CN108823301A - It is a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism - Google Patents
It is a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism Download PDFInfo
- Publication number
- CN108823301A CN108823301A CN201810660244.0A CN201810660244A CN108823301A CN 108823301 A CN108823301 A CN 108823301A CN 201810660244 A CN201810660244 A CN 201810660244A CN 108823301 A CN108823301 A CN 108823301A
- Authority
- CN
- China
- Prior art keywords
- dna
- homo sapiens
- sixty
- detection kit
- base extension
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6858—Allele-specific amplification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/16—Primer sets for multiplex assays
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism, the 66 PCR amplification primer pairs reacted in same reaction system including being designed according to VKORC1, UGT1A1, HLA-A, HLA-B, TPMT, NAT1, NAT2, G6PD, F5, F2, IFNL3, APOE, COMT, CYP1A2, CYP2B6, CYP2C19, CYP2C9, CYP2D6, CYP3A5, MTFHR, SLCO1B1, DPYD gene polynorphisms site and the 66 Single base extension primers reacted in same reaction system.Multiple PCR detection kit of the invention can quickly, specifically, efficiently detect the polymorphism of above-mentioned 85 SNP sites of 22 genes, so as to predict cardiovascular and cerebrovascular disease standard scheme curative effect, therapeutic scheme is targetedly formulated convenient for clinician, the other patient of different shaped is taken and treats with a certain discrimination and treats, to improve the treatment level of entire cardiovascular and cerebrovascular disease.
Description
Technical Field
The invention belongs to the technical field of biomedical molecular detection, and particularly relates to a multiple PCR detection kit for detecting human drug gene polymorphism.
Background
The genetic variation of the genes of the in vivo metabolism, transport and action targets of the drugs and the change of the expression level thereof can cause drug-responsive individual differences by influencing the in vivo concentration and sensitivity of the drugs. In recent years, with the development of human genomics, the field of pharmacogenomics has been rapidly developed, and more drug genome biomarkers and detection methods thereof emerge successively. Pharmacogenomics has become an important tool for guiding clinical individualized medication, evaluating the risk of serious adverse drug reactions, guiding the research and development of new drugs and evaluating new drugs, and part of new drugs on the market are limited to specific genotype indication patients. The drug genome information is approved to be added in drug labels of more than 140 drugs by the United states FDa, and 42 drug genome biomarkers are involved. In addition, part of the industry guidelines also includes the detection of some non-FDa approved biomarkers and their properties (e.g., MgMt gene methylation) as guidelines for the treatment of disease. Molecular detection of drug response-related genes and their expression products is a prerequisite for the implementation of personalized drug therapy.
The key link between pharmacology and genetics includes both Pharmacokinetic (PK) and Pharmacodynamic (PD) aspects. Pharmacokinetics mainly aims at quantitatively researching absorption, distribution, metabolism and excretion rules of a medicament in an organism and emphasizes on clarifying the in-vivo process of the medicament; the pharmacokinetics mainly studies the action, action rule and action mechanism of the drug on the body, and the content thereof includes biochemical, physiological and morphological changes caused by the interaction between the drug and the action target, and focuses on explaining how the drug acts on the action target. The detection of drug metabolizing enzyme and drug target gene can guide the clinical selection of proper drug and administration dosage for specific patients, and realize individualized administration, thereby improving the effectiveness and safety of drug treatment and preventing the occurrence of serious adverse drug reactions.
The guiding of personalized medicine according to the detection of the drug genome biomarkers mainly comprises two types: firstly, the dosage of the medicine is adjusted according to the genetic information of an individual so as to increase the curative effect of the medicine and reduce the occurrence of adverse reaction of the medicine; secondly, the type of the medicine is determined according to the genetic information of the individual, and the medicine which is ineffective or possibly generates serious adverse drug reactions aiming at the specific genotype individual is avoided. Adjustment of drug dosage is often based on the results of randomized controlled clinical studies; for genetic variation lacking random control clinical research at present, the dosage of the drug can be estimated according to the influence of the genotype on the area under the pharmacokinetic curve of the drug; when the reactivity of a drug is influenced by a plurality of genes or the interaction between the genes and environmental factors, the dosage can be determined according to a dosage calculation formula which is deduced in large-scale clinical tests at home and abroad and contains the individual genotype and other factors. The guide suggestion of the genetic variation detection result of the common drug metabolizing enzyme and the drug action target point gene to clinical medication.
Disclosure of Invention
The invention aims to provide a multiplex PCR detection kit for detecting human drug gene polymorphism.
The detection principle of the invention is as follows:
the multiplex PCR detection kit disclosed by the invention is used for respectively designing and synthesizing a PCR amplification specific primer pair and a single base extension primer aiming at 66 sites, and then carrying out PCR amplification, SAP reaction and iPLEX single base extension reaction. Sample single base extension products are desalted through resin, then molecular weight difference among different extension products is detected through analysis of a MalDI-tOF (matrix-assisted laser desorption ionization-time of flight) mass spectrometer, base types of the 85 SNP sites are determined, genotyping is carried out, the genotypes can be directly judged, and different treatment modes are adopted according to patients with different genotyping.
The technical scheme of the invention is as follows:
a multiplex PCR detection kit for detecting human drug gene polymorphism comprises a sixty-six PCR amplification primer pair which is designed according to polymorphic sites of VKORC1, UGT1A1, HLA-A, HLA-B, TPMT, NAT1, NAT2, G6PD, F5, F2, IFNL3, APOE, COMT, CYP1A2, CYP2B6, CYP2C19, CYP2C9, CYP2D6, CYP3A5, MTFHR, SLCO1B1 and DPYD genes and reacts in the same reaction system and a sixty-six single-base extension primer, wherein,
the sixty-six PCR amplification primer pairs comprise first to sixty-six amplification primer pairs, wherein forward primers of the first to sixty-six amplification primer pairs are sequentially shown as SEQ ID NO 01-066, and reverse primers are sequentially shown as SEQ ID NO 067-132; the sixteenth single-base extension primer comprises first to sixteenth single-base extension primers which are sequentially shown as SEQ ID NOS 133-198;
the using method comprises the following steps:
(1) taking the genomic DNA to be detected as a template, and carrying out PCR amplification reaction in a PCR reaction system containing a RingCap-Taq enzyme and a RingCap buffer by using the sixty-six PCR amplification primer pair to obtain a PCR amplification reaction solution;
(2) carrying out SAP reaction on the PCR amplification reaction solution and SAP enzyme to obtain SAP reaction solution;
(3) performing single-base extension reaction on the SAP reaction solution by using the sixty-six single-base extension primer pair;
(4) and (4) desalting the material obtained in the step (3) by using resin, loading the material on a SpectrocHIP chip, and performing mass spectrometry detection to obtain the material.
In a preferred embodiment of the present invention, the conditions of the PCR amplification reaction include: 3min at 95 ℃ for 1 cycle; 45 cycles of 95 ℃ for 20s, 60 ℃ for 30s, 72 ℃ for 20 s; 3min at 72 ℃ for 1 cycle.
Further preferably, the conditions under which the SAP reacts include: 40min at 37 ℃, 5min at 85 ℃ and infinity at 4 ℃.
Still more preferably, the conditions for the single base extension reaction include:
the first stage consists of 40 cycles: each cycle comprises:
30s at 94 ℃, 1 cycle, 5s at 94 ℃, 5s at 52 ℃, 5s at 80 ℃, 5 cycles;
the second stage is 1 cycle in total: 3min at 72 ℃.
In a preferred embodiment of the invention, the RingCap-Taq enzyme consists of Taq enzyme, DNA ligase and DNA end modifying enzyme in a ratio of 0.8-1.2: 0.8-1.2.
