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Primer combination, reagent kit and method for detecting CYP3A5*3

A technology for detection kits and detection primers, applied in biochemical equipment and methods, measurement/testing of microorganisms, recombinant DNA technology, etc., can solve the problems of high cost of CYP3A5 genotyping, improvement of detection efficiency, complicated operation procedures, etc. Achieve high yield, rapid amplification reaction and low cost

Inactive Publication Date: 2016-08-17
BEIJING JINQI BIOLOGICAL TECH CO LTD
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is that the existing CYP3A5 genotyping detection has problems such as high cost, complicated operation procedures, and long detection cycle, which is not conducive to improving detection efficiency and realizing clinical popularization

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  • Primer combination, reagent kit and method for detecting CYP3A5*3
  • Primer combination, reagent kit and method for detecting CYP3A5*3
  • Primer combination, reagent kit and method for detecting CYP3A5*3

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Embodiment Construction

[0050] In order to better understand the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings, but the protection scope of the present invention is not limited.

[0051] A detection primer composition for CYP3A5*3, comprising:

[0052] Forward outer primer F3:

[0053] 5'-TACCACCCAGCTTAACGA-3' (SEQ ID No. 1);

[0054] Reverse Outer Primer B3:

[0055] 5'-CCATACCCCTAGTTGTACG-3' (SEQ ID No. 2);

[0056] Forward inner primer FIP:

[0057] 5'-TCCAAACAGGGAAGAGATATTGAAAATGCTCTACTGTCATTTTCTAACC-3' (SEQ ID No. 3);

[0058] Reverse inner primer BIP:

[0059] 5'-GCTCCTGTGTGAGACTCTTGCACACACAGCAACCTTAGG-3' (SEQ ID No. 4); and

[0060] Peptide nucleic acid sequence PNA-A for blocking the rs776746A site:

[0061] 5'-CAATATCTCTTTCCCTGT-3' (SEQ ID No. 5);

[0062] The peptide nucleic acid sequence PNA-G used to block the rs776746G site:

[0063] 5'-AGTATCTCTTCCCTGTT-3' (SEQ ID No. 6);

[0064] Pep...

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Abstract

The invention relates to the technical field of biological detection, in particular to a primer combination, a reagent kit and a method for detecting CYP3A5*3. PNA (peptide nucleic acid) Clamp LAMP (loop-mediated isothermal amplification) technologies are adopted, PNA and LAMP processes are combined with one another, A sites of rs776746 are blocked by PNA-A, G sites of the rs776746 are blocked by PNA-G, highly similar genes CYP3A4 are blocked by PNA-I, accordingly, corresponding LAMP reaction cannot continue to be carried out, and the purpose of detecting CYP3A5 genotypes can be achieved. The primer combination, the reagent kit and the method have the advantages that the method includes simple steps, the primer combination, the reagent kit and the method are high in specificity and low in cost, and the CYP3A5*3 is easy and convenient to identify.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a CYP3A5*3 detection primer composition, kit and method. Background technique [0002] Cytochrome P450 (cytochrome P450 or CYP450) referred to as CYP450, mainly exists in the liver and intestinal monooxygenase, is a group of superfamily enzymes involved in the metabolism of endogenous substances and exogenous compounds in the human body, especially in the drug play an important role in the metabolic process in the body. Among them, CYP3A5 is involved in the metabolism of various drugs such as tacrolimus, midazolam, dapsone, cortisone, and nifedipine. A SNP of this gene, rs776746 (CYP3A5*3, 6986A>G), will cause the mRNA of CYP3A5 gene to be cut too short to form an "incomplete" CYP3A5 protein, which will lead to the reduction or even disappearance of CYP3A5 enzyme protein activity. According to statistics, the frequency of CYP3A5*3 alleles in the Chin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q2531/119C12Q2525/107
Inventor 汪大为张井存
Owner BEIJING JINQI BIOLOGICAL TECH CO LTD
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