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Primer-probe group for detecting rs 1057910 and application thereof

A primer probe and probe technology, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems of decreased ability to oxidize and metabolize drugs in the population, increased plasma drug concentration, poisoning, etc. Achieve the effect of significant application and promotion value and easy operation

Inactive Publication Date: 2018-12-04
山东德诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ability of people carrying CYP2C9*3 gene polymorphisms to oxidize and metabolize drugs is greatly reduced, resulting in a significant increase in plasma drug concentrations in vivo
Among these drugs, the metabolism of some drugs with a relatively narrow therapeutic index, such as warfarin, tolbutamide, and phenytoin, has received more attention, because the impairment of CYP2C9 metabolic activity may affect the actual dose of the drug in vivo, and May cause poisoning

Method used

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  • Primer-probe group for detecting rs 1057910 and application thereof
  • Primer-probe group for detecting rs 1057910 and application thereof
  • Primer-probe group for detecting rs 1057910 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1, be used to detect the design of the primer probe group of rs1057910

[0052] After a lot of design, pre-experiment, and effect comparison, a set of primer probes for detecting rs1057910 was obtained.

[0053] Primer F: 5'-GAGCCACATGCCCTACACAGAT-3';

[0054] Primer R: 5'-CTTACCTTGGGAATGAGATAGTTT-3';

[0055] Probe P: 5'-FAM- ACA GGTGGGGAGAAGGTCAA TGT -BHQ1-3'.

[0056] Primer F is a single-stranded DNA molecule shown in Sequence 1 of the Sequence Listing. Primer R is a single-stranded DNA molecule shown in Sequence 2 of the Sequence Listing. Probe P is a single-stranded DNA molecule with a FAM fluorescent group at the 5' end and a BHQ1 fluorescent quencher group at the 3' end. The nucleotide sequence of the DNA molecule is shown in sequence 3 in the sequence table, and the underlined nucleosides Acids (i.e., positions 1, 2, 3, 21, 22, and 23) are locked nucleic acids (LNA).

[0057] The target sequences of primer F and primer R are shown in sequenc...

Embodiment 2

[0058] Embodiment 2, the application of the primer probe set for detecting rs1057910

[0059] 1. Blood sample processing (processing method for a single blood sample)

[0060] 1. Take 65 μL of EDTA anticoagulant blood, add 500 μL of lysate, mix by inversion, centrifuge at 12000 rpm for 4 min, and discard the supernatant.

[0061] Lysis solution: 1.7g / 100ml NaCl aqueous solution.

[0062] 2. After completing step 1, add 120 μL of preservation solution, bathe in water at 65°C for 10 minutes, then in water at 85°C for 10 minutes, then centrifuge at 12,000 rpm for 4 minutes, and store at 4°C for later use. Take the supernatant before use, which is the template sample.

[0063] The preparation method of the preservation solution: ① mix 25 parts by volume of FG buffer solution and 4 parts by volume of proteinase K solution to obtain a mixed solution; ② mix 1 part by volume of the mixed solution obtained in step ① with 5 parts by volume of ddH 2 O mixed to obtain a preservation sol...

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Abstract

The invention discloses a primer-probe group for detecting rs 1057910 and an application thereof and firstly, protects the primer-probe group composed of a primer F, a primer R and a probe P, whereinthe primer F is a single-chain DNA molecule represented as the SEQ No.1 in the sequence table, the primer R is a single-chain DNA molecule represented as the SEQ No.2 in the sequence table, and the probe P is a single-chain DNA molecule, of which one terminal has a fluorophore while the other terminal has a fluorescence quenching group, a part of nucleotides in the DNA molecule being locked nucleic acid; the nucleotide sequence of the DNA molecule in the probe P is represented as the SEQ No.3 in the sequence table. The primer-probe group can be used for detecting rs 1057910 in order to determine the genotype of a to-be-detected sample on the basis of the loca, thereby guiding use of medicines. The primer-probe group has important application and promotion values.

Description

technical field [0001] The invention relates to a primer probe set for detecting rs1057910 and application thereof. Background technique [0002] Cytochrome P450 (Cytochrome P450 or CYP450, referred to as CYP450) represents a large family of heme proteins that can be oxidized by itself, and belongs to the class of monooxygenases. It is named for its specific absorption peak at 450 nanometers. It is involved in the metabolism of endogenous substances and exogenous substances including drugs, environmental compounds. According to the degree of amino acid sequence homology, its members are divided into three levels: family, subfamily and enzyme individual. The cytochrome P450 enzyme system can be abbreviated as CYP, where the family is represented by Arabic numerals, the subfamily is represented by uppercase English letters, and the individual enzymes are represented by Arabic numerals, such as CYP2D6, CYP2C19, CYP3A4, etc. At least 9 P450s in human hepatic cytochrome P450 en...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/106C12Q2600/156C12Q2563/107C12Q2545/114
Inventor 丛茜赵海文刘婷婷李朋孙文文王洪霞李晓妮任妮妮
Owner 山东德诺生物科技有限公司
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