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SURVEYING VARIANTS RS757110 IN TYPE 2 DIABETES PATIENTS BY ARMS-PCR
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Abstract
Background: Type 2 diabetes is a chronic disease with genetic predisposition. Several studies have shown the association between type 2 diabetes and certain single polymorphisms, including rs757110 of ABCC8 gene. Sanger sequencing, so far, is the gold standard for determining single nucleotide polymorphism, but it is costly and time consuming. Therefore, it is necessary to develop a method that provides accurate information and is simple enough to be widely applied. Objectives: To develop a protocol to determine single nucleotide polymorphism variants rs757110 in patients with type 2 diabetes. Methods: The study was carried out on DNA samples isolated from the blood of 50 patients with type 2 diabetes and 50 controls. The rs757110 variant was identified by Tetra-ARMS PCR combined with Sanger sequencing. Results: Rs757110 was successfully identified by tetra-ARMS PCR. Rs757110 was in Hardy-Weinberg equilibrium with p-value = 0,83. The G allele frequencies were 0,44 and 0,27 in the patient and control samples, respectively. The G/G genotype increased the risk of type 2 diabetes compared with the T/T genotype (OR=9,67, 95%CI=1,92-48,8, p<0,05). The T/G genotype increased the risk of the disease compared with the T/T genotype (OR=3,23, 95%CI=1,11-9,39, p<0,05). Conclusion: We have successfully built a procedure to identify the single nucleotide polymorphism variants rs757110 with high sensitivity and specificity. This variant has been shown to be associated with type 2 diabetes.
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Keywords
type 2 diabetes, rs757110, ABCC8, Tetra-ARMS PCR
References
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