancosu G., Masciocchi J., Mossa A., Mura C., Utzeri S., Rodriguez-Tomè P., Pani L., Saba L. (2008)
Multiplex genotyping of CYP2D6 polymorphisms by MALDI-TOF mass spectrometry.
in 4th Biologie Prospective Santorini Conference, Santorini, Greece
(literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

Falzoi M., Congeddu E., Mancosu G., Masciocchi J., Mossa A., Mura C., Utzeri S., Rodriguez-Tomè P., Pani L., Saba L. (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#descrizioneSinteticaDelProdotto

Cytochrome P450 2D6 (CYP2D6) is a highly polymorphic gene responsible for the metabolism of about 25% of commonly prescribed drugs. The presence of Single Nucleotide Polymorphisms (SNPs) and rearrangements within the gene locus can alter enzyme activity with effects ranging from null to excessive activity. The 67 allelic variants and numerous subvariants are summarized at the Human CYP allele nomenclature committee web site(1). The most widely used genotyping methods for CYP2D6 are often time-consuming and they only detect a restricted number of alleles simultaneously. Conversely, the high-throughput oligonucleotide microarray technologies require special laboratory facilities. Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight (MALDI-TOF) mass spectrometry offers automated and high-throughput analysis of SNPs. Because multiple SNPs can be detected in a single reaction, this technique provides a cost-effective method for SNP genotyping. The basic assay methodology uses multiplex PCR reactions followed by minisequencing single base extension analysis. For CYP2D6, we have designed a new PCR strategy based on the entire gene amplification (6,570 Kb) coupled with multiplex primer extension reactions. This strategy avoids false genotyping results by non-specific co-amplification of the homologous pseudogenes CYP2D7P and CYP2D8P and reduces the number of PCR primers used to select regions containing the targeted SNPs. We have performed five 11-13-plex minisequencing reactions and genotyped 57 SNPs in a random sample of 250 Sardinian individuals using the MassARRAY® Sequenom platform. We have applied PHASE(2) to infer sample haplotypes and a properly developed bioinformatic tool to recognize the corresponding allele variants. The tool uses a database (dbCYP) fed with all information contained in (1) and implements a partial-match strategy to assign the allele name using the tipical set of SNPs that defines each allelic variant. This new platform offers a robust and cheap alternative to many currently available CYP2D6 genotyping approaches. (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni