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Gene polymorphism detection kit for salbutamol metabolism marker and detection method and use thereof

A technology of genetic polymorphism and metabolic markers, applied in the field of genetic testing, can solve the problems of cumbersome operation and time-consuming

Inactive Publication Date: 2021-12-03
上海普然生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the methods of obtaining blood DNA mainly include traditional column extraction and magnetic bead extraction, both of which are time-consuming and cumbersome to operate

Method used

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  • Gene polymorphism detection kit for salbutamol metabolism marker and detection method and use thereof
  • Gene polymorphism detection kit for salbutamol metabolism marker and detection method and use thereof
  • Gene polymorphism detection kit for salbutamol metabolism marker and detection method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the preparation of kit

[0042] (1) Specific primer design

[0043] The kit of the present invention designs specific amplification primers and sequencing primers for the ADRB2 (A46G) gene polymorphism, and is used for pyrophosphate PCR detection. The gene polymorphism sequence is subject to the public sequence in Genebank, and the primer sequences are shown in the table below.

[0044]

[0045] (2) Kit composition

[0046] The detection kit includes the components shown in Table 2 below:

[0047] Table 2 Kit component list

[0048]

[0049]

[0050] (3) The configuration system of the sample treatment solution is as follows:

[0051] Sample treatment solution, including 0.4% lithium dodecyl sulfate, 0.3% Triton X-100, 15mg / mL sodium hydroxide, 10mM betaine, 10% trehalose, 5mM BSA, 50mM Tris-HCl, 100mM NaCl, pH=9.

[0052] (4) The detection kit amplification reagent 1 single serving configuration system of the present embodiment is as follow...

Embodiment 2

[0059] Embodiment 2, kit detection steps

[0060] The instruments adopted in the present invention are as follows: thermostat, pyrosequencer (Wuhan First Biotechnology Co., Ltd.).

[0061] 1) Take 30 μL EDTA anticoagulated whole blood sample in a PCR amplification tube;

[0062] 2) Add 100 μL of sample treatment solution and 4ul of magnetic beads, and let stand for 5 minutes;

[0063] 3) Place the PCR amplification tube on the magnetic stand, and suck out the mixed solution from the opposite side of the magnetic beads after all the magnetic beads are adsorbed to one side;

[0064] 4) Add amplification reagent 1 into the dry powder of amplification reagent 2, fully dissolve and mix;

[0065] 5) Add 25ul of the prepared PCR reaction solution into the PCR amplification tube obtained in step 4), fully mix the magnetic beads and the PCR reaction solution, centrifuge, and perform constant temperature reaction.

[0066] 6) Amplify using a PCR instrument, the reaction system is 25 ...

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Abstract

The invention discloses a gene polymorphism detection kit for a salbutamol metabolism marker and a detection method and use thereof. The salbutamol metabolism marker is a gene polymorphism of ADRB2A46G and a specific amplification primer and a sequencing primer aiming at the gene polymorphism of the ADRB2A46G are designed in the kit. The kit comprises the following components: a sample treating fluid, magnetic beads, an amplification reagent 1, an amplification reagent 2, an ADRB2A46G sequencing primer and a positive control. A mode of extracting and amplifying in a same tube is used, such that repeated tube transfer and a risk of nucleic acid loss during extraction are avoided. Rapid isothermal amplification is carried out by adding an anti-inhibitor. The kit aims to detect the gene polymorphism of a curative effect of salbutamol by combining constant-temperature PCR and pyrophosphoric acid detection, and provides suggestions from a gene perspective for clinical personalized medication.

Description

technical field [0001] The invention relates to a gene polymorphism detection kit for salbutamol metabolic markers, a detection method and an application thereof, and belongs to the field of gene detection. Background technique [0002] Salbutamol, a short-acting β2 adrenergic receptor agonist, is used as an antiasthmatic drug, which can effectively inhibit the release of allergenic substances such as histamine and prevent bronchospasm. Adding a small amount of salbutamol to livestock feed can increase the lean meat mass and meat replacement rate of livestock, and reduce fat, but its toxicity is much higher than that of ractopamine, which has the same function. It is suitable for bronchial asthma, asthmatic bronchitis, bronchospasm, emphysema and other diseases. Some patients have unsatisfactory effects after treatment with albuterol, and patients still have acute attacks. The different responses of patients to drug treatment cannot be simply considered to be caused by diff...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/686C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/686C12Q1/6869C12Q2600/156C12Q2600/106C12Q2523/308C12Q2563/143C12Q2563/149C12Q2565/301
Inventor 孙悦刘丹周虹桥
Owner 上海普然生物科技有限公司