title: A simultaneous LightCycler detection assay for five genetic polymorphisms influencing drug sensitivity.
abstract: OBJECTIVE
  The routine detection of polymorphisms affecting drug sensitivity in patients before treatment is important in the identification of drug responders or nonresponders, and patients at increased risk of drug toxicity. Here, we present an assay for the simultaneous and rapid genotyping of five polymorphisms influencing drug sensitivity.
  
  
  METHODS
  We used a hybridization probe assay on the LightCycler to detect five single nucleotide polymorphisms (SNPs): INPP1 (973C>A), ADRB2 (R16G and Q27E), HTR2A (102T>C), and mtDNA (1555A>G). Two fluorescent labeled hybridization probes were designed for the simultaneous detection of the five SNPs and detection of the variant alleles was performed by melting curve analysis.
  
  
  RESULTS
  All five SNPs were detected with a single thermocycle protocol within 40 min. The genotypes determined in this assay were identical to those obtained with conventional PCR and restriction fragment length polymorphism analysis.
  
  
  CONCLUSIONS
  To our knowledge, we report here for the first time a method for simultaneous detection of five SNPs, on a single thermocycle protocol by the LightCycler. This method is rapid, highly sensitive, and high-throughput, and is thus suitable for routine clinical use and large-scale epidemiologic studies.
authors.affiliation: Department of Pharmaceutical Sciences, Tohoku University Hospital, 980-8574, Sendai, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Hiratsuka
authors.forename: Kaori
authors.initials: K
authors.lastname: Narahara
authors.forename: Yukinaga
authors.initials: Y
authors.lastname: Kishikawa
authors.forename: Samar
authors.initials: S
authors.lastname: Ismail Hamdy
authors.forename: Naomi
authors.initials: N
authors.lastname: Endo
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Agatsuma
authors.forename: Masaki
authors.initials: M
authors.lastname: Matsuura
authors.forename: Tomoko
authors.initials: T
authors.lastname: Inoue
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Tomioka
authors.forename: Michinao
authors.initials: M
authors.lastname: Mizugaki
mesh_terms: D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000235:genetics*; D004272:DNA, Mitochondrial / Q000235:genetics*; D004351:Drug Resistance; D005838:Genotype; D006801:Humans; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D044402:Receptor, Serotonin, 5-HT2A; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics*; D011985:Receptors, Serotonin / Q000235:genetics*
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004272:DNA, Mitochondrial; D044402:Receptor, Serotonin, 5-HT2A; D018343:Receptors, Adrenergic, beta-2; D011985:Receptors, Serotonin; D010744:Phosphoric Monoester Hydrolases; C053087:inositol-1,4-bisphosphate 1-phosphatase