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Complex disease-related SNP site primer composition and use thereof

A primer composition and disease technology, applied in the field of molecular biology, can solve the problem of not providing a variety of complex disease detection methods or systems, and achieve the effects of eliminating non-specific amplification, simple operation and good sensitivity

Inactive Publication Date: 2021-09-17
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above prior art only provides an evaluation method or system for a certain complex disease, and does not provide a detection method or system for multiple complex diseases

Method used

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  • Complex disease-related SNP site primer composition and use thereof
  • Complex disease-related SNP site primer composition and use thereof
  • Complex disease-related SNP site primer composition and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Primer Composition

[0070] The primer composition described in this application is specifically shown in Table 2 below.

[0071] Table 2 Primer Composition

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[0073]

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[0075]

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Embodiment 2

[0083] Example 2 library construction and detection

[0084] 1. Genomic DNA Extraction

[0085] Genomic DNA (gDNA) was extracted from saliva or blood according to the operating steps of the commercial genomic DNA extraction kit, and quantified using Qubit (Life Technologies).

[0086] 2. The first round of multiplex PCR reaction

[0087] 1.1. The reaction system is shown in Table 3 below.

[0088] Table 3 reaction system

[0089] Reagent Volume(μL) wxya 2 o

4-x Enhancer NB(1N) 5 Primer (Table 2) 2.4 gDNA X (30ng) IGT-EM101polymerase mixture 3.6

[0090] Wherein, the final concentration of each primer in the Primer primer mixture is 0.5 pmol.

[0091]1.2. The reaction program is: 95°C for 3min30s; 98°C for 20s, 60°C for 8min, 18cycle; 72°C for 5min.

[0092] 3. The first round of magnetic bead purification

[0093] 3.1. Add 15 μL of AMPure XP magnetic beads equilibrated at room temperature to 15 μL of PCR product, and mix...

Embodiment 3

[0128] The corresponding relationship between complex diseases and genes and detection sites is shown in Table 5 below.

[0129] table 5

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[0137] Experimental Example 1 Accuracy Detection

[0138] The detection method described in this application was used to detect the above 221 sites, and at the same time, Sanger sequencing was used for verification. The results are shown in Table 6 below.

[0139] Table 6 Accuracy test results

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[0146] It can be seen from Table 6 that the accuracy of the primer composition and detection method described in this application is 100%.

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Abstract

The invention provides a complex disease-related SNP site primer composition and use thereof, and belongs to the technical field of molecular biology. A group of SNP site amplification primers are designed and developed through an NGS method. A change of a gene structure related to complex disease in an individual body is detected through a bioinformatics analysis method. The complex disease-related SNP site primer composition predicts an individual disease risk and improves prevention and / or treatment effects of complex disease. The evaluation system and the detection method have low cost, are simple and convenient to operate, and have good sensitivity, high accuracy, good repeatability and great clinical application value.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a complex disease-related SNP site primer composition and application. Background technique [0002] Complex disease is a general term for a class of diseases with complex etiology, insidious onset, long course and protracted disease. In recent years, with the process of rapid urbanization and population aging, the morbidity and mortality of chronic complex diseases have shown a continuous upward trend. Epidemiological surveys have shown that the occurrence of chronic complex diseases is closely related to unhealthy lifestyles and behaviors. The monitoring of chronic and complex diseases can detect the health problems of the body in time, and propose corrective measures for related problems. Early screening and detection of chronic complex diseases is of great significance for reducing the harm of chronic complex diseases. To prevent the occurrence of chronic complex ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/6869C12Q2600/156C12Q2531/113C12Q2537/143C12Q2523/308
Inventor 方向东单广乐杨亚东隋阳辛子娟
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
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