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Primer set, kit and method for detecting monogenic diabetes related genes

A diabetes, single-gene technology, applied in the field of primer sets, can solve the problems of limited clinical application and high testing costs.

Active Publication Date: 2021-06-01
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, even under the circumstances that the current sequencing cost is decreasing year by year, the cost of the test is still high, and the cost ranges from 1000-5000 yuan
This also seriously limits the clinical application

Method used

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  • Primer set, kit and method for detecting monogenic diabetes related genes
  • Primer set, kit and method for detecting monogenic diabetes related genes
  • Primer set, kit and method for detecting monogenic diabetes related genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] The present embodiment is used to illustrate the method for detecting the pathogenic gene of single-gene diabetes mellitus using the primer combination of the present invention, and the specific steps are as follows:

[0091] Step A: Sample

[0092] For blood samples, saliva or other tissue samples obtained from hospitals or other research institutions, use a DNA extraction kit to extract whole genome DNA, and use a nucleic acid quantifier to measure the concentration of the extracted DNA. The DNA concentration is 50ng / ul, and the total amount is not. Below 200ng.

[0093] For the above samples, configure the PCR reaction system, for LMNA, PPP1R15B, KLF11, DCAF17, NEUROD1, PPARG, APPL1, WFS1, TRMT10A, CISD2, PIK3R1, ZFP57, RFX6, ZBTB2, GCK, PAX4, BLK, GATA4, CEL, PCBD1, SLC29A3, INS, ABCC8, KCNJ11, PAX6, HNF1A, PDX1, DNAJC3, PLIN1, HNF1B, GATA6, INSR, DYRK1B, AKT2, POLD1, HNF4A and MTTL1. For amplification, MJ1909CL37 Panel Mix A and MJ1909CL37 Panel Mix B were used ...

Embodiment 2

[0167] In order to verify the stability of the system and the accuracy of the typing results, we extracted DNA from blood samples from 6 random diabetic patients using the commonly used and recognized Agilent exome sequencing kits and the genes described in this patent, respectively. The detection kits were used for library construction and sequencing, and then for mutation site detection and analysis. The final result shows that the genotype of the target region obtained by this kit is exactly the same as the genotype of this region in the Agilent exome sequencing kit. It shows that the accuracy of this method for this region is comparable to that of Agilent and other whole-exome sequencing. Only the detected mutation sites are shown below.

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Embodiment 3

[0182] In order to verify the repeatability of the system, we carried out two repetitions of blood samples from 10 different diabetic patients. The final analysis results showed that the mutation sites found in the two experiments were exactly the same. Only the detected mutation sites are shown below.

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[0214] It is illustrated that the detection method provided by the present invention has good detection balance, high sensitivity, good repeatability and accurate typing results, and can be used as an auxiliary diagnostic tool for mon...

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PUM

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Abstract

The application discloses a primer set, a kit and a method for detecting the pathogenic gene of monogenic diabetes. Through multiple amplification combined with next-generation sequencing and the experimental and analysis methods created for this system, in one PCR reaction, the exons of 37 genes and the 50bp regions on the left and right sides can be detected, mutation sites can be found, and Annotate the results of mutation sites, and carry out disease typing at the gene level, so as to achieve the purpose of auxiliary diagnosis. The method is simple, easy, fast, efficient and low in cost, and provides a feasible and simple screening method for clinical screening of monogenic diabetes.

Description

technical field [0001] The present invention relates to a primer set for amplifying genes related to monogenic diabetes. [0002] The present invention relates to a kit for detecting genes related to monogenic diabetes. [0003] The present invention relates to a method for detecting the related genes of monogenic diabetes. Background technique [0004] Monogenic diabetes mainly includes neonatal diabetes, adult-onset juvenile diabetes (MODY), mitochondrial diabetes and syndromic rare diabetes. The incidence of these diseases is very low, the incidence of neonatal diabetes is about 1 in 200,000, and MODY accounts for about 1%-3% of the total diabetic population. Due to the early onset, severe symptoms, and large population base, even if the incidence rate is low, the total number of patients is very high: about 8 million people in my country suffer from monogenic diabetes, which is a number that cannot be ignored and has become an urgent problem in clinical diagnosis and t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/112C12Q2600/156C12Q2600/16C12Q2531/113C12Q2535/122C12Q2537/143
Inventor 周凯欣徐涛郑丽王友赵晶
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES