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Method for detecting folic acid metabolic capability based on MassArray nucleic acid mass spectrum and application of method

A technology of folic acid metabolism and ability, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc. It can solve the problems of inability to meet multiple genes, high technical requirements, long detection cycle, etc., and achieve improvement The effect of detection efficiency, shortened detection cycle, and reduced detection cost

Active Publication Date: 2020-12-18
JIANGSU SIMCERE MEDICAL DEVICE CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are many detection methods with their own characteristics. PCR-based detection methods, such as ARMS-PCR, fluorescent quantitative PCR, digital PCR, etc., are easy to operate, fast, and simple in data analysis. However, due to the limitation of throughput, they cannot meet clinical requirements. The need for multiple genes and multiple locus detection
However, high-throughput genetic detection technology, such as high-throughput sequencing technology, solves the limitation of throughput, but its cost is high, the detection cycle is long, and the technical requirements for experimental operations and data analysis personnel are high, so it has not been popularized. applied clinically

Method used

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  • Method for detecting folic acid metabolic capability based on MassArray nucleic acid mass spectrum and application of method
  • Method for detecting folic acid metabolic capability based on MassArray nucleic acid mass spectrum and application of method
  • Method for detecting folic acid metabolic capability based on MassArray nucleic acid mass spectrum and application of method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] The design and optimization of embodiment 1 primer, and the establishment of reaction system

[0075] Through the MassARRAY primer design software (Assay Design Suite), adjust the relevant parameters, complete the PCR and UEP primer pre-design of the three sites, export the designed primers and each parameter file, and synthesize the primers. For fragments with different molecular weights, the detection efficiency of nucleic acid mass spectrometers is different. In order to achieve a more consistent level of detection efficiency at each site, the extension primer UEP in the system needs to be adjusted to an appropriate concentration. The specific method is as follows:

[0076] Dilute 1 μL UEP mix to 25 μL with ultrapure water, perform nucleic acid mass spectrometry detection according to the general procedure, and compare the peak heights of each UEP primer. According to the sample volume and peak height of each UEP primer, the corresponding UEP primers were added acco...

Embodiment 2

[0120] Verification of embodiment 2 reaction system

[0121] In the system optimization process of the present invention, 20 samples of each detection site have been verified by Sanger sequencing, and the comparison results are consistent, confirming that the detection results of the present invention are accurate. After confirming the optimal reaction system, the present invention carries out a series of verification experiments, including accuracy, precision, personnel comparison and comparison experiments between different batches of primers. The specific verification scheme is as follows:

[0122] (1) Accuracy experiment verification plan: 20 samples were used to complete the accuracy experiment, among which serial numbers 1-10 were oral swab samples, and 11-20 were peripheral blood samples (see the table below), and compared Sanger sequencing with The results of MassARRAY, if the consistency is greater than 95%, the verification is passed.

[0123] (2) Precision experim...

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Abstract

The invention provides a primer group and a method for detecting folic acid metabolic capability based on a MassArray nucleic acid mass spectrum platform and application of the method. The primer group for detecting SNP sites related to the folic acid metabolic capability and a detection system of the primer group are designed based on the MassArray nucleic acid mass spectrum platform, so that point mutation related to the folic acid metabolic capability can be accurately typed, and the primer group has the advantages of high accuracy, high sensitivity, good repeatability, low cost, short detection period, intuitive result and the like.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a method for detecting folic acid metabolism capacity based on a MassArray nucleic acid mass spectrometry platform and an application thereof. Background technique [0002] Folic acid is a water-soluble B vitamin composed of pteridine, p-aminobenzoic acid and glutamic acid residues, also known as vitamin BC or vitamin M, which is a substance necessary for the growth and reproduction of body cells and helps protein metabolism , and together with vitamin B12 promote the production and maturation of red blood cells, it is an indispensable substance for the manufacture of red blood cells. Studies at home and abroad have found that the absorption and metabolism of folic acid are related to gene polymorphisms, and the variation of genes will lead to differences in the activity of synthetic enzymes, which will cause differences in the metabolism and absorption of folic acid in the body. T...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/156C12Q2531/113C12Q2533/101C12Q2565/627
Inventor 曹尚志陈永玲任用李诗濛卜范峰丁然吴炳耀
Owner JIANGSU SIMCERE MEDICAL DEVICE CO LTD
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