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Hypertension medication gene detection kit and use method thereof

A technology for gene detection and antihypertensive drugs, applied in the field of genetic detection kits for hypertension drugs, can solve the problems of high cost, complicated operation, and high sample processing requirements

Active Publication Date: 2020-06-26
重庆浦洛通基因医学研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims at detecting CYP2D6*10, CYP2C9*3, ADRB1, AGTR1, NPPA, CYP3A5*3 and ACE (I / D) type gene polymorphisms existing in the prior art, requiring high sample processing, long detection period, To solve the problems of complex operation, high cost, and low specificity, a genetic detection kit for hypertension medication and its use method are provided

Method used

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  • Hypertension medication gene detection kit and use method thereof
  • Hypertension medication gene detection kit and use method thereof
  • Hypertension medication gene detection kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1: Primer and probe combination design and use

[0086] 1. Primer and probe combination for identifying CYP2D6>C:

[0087] CYP2D-WR1:CAACGCTGGGCTGCACGCTACC

[0088] CYP2D-F1:TCCCTCACCTGGTCGAAGCAGT

[0089] CYP2D-P1:FAM-TCTGGAAGTCCACATGCAGCAGGTT–TRAMA

[0090] CYP2D-WB:CGCTGGGCTGCACGCTACCCACCAGGCCCCCTGCC-ddC

[0091] Primer and probe combinations used to identify CYP2D6>T:

[0092] CYP2D-MR1:CAACGCTGGGCTGCACGCTACT

[0093] CYP2D-F1:TCCCTCACCTGGTCGAAGCAGT

[0094] CYP2D-P1:FAM-TCTGGAAGTCCACATGCAGCAGGTT–TRAMA

[0095] CYP2D-MB:CGCTGGGCTGCACGCTACTCACCAGGCCCCCTGCC-ddC

[0096] The fluorescent group at the 5' end of the above probe is a routinely used fluorescent reporter group suitable for fluorescent quantitative PCR analysis, preferably FAM, VIC, HEX, cy5 or ROX, and the quencher group at the 3' end is suitable for fluorescent quantitative PCR. The conventionally used fluorescence quenching group is preferably TAMRA, BHQ1, BHQ2, MGB or Dabcy1, and a more pr...

Embodiment 2

[0164] Example 2: Acquisition of Positive Quality Controls

[0165] The acquisition method of the positive quality control product is: according to the CYP2D6, ADRB1, CYP2C9, AGTR1, ACE, NPPA, CYP3A57 gene sequences announced by the NCBI database, construct a synthetic sequence gene fragment of the plasmid, and then insert the fragment into the T vector, Escherichia coli DH5α strain was used to transform and extract plasmids, and the quality control plasmids were mixed in equal proportions to obtain positive quality control products.

[0166] The sequence of CYP2D6>C quality control substance is shown in SEQ ID NO.1, the sequence of CYP2D6>T quality control substance is shown in SEQ ID NO.2, the sequence of ADRB1>G quality control substance is shown in SEQ ID NO.3, and the sequence of ADRB1>G quality control substance is shown in SEQ ID NO.3. The sequence of C quality control product is shown in SEQ ID NO.4, the sequence of CYP2C9>A quality control product is shown in SEQ ID NO....

Embodiment 3

[0167] Embodiment 3: the configuration of PCR reaction solution

[0168] The kit of the present invention adopts the design of 8 PCR reaction strips, and tubes 1-7 are composed of gene detection reagents, respectively indicating CYP2D6*10, CYP2C9*3, ADRB1, AGTR1, NPPA, CYP3A5*3 and ACE (I / D) For gene polymorphic sites, tube No. 8 consists of detection reagents for the negative control. The reaction solutions in tubes 1-8 correspond to PCR reaction solutions 1-8 respectively, and the composition of PCR reaction solutions 1-8 is shown in Table 1-Table 8.

[0169] Table 1 PCR reaction solution 1

[0170]

[0171] Table 2 PCR reaction solution 2

[0172]

[0173]

[0174] Table 3 PCR reaction solution 3

[0175]

[0176] Table 4 PCR reaction solution 4

[0177]

[0178]

[0179] Table 5 PCR reaction solution 5

[0180]

[0181] Table 6 PCR reaction solution 6

[0182]

[0183]

[0184] Table 7 PCR reaction solution 7

[0185]

[0186] Table 8 P...

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Abstract

The present invention provides a hypertension medication gene detection kit and relates to an antihypertensive drug beta1 receptor blocker, an angiotensin II receptor antagonist, an angiotensin converting enzyme inhibitor, a diuretic and a calcium ion antagonist. The hypertension medication gene detection kit comprises primer pairs, probes and inhibitors for detecting CYP2D6*10, CYP2C9*3, ADRB1, AGTR1, NPPA, CYP3A5*3 and ACE (I / D) genes, a PCR reaction solution, positive quality control products and negative quality control products. The present invention also provides a use method of the hypertension medication gene detection kit. The hypertension medication gene detection kit solves problems of high sample processing requirements, long detection cycle, complicated operation, high cost, not strong specificity, etc. when detecting genetic polymorphism.

Description

technical field [0001] The invention is applied in the technical field of gene detection, and in particular relates to a genetic detection kit for hypertension medication and a use method thereof. Background technique [0002] The number of hypertensive patients in my country is as high as hundreds of millions, and the incidence rate is still on the rise. The complications caused by it have become the second leading cause of death in my country. Controlling blood pressure through drug therapy to reduce hypertensive complications has become the main treatment for hypertension. However, it is very common that there are individual differences in clinical responses to drugs for the treatment of hypertension, and 20-50% of patients receiving drug therapy have not well controlled blood pressure. The main reason is also due to genetic variation in drug-related drug-metabolizing enzymes and receptors. Individual differences in drug efficacy and adverse reactions are common phenome...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6883C12Q1/6858C12Q2600/106C12Q2600/156C12Q2531/113C12Q2563/107
Inventor 韩勋领李红玉罗锋
Owner 重庆浦洛通基因医学研究院有限公司
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