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Individual nutrients deficiency risk assessment method based on SNP (Single Nucleotide Polymorphism) site

A risk assessment and risk assessment model technology, applied in the field of human nutrition assessment, can solve problems such as unfavorable automation and long running time, and achieve the effect of ensuring accuracy, strong ductility, and flexible customization

Inactive Publication Date: 2018-05-18
NAT INST FOR NUTRITION & HEALTH CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of operation process, this method involves three major steps of conventional PCR amplification, substrate removal and single base extension. There are many kinds of chemical reagents and long running time. It requires professionals to design primers, operate equipment and analyze results, so it is not conducive to the realization of automation

Method used

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  • Individual nutrients deficiency risk assessment method based on SNP (Single Nucleotide Polymorphism) site
  • Individual nutrients deficiency risk assessment method based on SNP (Single Nucleotide Polymorphism) site
  • Individual nutrients deficiency risk assessment method based on SNP (Single Nucleotide Polymorphism) site

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Extraction and quality control of embodiment 1 human DNA

[0050] Use a DNA extraction kit or salting-out method to extract human DNA from blood or other tissues according to the instructions, and use a DNA detector or (and) agarose gel electrophoresis to test the quality of the DNA sample.

Embodiment 2

[0051] Design and synthesis of embodiment 2 SNP site primers

[0052] The first primer mixture contains two specific upstream primers and one universal downstream primer, and the fluorescent substance mixture contains two primers with fluorescent groups. The SNP site in the primer mixture is located at the last base at the 3' end of the upstream primer, and the tag sequences at the 5' end of the two specific upstream primers are different. During PCR amplification, the two specific primers competitively bind. The two primers with fluorescent groups in the fluorescent substance mixture are complementary to the 5' tag sequences of the two specific upstream primers in the primer mixture. Before the PCR reaction, the fluorescent groups (FAM or VIC) do not fluoresce and cannot be detected. Signal, during the PCR process, a sequence that is completely complementary to the upstream primer is synthesized, the fluorescent primer can be combined with the DNA sequence that is completely ...

Embodiment 3

[0080] Design and synthesis of embodiment 3SNP site primers

[0081] 1. Pre-spot primers or DNA on each well of the chip or well plate using a manual or automatic spotting instrument.

[0082] 2. Inject the master mix into the injection hole of the chip or each well of the orifice plate

[0083] The master mix includes 4 μl of DNA or primer mixture with a concentration of 2.5-50 ng / μl, 10 μl of fluorescent substance mixture, and 6 μl of ultrapure water. All components need to be vortexed and mixed evenly before use, and then the reaction master mix is ​​manually added from the chip injection hole or automatic sampler to the chip or well plate. For 96-well plate, 384-well plate and 1536-well plate, the volume of each individual reaction well is 10μl, 5μl and 1μl.

[0084] 3. Seal the membrane and centrifuge

[0085] Blocks were sealed and centrifuged. Each type of orifice plate is sealed and centrifuged manually or by heat sealing machine and (or) laser sealing machine.

...

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Abstract

The invention discloses an individual nutrients deficiency risk assessment method based on SNP (Single Nucleotide Polymorphism) site, which is named as INRAS (Individual nutrients deficiency risk assessment based on SNP). The method is aimed at N SNP site of an individual or N individuals of an SNP site as an experimental unit, the studied object is SNP site related to individual nutrient metabolism capacity, and primers are designed and synthesized according to locus information; DNA is then extracted out from the blood of the individual to carry out PCR (polymerase chain reaction) amplification, an assay result is then obtained by a fluorescence scanner, and finally, a risk assessment model is adopted to analyze the result. The risk assessment model disclosed by the invention, which is created on an evidence-based inclusion basis of SNP site, can accurately and systematically assess various nutrients deficiency risks of the human body on the basis of the SNP site; and a dynamic riskassessment model which is created on the basis of critical values of biochemical indexes of population nutrients can carry out accurate and systematic dynamic assessment on various nutrients deficiency risks of populations on the basis of the SNP site.

Description

technical field [0001] The invention belongs to the technical field of human nutrient assessment, and specifically relates to a method for assessing the risk of human nutrient deficiency based on SNP sites, also known as the INRAS method. Background technique [0002] Single nucleotide polymorphism (single nucleotide polymorphism, SNP) refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genome level. It is an important step towards the application of the Human Genome Project and can be used for nutrient metabolism. Discovery of high-risk groups. After restriction enzyme fragment fragment length polymorphism and short tandem repeat sequence, single nucleotide polymorphism site (SNP) has become the third place because of its high density of genetic markers, high stability, and easy automation of typing detection. Generation polymorphic markers have shown strong application prospects in the fields of human genomics research such as gen...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/156
Inventor 张春红霍军生孙静黄建
Owner NAT INST FOR NUTRITION & HEALTH CHINESE CENT FOR DISEASE CONTROL & PREVENTION
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