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Primer set, application, product and method for detecting related SNP sites of drug metabolism ability of rheumatic immune diseases
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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. Effect
Active Publication Date: 2020-07-28
JIANGSU SIMCERE MEDICAL DEVICE CO LTD +2
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[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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Embodiment 1
[0100] Example 1: Design and optimization of primers, and establishment of reaction system
[0101] Through the primer design software (Assay Design Suite) on the MassARRAY website, adjust the relevant parameters to complete the PCR and UEP primer design of 44 sites (including a repeat detection site rs2231142 and gender identification site 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. Then proceed to primer testing and optimization. Specific steps are as follows:
[0102] (1) Dilute the genomic DNA sample to 10ng / μL, and prepare the PCR reaction MIX as shown in the following table, where PCR PrimerMix is two sets of different amplification primer mixtures (the following is a single sample volume):
[0103] Table 3 PCR reaction system
[0104]
[0105] Seal the film,...
Embodiment 2
[0151] Example 2 Verification of the reaction system
[0152] In the process of system optimization, each detection site has 2 to 3 samples verified by Sanger sequencing, and the comparison results are consistent, confirming that the detection results are accurate. After confirming the optimal reaction system, perform the following experiments:
[0153] (1) Accuracy experimental verification scheme: 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) Accuracy experimental verification plan: pick 2 exceptions of weekly blood samples and 2 oral swab samples, each sample is repeated 3 times for a batch of testing, a total of 3 batches are tested, precision and batch If the consistency of the internal precision is greater than 95%, the verification is passed;
[0155] (3) Experimental operator comparison and reagent comparison experim...
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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 relevant SNP sites of drug metabolism in rheumatic immune diseases. Background technique [0002] Rheumatic immune disease is a general term for a large group of diseases. The most common clinical diseases mainly include rheumatoid arthritis, systemic lupus erythematosus, ankylosing spondylitis, gout, etc. Rheumatic immune disease is a persistent and recurrent disease. Most patients need to live with the disease for a long time. There is currently no cure for most of the diseases. The treatment is aimed at controlling the disease through drugs, maintaining clinical remission, and improving the quality of life. However, it is a pity that even with regular treatment under the guideline standard drug treatment plan, the disease control rate and remission rate are still very low. [0003] Although in recent decades, with peopl...
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