Primers and method for detecting related gene SNP (single nucleotide polymorphism) sites of metformin-individualized medications

Abstract

The present invention relates to primers and a method for detecting related gene SNP (single nucleotide polymorphism) sites of metformin-individualized medications. The primers can detect a rs2289669SNP site of a SLC47A1 gene. The primer sequences comprise a wild-type upstream primer, a mutant-type upstream primer and a shared downstream primer. Improvement is made on the basis of ARMS-PCR, a specific primer and the shared downstream primer are designed for SNP sequences, mismatched bases are introduced at a second position of a 3' end of the specific primerto widen a gap between amplification efficiency of complementation and non-complementation of the bases of the 3' end of the specific primer and SNP site bases on a DNA template, thereby greatly improving specificity and accuracy of detection. A real-time PCR technology is introduced, identification can be made according to a [delta]Ct value, that is, the [delta]Ct value is equal to mutant Ct-wild Ct. In general, when [delta]Ct islarger than 7, the SNP site is a wild type, when [delta]Ct is less than -7, the SNP site is a mutant type, and when the [delta]Ct is larger than -2 and less than 2, the SNP site is a heterozygous type. Advantages are that the method is fast, simple, economical, accurate and reliable identification of results, and high in specificity.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a primer and a method for detecting SNP sites of genes related to metformin individualized medication. Background technique [0002] Diabetes mellitus is an endocrine and metabolic disease caused by insulin deficiency or reduced biological effects, and its incidence is increasing year by year. According to the World Health Organization, there will be 380 million diabetic patients in the world in 2025. On January 9, 2012, the China Health Education Center announced that "China's chronic disease monitoring and diabetes prevalence rate is 2.6%, and the prevalence rate of the elderly over 60 years old is as high as 19.6%. There are 97 million adult diabetic patients in the country, surpassing India. It has become the country with the largest number of diabetics in the world”, which has caused a heavy burden on the physical and mental health of patients and the economy. ...

AI Technical Summary

Problems solved by technology

ARMS-PCR can detect any point mutations and small deletions on DNA molecules. Its method is reliable and does not depend on any special markers. However, gel electrophoresis is often required after the reaction, which increases the chance of PCR contamination and is time-consuming and laborious.

Images