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A rapid screening method for a single nucleotide polymorphism (SNP) in the human MOR gene
Author(s) -
Grösch Sabine,
Niederberger Ellen,
Lötsch Jörn,
Skarke Carsten,
Geisslinger Gerd
Publication year - 2001
Publication title -
british journal of clinical pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.216
H-Index - 146
eISSN - 1365-2125
pISSN - 0306-5251
DOI - 10.1046/j.0306-5251.2001.01504.x
Subject(s) - single nucleotide polymorphism , genetics , biology , allele , restriction fragment length polymorphism , polymorphism (computer science) , snp , gene , microbiology and biotechnology , polymerase chain reaction , allele frequency , population , melting curve analysis , genotype , medicine , environmental health
Aims Genetic association studies have suggested that the single nucleotide polymorphism (SNP) at position 118 of the human µ‐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 ° C and 63.8 ° 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.