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
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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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