Rapid genotyping of the OATP1B1 polymorphisms A388G and T521C with real-time PCR FRET assays.
The polymorphisms (OATP)1B1 A388G and T521C of the solute carrier organic anion-transporter family member 1B1 gene (SLCO1B1), previously known as OATP-C, have potential impacts on drug metabolism. In order to establish a fast and consistent assay for these polymorphisms, rapid speed polymerase chain reaction (PCR) fluorescence resonance energy transfer (FRET) assays on the LightCycler were developed for both OATP1B1 polymorphisms. A locked nucleic acid (LNA) on the polymorphic location within the sensor probe was necessary to discriminate both alleles of the OATP1B1 T521C polymorphism. To confirm the reliability of both real-time PCR FRET assays, these new methods were validated by genotyping 120 samples using a PCR restriction fragment length polymorphism (RFLP) assay and an allele-specific PCR. The results of the real-time PCR FRET assays were completely in line with conventional PCR methods, indicating that the real-time PCR FRET assays are appropriate for clinical settings.
Op den Buijsch
Robert A M
RA
University Hospital Maastricht, Department of Clinical Chemistry, PO Box 58006202, AZ, Maastricht, The Netherlands.
Wijnen
Petal A H M
PA
van Dieijen-Visser
Marja P
MP
de Vries
Johan E
JE
Bekers
Otto
O
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D004247:DNA / Q000235:genetics; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005838:Genotype; D006801:Humans; D027381:Liver-Specific Organic Anion Transporter 1; D008297:Male; D008875:Middle Aged; D027361:Organic Anion Transporters / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D027381:Liver-Specific Organic Anion Transporter 1; D027361:Organic Anion Transporters; C503999:SLCO1B1 protein, human; D004247:DNA