A simultaneous LightCycler detection assay for five genetic polymorphisms influencing drug sensitivity

Masahiro Hiratsuka, Kaori Narahara, Yukinaga Kishikawa, Samar Ismail Hamdy, Naomi Endo, Yasuyuki Agatsuma, Masaki Matsuura, Tomoko Inoue, Yoshihisa Tomioka, Michinao Mizugaki

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Objectives: 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. Design and 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.

Original languageEnglish
Pages (from-to)35-40
Number of pages6
JournalClinical Biochemistry
Volume35
Issue number1
DOIs
Publication statusPublished - 2002 Apr 17

Keywords

  • Drug sensitivity
  • Pharmacogenetics
  • SNP

ASJC Scopus subject areas

  • Clinical Biochemistry

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