Rapid and cost effective genotyping method for polymorphisms in PPARG, PPARGC1 and TCF7L2 genes.
Polymorphisms (rs1801282, rs8192678, rs7903146) of peroxisome proliferator-activated receptor gamma (PPARG), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A) and transcription factor 7-like 2 (TCF7L2) have recently been associated with different diseases, mainly type 2 diabetes. An assay using unlabeled probes and the LightCycler or Rotor-Gene instruments was developed for genotyping of these three polymorphisms. Asymmetric polymerase chain reaction was used, followed by melting analysis of the unlabeled probe/ssDNA amplicon duplex. Samples with the target genotypes were accurately detected and easily distinguishable. Thus, genotyping using unlabeled probes is a rapid, accurate and cost effective closed-tube method. These assays demonstrated 100% specificity and sensitivity for the identification of selected polymorphisms in PPARG, PPARGC1A and TCF7L2 genes.
Habalová
Viera
V
Department of Medical Biology, Faculty of Medicine, Safárik University, Tr. SNP 1, 040 66 Kosice, Slovakia. viera.habalova@upjs.sk
Klimcáková
Lucia
L
Zidzik
Jozef
J
Tkác
Ivan
I
D003362:Cost-Benefit Analysis; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D047495:PPAR gamma / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000191:economics* / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D051820:TCF Transcription Factors / Q000235:genetics*; D058118:Transcription Factor 7-Like 2 Protein; D014157:Transcription Factors / Q000235:genetics*
D047495:PPAR gamma; D051820:TCF Transcription Factors; C544894:TCF7L2 protein, human; D058118:Transcription Factor 7-Like 2 Protein; D014157:Transcription Factors; C111264:peroxisome-proliferator-activated receptor-gamma coactivator-1