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Primers, probes and kit for detecting polymorphisms of ApoE gene and SLCO1B1 gene

A gene polymorphism and kit technology, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of instrument temperature control, high false positives, cumbersome steps, etc., and achieve high sensitivity , strong specificity and simple operation

Active Publication Date: 2017-03-29
武汉海吉力生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the DNA sequencing method is the gold standard, the steps are cumbersome and time-consuming
High-resolution melting curve method is fast, simple, economical and practical, but it is controlled by the temperature of the instrument, and the false positive is high

Method used

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  • Primers, probes and kit for detecting polymorphisms of ApoE gene and SLCO1B1 gene
  • Primers, probes and kit for detecting polymorphisms of ApoE gene and SLCO1B1 gene
  • Primers, probes and kit for detecting polymorphisms of ApoE gene and SLCO1B1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1 kit composition

[0074] There are two types of detection kits listed in this example, see Table 1 and Table 2 respectively.

[0075] Table 1 Kit composition one

[0076]

[0077]

[0078] Table 2 Kit composition two

[0079]

Embodiment 2

[0080] Embodiment 2: Extraction of clinical sample DNA

[0081] 1. EDTA anticoagulant

[0082] In this example, genomic DNA was extracted from EDTA anticoagulated blood and quantified as a template for PCR detection. The blood genomic DNA extraction kit of Tiangen was used, and the specific details are as follows.

[0083] 1. Processing blood material:

[0084] a. When the volume of the blood sample is less than 200 μL, add buffer GS to make up the volume to 200 μL, and then proceed to the next step of the experiment (if the volume of the blood sample is 200 μL, the next step of the experiment can be carried out directly without adding GS).

[0085] b. When the volume of the blood sample exceeds 200 μL, it needs to be treated with cell lysate CL. The specific steps are as follows: add 1 to 2.5 times the volume of cell lysate CL to the sample, mix it upside down, and centrifuge at 10,000 rpm (~11,500×g) After 1 min, remove the supernatant and leave the cell nucleus pellet (i...

Embodiment 3

[0113] Embodiment 3: Real-time fluorescence PCR method amplifies clinical sample DNA

[0114] In this example, the primers and probes provided in Example 1 are used to amplify the DNA sample extracted in Example 2.

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PUM

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Abstract

The present invention discloses primers, probes and a kit for detecting the polymorphisms of gene ApoE and gene SLCO1B1, wherein the primers and the probes comprise primers and probes for detecting the polymorphic loci of T388C and C526T of the gene ApoE, and primers and probes for detecting the polymorphic loci of A388G and T521C of the gene SLCO1B1. According to the present invention, the kit is suitable for the selection of a variety of clinical samples, and has significant advantages of high specificity, high sensitivity, short experimental period, simple operation, safety, no toxicity, low cost, and the like.

Description

technical field [0001] The invention relates to the technical field of gene polymorphism detection, in particular to primers, probes and kits for detecting ApoE and SLCO1B1 gene polymorphisms. Background technique [0002] Statins are hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, the most classic and effective lipid-lowering drugs, and are widely used in the treatment of hyperlipidemia. At the same time, early application in patients with acute coronary syndrome can inhibit the inflammatory response of vascular endothelium, stabilize atherosclerotic plaque, improve vascular endothelial function, delay the degree of atherosclerosis (AS), anti-inflammatory, nerve protection and antithrombotic And so on. [0003] The human apolipoprotein E (ApoE) gene is located on chromosome 19 and has 4 exons and 3 introns. The two most common mutations of the ApoE gene are T388C and C526T, which can form 3 haploids, respectively E2 (388T-526T), E3 (388T-526C), E4 (388C-5...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6827C12Q1/6883C12Q2600/106C12Q2563/107C12Q2545/114C12Q2531/113
Inventor 吴志伟段卫涛史学辉裴冰雪胡利红赵平锋卢彦羽肖红涛张全
Owner 武汉海吉力生物科技有限公司
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