Solid-phase PCR (Polymerase Chain Reaction) kit for gene detection of precision drug utilization of cardiovascular and cerebrovascular diseases

A technology for cardiovascular and cerebrovascular diseases and precise drug use, applied in the field of PCR application, can solve the problems of hindering the expansion of thrombus and no thrombolytic effect, and achieve the effects of low cost of reagent consumables, improved detection sensitivity and specificity, and simple operation.

Inactive Publication Date: 2018-05-25
安徽安龙基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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Method used

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  • Solid-phase PCR (Polymerase Chain Reaction) kit for gene detection of precision drug utilization of cardiovascular and cerebrovascular diseases
  • Solid-phase PCR (Polymerase Chain Reaction) kit for gene detection of precision drug utilization of cardiovascular and cerebrovascular diseases
  • Solid-phase PCR (Polymerase Chain Reaction) kit for gene detection of precision drug utilization of cardiovascular and cerebrovascular diseases

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0023] Example 1 Composition of a solid-phase PCR detection kit for precise medication of 13 types of cardiovascular and cerebrovascular diseases

[0024] The kit consists of a PCR reaction mixture and a 12-row tube or a 96-well PCR reaction plate. The PCR reaction mixture is 5×PCRBuffer (Tris-HCl pH8.5 100mM, KCl 500nM, MgCl 2 15nM, dNTPs10mM), Mg 2+ , 20 pairs of primers and hot-start DNA polymerase.

[0025] The kit is to seal a layer of paraffin mixture on the surface of the PCR reaction mixture, and the paraffin mixture is made by mixing solid paraffin and liquid paraffin according to 1:8 (W / V).

[0026] The reaction system (25ul) of the kit is: 23ul PCR reaction solution and 2ul template.

Example Embodiment

[0027] Example 2 Operation and result determination of the kit

[0028] 1. Extraction of genomic DNA

[0029] A buccal swab was used to extract the exfoliated cells on the inner surface of the human oral cavity, and the buccal swab genomic DNA extraction kit (Tiangen Biochemical Technology, item number: DP322) was used to extract DNA according to the kit instructions.

[0030] 2. Preparation of reaction system

[0031] Take the cardio-cerebrovascular disease precise drug gene detection solid-phase PCR reaction kit 12 strip tube or 96-well PCR reaction plate, and add 2ul of the extracted DNA.

[0032] 3. PCR amplification

[0033] Put the PCR tube into an ordinary PCR machine and follow the instructions in the instructions before proceeding with PCR amplification.

[0034] 4. Analysis of PCR products by 1.0% agarose gel electrophoresis

[0035] 5. Interpretation of results

[0036] The interpretation criteria for the results of this kit are as follows: Table 1

[0037] Detection target

Example Embodiment

[0038] Example 3 Interpretation of sequencing results and medication guidance

[0039] Using the DNA extracted from the oral swabs of patients with cardio-cerebrovascular diseases in Example 1 as a template, after amplification with the above kit, the following sequencing results are obtained. According to the sequencing results, the following guidelines for the application of drugs are given:

[0040]

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Abstract

The invention discloses a solid-phase PCR (Polymerase Chain Reaction) kit for gene detection of precision drug utilization of cardiovascular and cerebrovascular diseases. The solid-phase PCR kit comprises a PCR solution mixed solution and 12 parallel-arrayed pipes or a 96-pore PCR plate, wherein the PCR solution mixed solution is prepared from a primer set, hot-start DNA (Deoxyribonucleic Acid) polymerase and a PCR buffering solution. Compared with the prior art, the solid-phase PCR kit disclosed by the invention has the advantages that 1) the operation is simple and convenient; only a nucleicacid template needs to be added into a solid-phase PCR plate and then the solid-phase PCR plate can be directly put on a machine for amplification; the solid-phase PCR kit is simple to operate and good in clinical detection specificity; the sensitivity is equivalent to quantitative PCR, and the kit has relatively strong clinical application and popularization value; 2) the sensitivity is high andthe specificity is strong; the solid-phase PCR kit disclosed by the invention adopts the hot-start DNA polymerase so that non-specific amplification is avoided, and the sensitivity and the specificity of the detection are improved; 3) detection sites are complete and the kit is suitable for safety drug utilization guidance of the cardiovascular and cerebrovascular diseases. In conclusion, a specific primer sequence is adopted and the reliability of a detection result is ensured; a detection method has the advantages of simplicity in operation, time saving and labor saving, large detection flux and low cost of reagent consumable items.

Description

technical field [0001] The invention relates to the technical field of PCR application, in particular to a solid-phase PCR kit for gene detection of precise medication for cardiovascular and cerebrovascular diseases. Background technique [0002] In my country, 3.5 million people die from cardiovascular and cerebrovascular diseases every year, and an average of 1 person dies from cardiovascular and cerebrovascular diseases every 10 seconds. It can be seen that the prevention and treatment of cardiovascular and cerebrovascular diseases has become the top priority of safeguarding human health. Because each person's genetic reasons, gender, height and weight, environment, lifestyle, etc. are not the same, resulting in large differences in treatment effects. Now clinicians follow the treatment guidelines in principle, but the guidelines are aimed at groups, and there are great individual differences. Precision medicine formulates individualized treatment plans through genetic ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 韦玉军李航刘琳琳苏军
Owner 安徽安龙基因科技有限公司
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