A set of primers for detecting SNP sites of drug metabolizing enzyme genes and its application
A technology of metabolic enzymes and loci, which is applied in the field of complete primers for the detection of SNP loci of drug metabolizing enzyme genes, can solve the problems of time-consuming, incomplete detection of genes, and complicated operations, so as to save cost and time, and improve irrational drug use The status quo, the effect of easy operation
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Embodiment 1
[0033] The establishment of embodiment 1 technical scheme
[0034] S1 design technical route: determine the main components of children’s common drugs (Table 1), clarify the results of SNP typing of genes and sites related to drug metabolism and the interpretation of drug metabolism rate (Table 2);
[0035] S2 establishes the reaction system: design multiple PCR amplification primers (Table 3) and extension primers (Table 4), adjust the concentration of reaction components and reaction conditions;
[0036] S2 Analyzing samples: DNA extraction, multiplex PCR amplification, time-of-flight mass spectrometry detection, result analysis and medication advice.
[0037] Table 1 Common drugs and their main ingredients for children
[0038]
[0039] Table 2 Types of related SNP sites and drug metabolism rate
[0040]
[0041] Table 3 multiplex PCR amplification primer sequence
[0042]
[0043] Table 4 Single base extension primer sequence
[0044] serial number ...
Embodiment 2
[0045] Example 2 sample extraction and analysis
[0046] In order to verify the detection accuracy and effectiveness of the set of primers used to detect drug-metabolizing enzyme genes of the present invention, 10 samples were selected for gene detection, and the process was as follows:
[0047] A. Saliva DNA extraction: use Kangwei Century Oral Swab Genomic DNA Extraction Kit for saliva DNA extraction. Add a certain amount of absolute ethanol to the GW1 and GW2 reagents. Take 500 μL saliva sample from the sampling tube, add 300 μL GR, 20 μL Proteinase K and 300 μL GL, shake and mix, incubate with shaking at 56°C for 15 minutes, add 300 μL absolute ethanol, shake and mix; add 750 μL of this solution to the adsorption tube of the collection tube Add 400 μL GW1 to the adsorption column, centrifuge at 12,000 rpm for 1 min, discard the liquid; add 400 μL GW2 to the adsorption column, centrifuge at 12,000 rpm for 1 min, discard the liquid; centrifuge at 12,000 rpm for 2 min, and let...
Embodiment 3
[0071] Example 3 Analysis of genetic testing results
[0072] The 9 SNP site gene detection results (Table 5) of the 10 samples in Example 2 were analyzed, and the rational drug use suggestions were given in conjunction with the drug-metabolizing enzyme gene-related SNP site typing and drug metabolism rate (Table 2) .
[0073] Table 5 Genotypes of 9 SNP loci in drug metabolism-related genes
[0074] site 1 2 3 4 5 6 7 8 9 10 rs1065852 A AG AG AG A G AG AG A AG rs762551 A A AC C AC AC AC A A A rs1057910 A A A A A A A A A A rs4244285 G G G AG AG G AG AG G G rs4986893 AG G G G G G AG G G G rs28371759 A A A A A A A A A A rs776746 C C CT C CT C C T CT C rs3813867 G G CG G G G CG G G G rs2031920 C C CT CT T C C C C CT
[0075] It can be seen from Table 5 that the genotypes of the 9 loci can be detected correctl...
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