Primer and method for detecting type of APOE gene

A genotype and sequencing primer technology, applied in the fields of life science and biology, can solve the problems of statin drug efficacy and adverse reactions, etc., and achieve the effect of easy promotion, simple judgment and good effect

Inactive Publication Date: 2018-09-28
南昌艾迪康医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have shown that there are significant individual differences in the pharmacokinetics of statins, and this difference may affect the efficacy of statins and the occurrence of adverse reactions

Method used

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  • Primer and method for detecting type of APOE gene
  • Primer and method for detecting type of APOE gene
  • Primer and method for detecting type of APOE gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 detects the primer of APOE genotype and is combined

[0037] A kind of primer for detecting APOE genotype, the design of this primer is the amplification primer designed for the 4th exon of APOE:

[0038] ApoE-Exon4-F: AACAACTGACCCCGGTGGCG

[0039] ApoE-Exon4-R: ACCTGCTCCTTCACCTCGTC

[0040] The sequencing primers are:

[0041] Sequencing primer 1: AACAACTGACCCCGGTGGCG

[0042] Sequencing primer 2: ACCTGCTCCTTCACCTCGTC

[0043] A test kit for detecting APOE genotype, comprising

[0044] (i) Whole blood genomic DNA extraction reagent;

[0045] (ii) detection system PCR reaction solution;

[0046] (iii) Sequencing system reagents;

[0047] Among them, the extraction of peripheral blood genomic DNA can use the kit of Tiangen Biotechnology Co., Ltd., and the extraction of blood / cell / tissue genomic DNA can refer to the operation process of the kit (Tiangen Biotechnology).

Embodiment 2

[0048] Embodiment 2 detects the method for APOE genotype

[0049] The method for detecting APOE genotype comprises the following steps:

[0050] (1) Genomic DNA extraction from whole blood

[0051] 1) Add 20μl QIAGEN Protease (or proteinase K) to the bottom of a 1.5ml centrifuge tube. .

[0052] 2) Add 200 μl of plasma to the centrifuge tube.

[0053] 3) Add 200 μl Buffer AL and shake for 15 seconds. (Note: Do not add QIAGEN Protease or proteinase K directly to Buffer AL. If the sample size is large, increase QIAGEN Protease and Buffer AL proportionally.)

[0054] 4) Water bath at 56°C for 10 minutes, and then briefly centrifuge to remove the liquid on the inner edge of the centrifuge tube cover.

[0055] 5) Add 200 μl of ethanol (96%-100%), shake for 15 s, and briefly centrifuge to remove the liquid along the inner edge of the centrifuge tube cover.

[0056] 6) Carefully add the mixture obtained above (including the precipitate) to the QIAamp Mini spin column (without wet...

Embodiment 3

[0085] Embodiment 3: APOE genotype detection of clinical samples

[0086] One clinical tissue sample to be tested was taken, and the APOE gene genotype analysis was performed according to the method in Examples 1 and 2, and the results are shown in Tables 4 and 5. Such as Figure 4 As shown, the 388th base position is T. Such as Figure 5 As shown, the 526th base position is C. comprehensive Figure 4 with Figure 5 The results showed that the APOE genotype was E3.

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Abstract

The invention discloses a primer and method for detecting the type of an ApoE gene related with cardiovascular diseases such as atherosclerosis and coronary heart disease, and Alzheimer disease. The primer is used for amplifying the fourth exon sequence of the gene, and a Sanger sequencing technique is adopted. The base group condition of the encoded 112th and 158th amino acid sequences of the ApoE gene can be quickly detected so as to determine the genotype. The detected results obtained through the primer and the method is accurate, and the important reference significance on early diagnosisof atherosclerosis and Alzheimer disease and individualized treatment is achieved.

Description

technical field [0001] The invention belongs to the field of life science and biotechnology, and particularly relates to a primer and a method for detecting APOE genotypes related to atherosclerosis and senile dementia. Background technique [0002] Apolipoprotein E (Apolipoprotein E, ApoE) is a protein of 299 amino acids with a molecular weight of 34 kDa. Is a major cholesterol carrier, has the function of supporting lipid transport and repairing brain damage. The ApoE gene is located on the long arm of chromosome 19, with a total length of about 3.7kb and 4 exons. The 4th exon exhibits gene polymorphism, which makes the ApoE gene have E2, E3, and E43 alleles. The combination produced six genotypes: three homozygous (E2 / 2, E3 / 3, E4 / 4) and three heterozygous (E3 / 2, E4 / 2, E4 / 3). The ApoE gene has isoform specificity, that is, the protein functions encoded by different genes are different, resulting in different proteins' different ability to metabolize lipids, and different...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/156
Inventor 董绍斌吴鹏飞王淑一
Owner 南昌艾迪康医学检验实验室有限公司
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