AIMS: Genetic association studies have suggested that the single nucleotide polymorphism (SNP) at position 118 of the human mu-opioid receptor (MOR) gene could be a potential risk factor for drug treatment variability in patients. Therefore, we wanted to develop a fast and reliable detection method for this SNP which is applicable in a clinical setting. METHODS: To detect the polymorphism at position A118-->G in the human MOR gene we used the fluorescence resonance energy transfer (FRET)-PCR technique with subsequent melting curve analysis. RESULTS: The polymorphism at position A118-->G in the human MOR gene could be clearly discriminated with melting peak temperatures of 69.8 degrees C and 63.8 degrees C, corresponding to the wild type and mutated MOR allele, respectively. The results from FRET-PCR were validated by sequencing and restriction-fragment length polymorphism (RFLP). Screening of blood samples from 100 subjects showed an allelic distribution for the human MOR alleles of 79% (homozygous wild type), 20% (heterozygous) and 0.9% (homozygous mutated). CONCLUSIONS: The FRET-PCR protocol for detection of the human MOR gene polymorphism at position 118 offers a rapid and reliable method which could be used for population screening of this and other genes.
AIMS: Genetic association studies have suggested that the single nucleotide polymorphism (SNP) at position 118 of the humanmu-opioid receptor (MOR) gene could be a potential risk factor for drug treatment variability in patients. Therefore, we wanted to develop a fast and reliable detection method for this SNP which is applicable in a clinical setting. METHODS: To detect the polymorphism at position A118-->G in the humanMOR gene we used the fluorescence resonance energy transfer (FRET)-PCR technique with subsequent melting curve analysis. RESULTS: The polymorphism at position A118-->G in the humanMOR gene could be clearly discriminated with melting peak temperatures of 69.8 degrees C and 63.8 degrees C, corresponding to the wild type and mutated MOR allele, respectively. The results from FRET-PCR were validated by sequencing and restriction-fragment length polymorphism (RFLP). Screening of blood samples from 100 subjects showed an allelic distribution for the humanMOR alleles of 79% (homozygous wild type), 20% (heterozygous) and 0.9% (homozygous mutated). CONCLUSIONS: The FRET-PCR protocol for detection of the humanMOR gene polymorphism at position 118 offers a rapid and reliable method which could be used for population screening of this and other genes.
Authors: M R Hoehe; K Köpke; B Wendel; K Rohde; C Flachmeier; K K Kidd; W H Berrettini; G M Church Journal: Hum Mol Genet Date: 2000-11-22 Impact factor: 6.150
Authors: M Smolka; T Sander; L G Schmidt; J Samochowiec; H Rommelspacher; N Gscheidel; B Wendel; M R Hoehe Journal: Psychoneuroendocrinology Date: 1999-08 Impact factor: 4.905
Authors: T Funato; Y Nishiyama; N Ioritani; R Matsuki; K Yoshida; M Kaku; T Sasaki; H Ideguchi; J Ono Journal: J Clin Lab Anal Date: 2000 Impact factor: 2.352
Authors: C Bond; K S LaForge; M Tian; D Melia; S Zhang; L Borg; J Gong; J Schluger; J A Strong; S M Leal; J A Tischfield; M J Kreek; L Yu Journal: Proc Natl Acad Sci U S A Date: 1998-08-04 Impact factor: 11.205