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Kit and method for detecting hypertension drug related gene polymorphism

A gene polymorphism and kit technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of high sequencing cost, labor-intensive and material resources-consuming detection time, and long time. Achieve the effect of reducing the cost of reagent consumables and labor costs, reducing experimental steps and inspections, and simple operation

Pending Publication Date: 2019-11-01
广州海思医疗科技有限公司
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AI Technical Summary

Problems solved by technology

[0008] (1) Hypertension is generally accompanied by complications of cardiovascular and cerebrovascular diseases, and most patients need combined medication. Products on the market generally only target the detection of one or more hypertension drug genes, and do not include related complications drug;
[0009] (2) Although the cost of next-generation sequencing technology has dropped significantly, the price of testing is still an important factor restricting its large-scale application in the market
Among them, the complex process of building a library, a long time, and the consumption of manpower and material resources are important reasons for the long detection time and high sequencing costs.

Method used

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  • Kit and method for detecting hypertension drug related gene polymorphism
  • Kit and method for detecting hypertension drug related gene polymorphism
  • Kit and method for detecting hypertension drug related gene polymorphism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Design of primers for polymorphic sites of genes associated with hypertension medication

[0054] (1) Determine the second-round PCR general primers P3 and P4, the specific sequences of which are shown in SEQ ID NO.361 and SEQ ID NO.362 in Table 1.

[0055] The "NNNNNNNN" fragment in P4 is Index, which is an oligonucleotide sequence composed of dATP, dTTP, dCTP, and dGTP arbitrarily.

[0056] (2) Design the first round of PCR upstream primer P1 and downstream primer P2; wherein P1 is composed of two parts P1-1 and P1-2, and P2 is composed of two parts P2-1 and P2-2.

[0057] P1-1 and P2-1 are the 22 bases at the ends of P3 and P4 in the 5' to 3' direction, respectively, and the specific sequences are shown in Table 3 below:

[0058] Table 3 Base sequences of P1-1 and P2-1

[0059] P1-1 CTACACGACGCTCTTCCGATCT P2-1 CAGACGTGTGCTCTTCCGATCT

[0060] Design P1-2 and P2-2, including steps:

[0061] Ⅰ. Determine the site sequence on NCBI;

[...

Embodiment 2

[0073] Embodiment 2 library construction

[0074] 1. Take three cases of clinical anticoagulant blood samples (sample numbers: 013256, 031638, 043219), use self-prepared or commercial nucleic acid extraction kits to extract human genomic DNA, and use NanoDrop 2000 ultra-micro-volume ultraviolet spectrophotometer to detect A260 / A280, Qubit quantitative sample concentration C (ng / μl). A260 / A280=1.7~1.9 The extraction results are shown in Table 5 below:

[0075] table 5

[0076] sample number 013256 031638 043219 C (ng / μl) 19.4 27.1 23.1 A260 / A280 1.82 1.81 1.76

[0077] 2. The first round of multiplex PCR reaction

[0078] Using the multiple primer working solution designed in Example 1 (which contains the primer set shown in SEQ ID NO.1 to SEQ ID NO.360 in Table 2) to perform the first round of amplification on the sample and blank control.

[0079] The amplification system is shown in Table 6 below:

[0080] Table 6 Amplification system ...

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Abstract

The invention discloses a primer group, a kit and a method for detecting hypertension drug related gene polymorphism. The primer group is selected from SEQ ID NO.1-SEQ ID NO.360 nucleotide sequences.The primer group can directly perform PCR after multiple PCR capturing; the primer group and the kit are simple to operate, and experiment steps and check are reduced; besides, consumable and labor cost for construction of a DNA library can be further reduced; drug kinds guided by gene sites corresponding to 180 pairs of primers comprise fix common hypertension treatment drugs and hypertension complication drugs including anti-freezing, anti-platelet, sugar-reducing, aldosterone and statin common drugs, selection of group treatment schemes for complications such as newly diagnosed hypertension, H type hypertension, refractory hypertension and hypertension atherosclerosis can be guided, so that patients can be effectively treated as early as possible.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a kit and a method for detecting gene polymorphisms related to hypertension medication. Background technique [0002] As the most important risk factor for cardiovascular and cerebrovascular diseases, hypertension is also a chronic disease with a high prevalence rate, high disability rate and heavy disease burden in my country. According to data released by the National Health and Family Planning Commission in 2016, the prevalence of hypertension in adults aged 18 and over in my country is 25.2%, and the epidemic situation is serious. Its main complications, such as stroke, myocardial infarction, heart failure and chronic kidney disease, have a high disability and mortality rate, seriously consume medical and social resources, and cause a heavy burden to families and society. It has become an important public health problem in my country. . [0003] Drug therapy is curren...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 夏昊强周煌凯刘倩王芳程祖福陆嫚云姚啟聪
Owner 广州海思医疗科技有限公司
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