Primer group, application, product and method for detecting SNP (single nucleotide polymorphism) loci related to drug metabolism capacity of rheumatic immunological diseases

A technology of metabolic capacity and rheumatic immunity, which is applied in the fields of biochemical equipment and methods, recombinant DNA technology, and microbial determination/inspection, etc., can solve problems such as lack of throughput, and achieve lower detection costs, higher accuracy, and shorter detection cycles. Effect

Active Publication Date: 2019-12-10
JIANGSU SIMCERE MEDICAL DEVICE CO LTD +2
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  • Abstract
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Problems solved by technology

[0010] The third object of the present invention is to provide a product for the detection of individualized medicines for rheumatic immune diseases, so as to alleviate the lack of high-throughput, easy-to-operate and low-cost detection products for individualized medicines for rheumatic immune diseases in the prior art. Product technical issues

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  • Primer group, application, product and method for detecting SNP (single nucleotide polymorphism) loci related to drug metabolism capacity of rheumatic immunological diseases
  • Primer group, application, product and method for detecting SNP (single nucleotide polymorphism) loci related to drug metabolism capacity of rheumatic immunological diseases
  • Primer group, application, product and method for detecting SNP (single nucleotide polymorphism) loci related to drug metabolism capacity of rheumatic immunological diseases

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Effect test

Embodiment 1

[0100] Example 1: Design and optimization of primers, and establishment of reaction system

[0101] Through the primer design software (Assay Design Suite) of the MassARRAY website, adjust the relevant parameters, complete the PCR and UEP primer design of 44 loci (including a repeat detection locus rs2231142 and sex identification locus AMEL), and export the designed primers And each parameter file, and synthesize primers. Prepare amplification primer MIX and extension primer MIX according to the primer configuration table, and fine-tune the extension primer MIX until it meets the requirements. Primer testing and optimization were then performed. Specific steps are as follows:

[0102] (1) Dilute the genomic DNA sample to 10ng / μL, and prepare the PCR reaction MIX according to the following table, where the PCR PrimerMix is ​​a mixture of two sets of amplification primers with different sites (the following is the amount of a single sample):

[0103] Table 3 PCR reaction sys...

Embodiment 2

[0151] Verification of embodiment 2 reaction system

[0152] In the process of system optimization, 2 to 3 samples of each detection site were verified by Sanger sequencing, and the comparison results were consistent, confirming that the detection results were accurate. After confirming the optimal reaction system, the following experiments were carried out:

[0153] (1) Accuracy experimental verification plan: select one sample from each of the 43 sites for Sanger sequencing, and compare the results of Sanger sequencing and MassARRAY. If the consistency is greater than 95%, the verification is passed;

[0154] (2) Precision experiment verification plan: pick 2 peripheral blood samples and 2 oral swab samples, repeat 3 times for each sample to conduct a batch of detection, a total of 3 batches were detected, the precision between batches and batches If the consistency of internal precision is greater than 95%, the verification is passed;

[0155] (3) Experimental operator co...

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Abstract

The invention provides a primer group, an application, a product and a method for detecting SNP (single nucleotide polymorphism) loci related to drug metabolism capacity of rheumatic immunological diseases, and relates to the technical field of biology. The primer group for detecting the SNP loci related to the drug metabolism capacity of the rheumatic immunological diseases can be used for detecting metabolism capacity of an immunosuppressor, metabolism capacity of an immunomodulator, metabolism capacity of an anti-inflammatory agent, metabolism capacity of a TNF inhibitor, metabolism capacity of a B cell regulator, metabolism capacity of an IL-6 inhibitor, metabolism capacity of a uric acid lowering agent and metabolism capacity of a uricosuric agent. The primer group is designed for main mutation sites related to common drug metabolism of the rheumatic immunological diseases, can realize specific detection for locus mutation related to common drug metabolism of the rheumatic immunological diseases, and can greatly shorten detection cycle and reduce detection cost.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a primer set, application, product and method for detecting related SNP sites of drug metabolism ability of rheumatic immune diseases. Background technique [0002] Rheumatic immune disease is a general term for a large class of diseases, the most common clinically including rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, gout and so on. Rheumatic immune diseases are protracted and recurring diseases. Most patients need to live with the disease for a long time, and there is no radical cure for most diseases. The purpose of treatment is to control the disease through drugs, maintain clinical remission, and improve the quality of life. But unfortunately, even with regular treatment under the guideline standard drug regimen, the disease control rate and remission rate are still very low. [0003] Although in recent decades, with the recognition of di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 赵妍李翔陈珊珊李小方李杜衡肖念清任用
Owner JIANGSU SIMCERE MEDICAL DEVICE CO LTD
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