Comparison of two screening methods for in-house genotyping in clinical pharmacology units.
Two genetic screening methods, the fluorescence resonance energy transfer (FRET) technique on the LightCycler and the real-time pyrophosphate detection technique on the Pyrosequencer have been compared with regard to their usefulness as screening methods for subject recruitment in clinical studies in pharmacology units. Two SNPs of possible clinical relevance were selected, namely the 118A>G SNP of the OPRM1 gene and the 3435C>T SNP of the ABCB1 gene. Genotypes diagnosed using conventional sequencing served as control. The allelic frequency of the mutated 118G allele of the OPRM1 gene was 12.7% and that of the mutated 3435T allele of the ABCB1 gene was 50.7%. All results obtained with the Pyrosequencer were in accord with those obtained using conventional sequencing. With the LightCycler, an incorrect genotype was assigned to 1 of the 130 DNA samples corresponding to an error rate of 0.8%. Although both methods were found suitable for rapid SNP detection, Pyrosequencing was the preferred method since it provides the nucleotide sequence directly thus facilitating interpretation.
Darimont
J
J
pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Johann Wolfgang Goethe-University, Frankfurt/Main, Germany.
Grösch
S
S
Skarke
C
C
Geisslinger
G
G
Lötsch
J
J
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D002986:Clinical Trials as Topic; D004247:DNA / Q000032:analysis; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods* / Q000592:standards; D005820:Genetic Testing / Q000379:methods; D005838:Genotype; D006801:Humans; D018579:Patient Selection; D010600:Pharmacology; D010710:Phosphates / Q000032:analysis; D020641:Polymorphism, Single Nucleotide; D017450:Receptors, Opioid, mu / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D018528:ATP-Binding Cassette Transporters; C485764:OPRM1 protein, human; D010710:Phosphates; D017450:Receptors, Opioid, mu; D004247:DNA