Back to Search
Start Over
Detection of catechol-O-methyltransferase Val158Met polymorphism by a simple one-step tetra-primer amplification refractory mutation system-PCR
- Source :
-
Molecular & Cellular Probes . Jun2007, Vol. 21 Issue 3, p202-207. 6p. - Publication Year :
- 2007
-
Abstract
- Abstract: The G→A transition at nucleotide 21881 of the human catechol-O-methyltransferase (COMT) gene represents a functional genetic polymorphism (Val158Met), rendering an enzyme with reduced activity that has been associated with psychiatric disorders and estrogen-related cancers. A new method for the detection of this polymorphism is described, based on the tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR), with a single PCR to discriminate both alleles. Two primers amplify a common amplicon independently of the allele considered. At the same time, two primers are used, differing in the 3′ base. In the Val/Val or Met/Met conditions, amplification occurs both in the general amplicon and in the specific allele; in the Val/Met condition three different amplicons are produced. Direct DNA sequencing of a COMT region containing the G/A polymorphism demonstrates the validity of this tetra-primer ARMS-PCR method. Reevaluation by PCR-RFLP revealed 100% accordance for genotype adscription. Subjects carrying the COMT HH genotype in a Spanish population comprised 28%, and the COMT LL homozygotes amounted to 21%. The described method provides a fast and reliable approach for determining COMT polymorphism that can be useful in large clinical studies using minimal quantity of DNA, avoiding the timely and costly use of restriction enzymes. [Copyright &y& Elsevier]
- Subjects :
- *GENETIC polymorphisms
*GENETIC mutation
*POLYPHENOLS
Subjects
Details
- Language :
- English
- ISSN :
- 08908508
- Volume :
- 21
- Issue :
- 3
- Database :
- Academic Search Index
- Journal :
- Molecular & Cellular Probes
- Publication Type :
- Academic Journal
- Accession number :
- 24381349
- Full Text :
- https://doi.org/10.1016/j.mcp.2006.12.001