Further preferably, the RingCap-Taq enzyme consists of Taq enzyme, DNA ligase and DNA end modifying enzyme in a ratio of 1: 1.
The invention has the beneficial effects that:
1. the multiplex PCR detection kit can quickly, specifically and efficiently detect the polymorphism of the 85 SNP sites of the 22 gene, thereby predicting the curative effect of a standard scheme of cardiovascular and cerebrovascular diseases, being convenient for clinicians to specifically formulate treatment schemes, and taking different treatment and treatment to patients of different types, thereby improving the diagnosis and treatment level of the whole cardiovascular and cerebrovascular diseases;
2. the PCR amplification primer, the single-base extension primer, the detection kit and the detection method for detecting 85 polymorphisms of the human 22 gene provided by the invention have high clinical application value and are particularly suitable for clinical popularization.
Drawings
FIG. 1 is a graph showing the result of mass spectrometry of the RS9923231 site in the 22 gene detected in example 2 of the present invention.
FIG. 2 is a graph showing the results of mass spectrometry of the RS776746 site in the 22 gene detected in example 2 of the present invention.
Detailed Description
The technical solution of the present invention will be further illustrated and described below with reference to the accompanying drawings by means of specific embodiments.
Example 1
A multiplex PCR detection kit for detecting human drug gene polymorphism comprises a sixty-six PCR amplification primer pair which is designed according to polymorphic sites of VKORC1, UGT1A1, HLA-A, HLA-B, TPMT, NAT1, NAT2, G6PD, F5, F2, IFNL3, APOE, COMT, CYP1A2, CYP2B6, CYP2C19, CYP2C9, CYP2D6, CYP3A5, MTFHR, SLCO1B1 and DPYD genes and reacts in the same reaction system, and a sixty-six single-base extension primer which reacts in the same reaction system;
the sixty-six PCR amplification primer pairs sequentially comprise first to sixty-six amplification primer pairs, forward primers of the first to sixty-six amplification primer pairs are sequentially shown as SEQ ID NO 01-066, reverse primers are sequentially shown as SEQ D NO 067-132, and the specific sequence is shown in the following table:
the sixty-six single-base extension primers sequentially comprise first to sixteenth single-base extension primers which are sequentially shown as SEQ ID NO 133-198, and are specifically shown in the following table:
the using method comprises the following steps:
(1) taking the genomic DNA to be detected as a template, and carrying out PCR amplification reaction in a PCR reaction system containing a RingCap-Taq enzyme and a RingCap buffer by using the sixty-six PCR amplification primer pair to obtain a PCR amplification reaction solution; the conditions of the PCR amplification reaction include: 3min at 95 ℃ for 1 cycle; 45 cycles of 95 ℃ for 20s, 60 ℃ for 30s, 72 ℃ for 20 s; 3min at 72 ℃ for 1 cycle; the RingCap-Taq enzyme consists of Taq enzyme, DNA ligase and DNA end modification enzyme in a ratio of 0.8-1.2: 0.8-1.2, preferably in a ratio of 1: 1;
(2) carrying out SAP reaction on the PCR amplification reaction solution and SAP enzyme to obtain SAP reaction solution; the conditions under which the SAP reacts include: 37 ℃ for 40min, 85 ℃ for 5min, and 4 ℃ infinity;
(3) performing single-base extension reaction on the SAP reaction solution by using the sixty-six single-base extension primer pair; the conditions of the single base extension reaction include:
the first stage consists of 40 cycles: each cycle comprises:
30s at 94 ℃, 1 cycle, 5s at 94 ℃, 5s at 52 ℃, 5s at 80 ℃, 5 cycles;
the second stage is 1 cycle in total: 72 ℃ for 3min
(4) And (4) desalting the material obtained in the step (3) by using resin, loading the material on a SpectrocHIP chip, and performing mass spectrometry detection to obtain the material.
