title: A Tetra-Primer Amplification Refractory System Technique for the Cost-Effective and Novel Genotyping of Eight Single-Nucleotide Polymorphisms of the Catechol-O-Methyltransferase Gene.
abstract: OBJECTIVE
  Catechol-O-methyltransferase (COMT) is an enzyme involved in the degradation of catecholamine neurotransmitters. Due to its role in neurotransmitter flux, multiple COMT variants have been associated with the development of psychiatric disorders. Notably, select single-nucleotide polymorphisms (SNPs) of the COMT gene have been implicated in schizophrenia risk, severity, and treatment response. In recognition of the value of a streamlined genotyping method for COMT SNP detection, this study was designed to develop a simple and economical tetra-primer amplification refractory mutation system (T-ARMS) assay for the concurrent detection of eight COMT SNPs: rs4680, rs737865, rs165599, rs2075507, rs4633, rs4818, rs6269, and rs165774.
  
  
  METHODS
  T-ARMS is a genotyping method that uses polymerase chain reaction (PCR) to amplify a multiplex reaction consisting of two primer pairs. T-ARMS primers are customized to each SNP and designed to generate different-sized allele-specific amplicons. This assay was applied to a total of 39 genomic DNA samples. Genotypic designations across the panel of SNPs were subsequently validated by Sanger sequencing.
  
  
  RESULTS
  T-ARMS reliably and unambiguously detected all three genotypes (homozygous wild type, heterozygous, and homozygous mutant) for each of the eight COMT SNPs.
  
  
  CONCLUSIONS
  Compared to traditional low-throughput methods that require post-PCR modification or high-throughput technologies that require sophisticated equipment, T-ARMS is a cost-effective and efficient assay that can be easily adapted by any standard molecular diagnostics laboratory. This T-ARMS assay provides a practical and robust method for COMT SNP detection.
authors.affiliation: 1 Department of Pharmacology, University of British Columbia , Vancouver, Canada .
authors.forename: Cathy K
authors.initials: CK
authors.lastname: Wang
authors.affiliation: 1 Department of Pharmacology, University of British Columbia , Vancouver, Canada .|2 Department of Psychiatry, University of British Columbia , Vancouver, Canada .
authors.forename: Ana
authors.initials: A
authors.lastname: Aleksic
authors.affiliation: 1 Department of Pharmacology, University of British Columbia , Vancouver, Canada .
authors.forename: Michael S
authors.initials: MS
authors.lastname: Xu
authors.affiliation: 2 Department of Psychiatry, University of British Columbia , Vancouver, Canada .
authors.forename: Ric M
authors.initials: RM
authors.lastname: Procyshyn
authors.affiliation: 3 Department of Medical Genetics, University of British Columbia , Vancouver, Canada .
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Ross
authors.affiliation: 2 Department of Psychiatry, University of British Columbia , Vancouver, Canada .
authors.forename: Fidel
authors.initials: F
authors.lastname: Vila-Rodriguez
authors.affiliation: 2 Department of Psychiatry, University of British Columbia , Vancouver, Canada .
authors.forename: Alfredo
authors.initials: A
authors.lastname: Ramos-Miguel
authors.affiliation: 1 Department of Pharmacology, University of British Columbia , Vancouver, Canada .
authors.forename: Ryan
authors.initials: R
authors.lastname: Yan
authors.affiliation: 2 Department of Psychiatry, University of British Columbia , Vancouver, Canada .
authors.forename: William G
authors.initials: WG
authors.lastname: Honer
authors.affiliation: 1 Department of Pharmacology, University of British Columbia , Vancouver, Canada .
authors.forename: Alasdair M
authors.initials: AM
authors.lastname: Barr
mesh_terms: D002394:Catechol O-Methyltransferase / Q000235:genetics*; D003362:Cost-Benefit Analysis; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D005787:Gene Frequency; D005838:Genotype; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods; D006801:Humans; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012559:Schizophrenia / Q000201:enzymology / Q000235:genetics
chemical_list: D017931:DNA Primers; D004247:DNA; C588010:COMT protein, human; D002394:Catechol O-Methyltransferase