Primer pair, kit and method for detection of related genes for guiding individualized administration of fentanyl drugs

A gene detection and kit technology, applied in the field of gene detection related to guiding the individualized medication of fentanyl drugs, can solve the problems of no kit products, adverse reactions of clinical medication, etc., and achieve lower medical costs, rapid detection, and specificity. Good results

Inactive Publication Date: 2018-04-10
韩林志
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the technical problems of traditional clinical experience medical diagnosis and fentanyl drug administration, which are prone to clinical adverse reactions and there is no suitable kit product, the present invention provides a simple operation, rapid detection, high accuracy and specificity Good primer pairs, kits and methods for guiding the detection of genes related to individualized drug use of fentanyl drugs

Method used

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  • Primer pair, kit and method for detection of related genes for guiding individualized administration of fentanyl drugs
  • Primer pair, kit and method for detection of related genes for guiding individualized administration of fentanyl drugs
  • Primer pair, kit and method for detection of related genes for guiding individualized administration of fentanyl drugs

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Experimental program
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Embodiment 1

[0036] Embodiment 1: the preparation of kit

[0037] 1. Design and synthesis of primers and probes

[0038]Aiming at the CYP3A4*1G gene locus in the human genome (for the sequence, please refer to the human whole genome sequence published in the NCBI database), specific primers and MGB-labeled probes were designed using Primer Premier 3.0 and Methyl Primer Express v1.0 software, respectively.

[0039] Specific primers and probe sequences are shown in the table below:

[0040]

[0041] 2. Selection of reference substance

[0042] Positive control substance one is the wild-type plasmid of CYP3A4*1G gene locus;

[0043] The second positive control substance is a plasmid heterozygous for the CYP3A4*1G gene locus wild type and mutant type at a quantitative ratio of 1:1;

[0044] Positive control product three is a CYP3A4*1G gene locus mutant plasmid;

[0045] The blank control substance was nuclease-free water.

[0046] 3. Composition of PCR reaction solution

[0047] Incl...

Embodiment 2

[0057] Example 2: Using the above-mentioned kit to perform a method for detecting genes related to individualized drug use of fentanyl drugs

[0058] 1. Biological samples

[0059] The biological materials used in the present invention are all from Xiangya Medical Laboratory. Anticoagulation was performed for the remaining 500 cases tested at Xiangya Medical Laboratory during March-September 2017.

[0060] 2. Take the DNA extracted from the sample to be tested as a PCR template:

[0061] The DNA extraction kit is the self-developed extraction kit "Nucleic Acid Extraction or Purification Reagent" for extraction (record number: Xiangchang Machinery Equipment 20150166).

[0062] 1) Take a sterile 1.5mL EP tube and add 250uL whole blood.

[0063] 2) Add 750ul of cell lysate, mix by inverting 5-6 times, and let stand at room temperature for 10min.

[0064] 3) Centrifuge at 12000rpm for 1.5min; pour off the upper waste liquid and collect the sediment at the bottom of the tube. ...

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Abstract

The invention relates to a primer pair for the detection of related genes for guiding the individualized administration of fentanyl drugs. The primer pair comprises specific primers and a probe aimingat a CYP3A4*1G gene site. The invention relates to a kit for the detection of related genes for guiding the individualized administration of fentanyl drugs. The kit comprises a PCR reaction solutioncontaining the primers and the probe mentioned above, PCR buffer, dNTP, nuclease-free water, and HS Tag enzymes. The invention also relates to a method for the detection of related genes for guiding the individualized administration of fentanyl drugs. The provided kit and detection method thereof have the advantages that the operation is simple, the detection is rapid, the accuracy is high, the specificity and sensitivity are good, the use is convenient, and the clinical requirements can be satisfied effectively.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a primer pair, a kit and a method for guiding the detection of genes related to individualized medication of fentanyl drugs. Background technique [0002] The CYP3A family is the drug-metabolizing enzyme that mediates the largest amount of drug metabolism in the human body. It accounts for 70% of the total CYP450 in the body and 30% of the CYP450 in the liver, and participates in the metabolic transformation of more than 60% of commonly used drugs in the body. At present, four CYP3A~ genes have been found in the human body, namely CYP3A4, CYF3A5, CYP3A7 and CYP3A43, and CYP3A4 is one of the most clinically important P450 enzymes, involved in the oxidative metabolism of more than 150 drugs in 38 categories, Accounting for about 50% of all drugs, it is the root cause of adverse reactions of many drugs due to interactions. [0003] There are obvious individual difference...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/6883C12Q1/686C12Q2600/106C12Q2600/156C12Q2563/107C12Q2561/101
Inventor 韩林志肖芳邓思铭
Owner 韩林志
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