In particular, the main reagents in the kit according to the invention are shown in the following table:
example 2 detection example of the multiplex PCR detection kit of example 1
Test sample: blood samples anticoagulated with EDta were obtained from the tumor hospital department of Fujian province and from healthy adults on routine physical examination.
Main instrument equipment：
1. DR massarrray mass spectrometry gene analysis system (guangzhou dary biotechnology limited);
2. aB-7500 fluorescent quantitative PCR instrument (applied biosystems, USA, aBI);
3. centrifuge (eppunerf);
4. rotary shaker (shanghai).
Detection program：
1. Purification of genomic DNA of blood samples: extracting with blood genome extraction kit.
2. And (3) PCR amplification: the purified DNA of the blood sample is used as a template, PCR amplification is carried out by using a PCR amplification primer pair aiming at 85 site polymorphisms in the kit provided in the embodiment 1, the reaction system is as follows,
；
a set of negative controls with water as the sample was set up.
The reaction conditions were as follows:
。
3. SAP reaction: and respectively adding 2 mul of SAP reaction solution into the 5 mul of PCR amplification reaction solution to perform SAP reaction, wherein the formula of the SAP reaction solution is as follows:
；
the reaction conditions were as follows:
4. single base extension reaction: adding 2 mul of extension reaction solution into the SAP digestive juice respectively to carry out single-base extension reaction, wherein the extension reaction solution formula comprises the following components:
(ii) a Wherein the single base extension primer mix is a mixture of single base extension primers for detecting 85 site polymorphisms in the kit provided in example 1.
The reaction conditions were as follows:
5. resin desalting: 41ul of water was added to each well of the extended reaction solution containing the sample and then centrifuged. Add 15mg of clean Resin (Resin): the sample plate is gently inverted in a row, placed on the sample plate with the resin, and then the sample plate is inverted (the two plates cannot move horizontally during the process) to allow the resin to fall into the wells. The plate was sealed with a membrane and placed on a rotator and shaken for 15 min. Plates were centrifuged for 5min at 3200g (4000 rpm of standard plate centrifuge).
6. SpectrocHIP chip loading: using MassarraYTMThe RS 1000 autosampler performs a Volume test (Volume check) to find the appropriate speed of application (dispensing speed). Suitable volumes for spotting are 8-12 nL. + -. 25%. Volume check with samples on plate, then MassarrayTMThe RS 1000 autosampler spotted the reacted sample onto a 96-spot spectrohip (chip).
7. And (4) detecting by mass spectrometry.
The detection results are shown in fig. 1 and fig. 2, and are compared with the sequencing results of two sites detected by the traditional gene sequencing method, so that the detection results of the two detection methods are proved to be completely identical, and the reliability of the detection method is proved.
The above description is only a preferred embodiment of the present invention, and therefore should not be taken as limiting the scope of the invention, which is defined by the appended claims.
