Nucleic acid sequence for detecting diabetes risk site, kit and detection method thereof

Abstract

The invention discloses a nucleic acid sequence for detecting a diabetes risk site, a kit and a detection method thereof. A nucleic acid sequence combination disclosed by the invention is proposed specific to five polymorphic sites (rs228648, rs114202595, rs7754840, rs7756992 and rs7574865) of diabetes, and comprises a wild upstream primer, a mutant upstream primer and a universal downstream primer of each site. The kit for rapidly detecting the diabetes risk site and prepared from the nucleic acid sequence combination has the characteristics of convenience in use, easiness and convenience inoperation, high automation degree, lower pollution in an operation process, good detection effect, high sensitivity, high specificity, high accuracy and high precision. The detection method disclosedby the invention is convenient and rapid to use through adoption of a complete closed-tube operation; a detection result is obtained by directly exploring a fluorescent signal value in a PCR (Polymerase Chain Reaction) process, so that PCR posttreatment or electrophoresis detection is not required, the technical problems of great probability of pollution and false positive value generation in theconventional PCR technology are solved, and non-specific amplification can be effectively avoided; the detection method is suitable for large-scale sample detection.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to a nucleic acid sequence for detecting diabetes risk sites, a kit and a detection method thereof. Background technique [0002] Diabetes mellitus is a polygenic disease in which both genetic and environmental factors participate in the interaction. Both type 1 and type 2 diabetes have obvious genetic heterogeneity. Type 1 diabetes mellitus (T1DM) is an immunomodulatory disease, which is caused by the production of inflammatory cytokines by immune dysregulation, which induces inflammation in pancreatic islets. Type 2 diabetes mellitus (T2DM) is a complex polygenic disease that results in hyperglycemia due to insulin resistance and β-cell secretion defects. More than 90% of diabetic patients have type 2 diabetes. [0003] UST2 (rs228648) affects intracellular Ca after binding to the G protein-coupled receptor GPR14 2+ Therefore, the serum UTS2 of T2DM patients is signifi...

AI Technical Summary

Problems solved by technology

The Sanger sequencing method is the gold standard for mutation analysis, which can find known and unknown mutation sites, but the instrument is expensive, the sensitivity is low, the operation is complicated, the cycle is long, and it is easy to contaminate; the ordinary PCR-RFLP method is simple in technology, cheap in price, and suitable for a small amount of samples laboratory testing, but it can only detect mutations with enzyme cleavage sites, and cannot be detected without enzyme cleavage sites, and there is a risk of false positives caused by PCR product contamination; gene chip technology has the characteristics of high throughput, miniaturization, automation, etc. , suitable for genome-wide mutation scanning, not suitable for mutation site detection of a single gene, low precision, and expensive, these shortcomings limit the development and application of these three methods in actual clinical work, and cannot meet the requirements of rapid clinical evaluation

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