Design of an allele-specific PCR assay to genotype the rs12255372 SNP in a pilot study of association between common TCF7L2 polymorphisms and type 2 diabetes in Venezuelans

Abstract

Objective: The global burden of diabetes mellitus will impact strongly American countries in the coming decades. Type 2 diabetes mellitus (T2DM) is a multifactorial disease and the basis for its genetic susceptibility remains not fully understood. Different population studies have demonstrated that variants of the TCF7L2 gene are strongly associated with an increased risk of T2DM. Moreover, institutions or countries with limited budget to conduct genetic research need cost effective methods for detecting DNA variants. Subjects and methods: We standardized a rapid and simple allele-specific PCR method for genotyping the rs12255372 single nucleotide polymorphism (SNP) in a pilot study exploring the association of three TCF7L2 polymorphisms (rs7903146, rs12255372 and DG10S478) with T2DM in 70 patients and 73 controls from Venezuela. Results: The performance of the designed allele-specific PCR reaction for rs12255372 genotyping was reliable and accurate. Patients carrying the TCF7L2 rs7903146 T allele (CT + TT genotypes) and heterozygous CT genotype had a significantly higher risk for T2DM (OR = 2.9 and 2.3, respectively). Although rs12255372 and DG10S478 risk alleles predominated in T2DM group no statistical significance was found. Conclusions: We developed a novel allele-specific PCR method for easier and rapid detection of rs12255372 polymorphism without the use of expensive instrumentation and reagents. Our study in a relatively small sample of the Venezuelan population replicated the association of the rs7903146 SNP with T2DM. Further studies with larger sample size and more biochemical data should be conducted to explore the genetic basis of T2DM susceptibility in Venezuela. Arch Endocrinol Metab. 2016;60(3):246-51

Pilot studies on the prevalence and association of TCF7L2 polymorphisms in non-diabetic participants and type 2 diabetic patients of South Tamil Nadu

“…In contrast to the association of the rs7903146 SNP with lean body mass, the rs12255372 GT/TT genotype has been strongly associated with metabolic syndrome/ increased waist circumference. [23][24][25] Hence it could be suggested that the present study data may fall in line with these reports, as the rs12255372 positive participants tended to exhibit a relatively greater BMI and abnormal OGTT results.…”

Multiplex signal cascade fluorescent biosensing for single nucleotide polymorphisms enhanced detection

“…Thus, the accurate and effective SNPs of IDH-1 gene detection make glioma therapy more promising. Traditional strategies for identification and typing of SNPs mainly involved microarray analysis (Divne and Allen, 2005), allele-specific PCR (Moran et al, 2016) and sequencing (Zhou et al, 2013), but they had not been promoted or prevailed due to the drawbacks of high false-positive rate, low sensitivity and multi-step washing (Zhu et al, 2014). Therefore, it is urgent to establish prominent sensing for SNPs sensitive, accurate and portable detection (Knez et al, 2014).…”