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Primer and probe group for detecting rs4149056 and application thereof

A primer-probe, probe technology, applied in recombinant DNA technology, microbial assay/test, biochemical equipment and methods, etc., can solve the problems of increasing statin concentration, decreasing transport function, affecting treatment effect, etc. Significant application promotion value, easy operation effect

Inactive Publication Date: 2018-12-21
潍坊德诺泰克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the SLCO1B1 gene is mutated, its transport function will decrease, which will seriously affect the absorption of drugs by the liver, will lead to an increase in the concentration of statins in the blood, and increase the adverse reactions of drugs (such as rhabdomyolysis, etc.), which will affect the therapeutic effect

Method used

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  • Primer and probe group for detecting rs4149056 and application thereof
  • Primer and probe group for detecting rs4149056 and application thereof
  • Primer and probe group for detecting rs4149056 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1, be used to detect the design of the primer probe set of rs4149056

[0051] After a lot of design, pre-experiment, and effect comparison, a set of primer probes for detecting rs4149056 was obtained.

[0052] Primer F: 5'-AAAGGAATCTGGGTCATACATGTGGA-3';

[0053] Primer R: 5'-TTAGCGAAATCATCAATGTAAGAAAG-3';

[0054] Probe P: 5'-FAM- TGC GTTCATGGGTAATATGCTTC GCA -BHQ1-3'.

[0055] 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, 24, 25, and 26) are locked nucleic acids (LNA).

[0056] The target sequences of primer F and primer R are shown in ...

Embodiment 2

[0057] Embodiment 2, the application of the primer probe set for detecting rs4149056

[0058] 1. Blood sample processing (processing method for a single blood sample)

[0059] 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.

[0060] Lysis solution: 1.7g / 100ml NaCl aqueous solution.

[0061] 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.

[0062] 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 and probe group for detecting rs4149056 and application thereof. Firstly, the primer and probe group is protected, and consists of a primer F, a primer R and a probe P, wherein the primer F is a single-chain DNA (deoxyribonucleic acid) molecule shown in a sequence 1 of a sequence list; the primer R is a single-chain DNA molecule shown in a sequence 2 of the sequence list; the probe P is a single-chain DNA molecule of which one terminal has a fluorescence group and the other terminal has a fluorescence quenching group, one part of nucleotide in the DNA moleculeis locked nucleic acid, and a nucleotide sequence of the DNA molecule is shown in a sequence 3 of the sequence list. The primer and probe group can be used for detecting the rs4149056, and judging thegene type of the to-be-tested sample based on the site, so as to instruct medicine usage; the important application and popularization value is realized.

Description

technical field [0001] The invention relates to a primer probe set for detecting rs4149056 and application thereof. Background technique [0002] The SLCO1B1 gene encodes the organic anion transporting polypeptide OATP1B1. OATP1B1 is a transmembrane transporter that is mainly distributed in the liver. Glucose, rifampicin, angiotensin Ⅱ, receptor antagonists and other drugs play an important regulatory role. In recent years, multiple single nucleotide polymorphisms (SNPs) have been found on the SLC01B1 gene, some of which have been confirmed to be closely related to the transport function in vitro or in vivo. [0003] Tests have confirmed that the SLC01B1 521 T>C gene mutation will lead to a decrease in the non-renal clearance rate of the statin lipid-lowering drug pravastatin, and an increase in blood drug concentration. It has been reported that the frequency of 521 T>C in the Japanese population is about 15.8%, and in China The frequency of mutations in the populat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2531/113C12Q2525/301C12Q2525/307
Inventor 丛茜刘婷婷李朋赵海文张可谦孙辉管帅赵斌
Owner 潍坊德诺泰克生物科技有限公司
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