Sequence listing
<110> Xiamen Biotech Co., Ltd
<120> a multiplex PCR detection kit for detecting human drug gene polymorphism
<160>198
<170>SIPOSequenceListing 1.0
<210>1
<211>21
<212>DNA
<213>Homo sapiens
<400>1
atgttggcca ggcttgtctt a 21
<210>2
<211>25
<212>DNA
<213>Homo sapiens
<400>2
gggacatgaa atagttgtcc tagca 25
<210>3
<211>21
<212>DNA
<213>Homo sapiens
<400>3
gttcctgccc ttccctttgt g 21
<210>4
<211>27
<212>DNA
<213>Homo sapiens
<400>4
tgatgagccc ccgtttaatc tattttt 27
<210>5
<211>24
<212>DNA
<213>Homo sapiens
<400>5
gaactcctcc taccctcatt gtgt 24
<210>6
<211>25
<212>DNA
<213>Homo sapiens
<400>6
atttccacac caactacact gtgtc 25
<210>7
<211>32
<212>DNA
<213>Homo sapiens
<400>7
ttttaaaaat tcatccatta cattttcagg ct 32
<210>8
<211>28
<212>DNA
<213>Homo sapiens
<400>8
gaagtctaag ctgattttct agaaccca 28
<210>9
<211>24
<212>DNA
<213>Homo sapiens
<400>9
gcatcaacct tctcaagaca acgt 24
<210>10
<211>22
<212>DNA
<213>Homo sapiens
<400>10
aggtgcaagg tagaagacac tg 22
<210>11
<211>22
<212>DNA
<213>Homo sapiens
<400>11
gtgaccattg atggcaggaa ct 22
<210>12
<211>30
<212>DNA
<213>Homo sapiens
<400>12
gacttggaaa cattaactga cattcttcaa 30
<210>13
<211>22
<212>DNA
<213>Homo sapiens
<400>13
ttcaccctca cccataggag at 22
<210>14
<211>24
<212>DNA
<213>Homo sapiens
<400>14
tcagaccaca atgttaggag ggta 24
<210>15
<211>26
<212>DNA
<213>Homo sapiens
<400>15
tgacagaaga gagaggaatc tggtac 26
<210>16
<211>28
<212>DNA
<213>Homo sapiens
<400>16
ggcttcatcc tcacctatag aaaattca 28
<210>17
<211>30
<212>DNA
<213>Homo sapiens
<400>17
gacttggaaa cattaactga cattcttgag 30
<210>18
<211>23
<212>DNA
<213>Homo sapiens
<400>18
ctgacaccac ccaccttgaa gaa 23
<210>19
<211>22
<212>DNA
<213>Homo sapiens
<400>19
gcaagcctta catctggctc at 22
<210>20
<211>22
<212>DNA
<213>Homo sapiens
<400>20
ccggtctgat agctcagaca ct 22
<210>21
<211>22
<212>DNA
<213>Homo sapiens
<400>21
ctcgaaggca tcacctacca tc 22
<210>22
<211>22
<212>DNA
<213>Homo sapiens
<400>22
gtagtggtcg atgcggtaga tc 22
<210>23
<211>28
<212>DNA
<213>Homo sapiens
<400>23
caattactgt tctcttgaag gaaatgcc 28
<210>24
<211>23
<212>DNA
<213>Homo sapiens
<400>24
ctggaaccaa tcccgtgaaa gaa 23
<210>25
<211>20
<212>DNA
<213>Homo sapiens
<400>25
gcgcttatcg catacggcta 20
<210>26
<211>18
<212>DNA
<213>Homo sapiens
<400>26
gaggtgcagg ccatgctc 18
<210>27
<211>21
<212>DNA
<213>Homo sapiens
<400>27
ggacgagacc atgaaggagt t 21
<210>28
<211>18
<212>DNA
<213>Homo sapiens
<400>28
gccatcaccc agcggatg 18
<210>29
<211>22
<212>DNA
<213>Homo sapiens
<400>29
gcacacacct gtgattgtgg tc 22
<210>30
<211>26
<212>DNA
<213>Homo sapiens
<400>30
caagctacac atgatcgagc tataca 26
<210>31
<211>25
<212>DNA
<213>Homo sapiens
<400>31
atccccaact atagaacctg gaagc 25
<210>32
<211>25
<212>DNA
<213>Homo sapiens
<400>32
ccccagaagt ggaaactgag atgat 25
<210>33
<211>28
<212>DNA
<213>Homo sapiens
<400>33
ttggtcttct tttctgtaca gagagagt 28
<210>34
<211>24
<212>DNA
<213>Homo sapiens
<400>34
agagtgggta aagggatagg catc 24
<210>35
<211>23
<212>DNA
<213>Homo sapiens
<400>35
ccatcctcca gaacttctcc atg 23
<210>36
<211>28
<212>DNA
<213>Homo sapiens
<400>36
aattacaacc agagcttggc atattgta 28
<210>37
<211>32
<212>DNA
<213>Homo sapiens
<400>37
tcagaataac taatgtttgg aagttgtttt gt 32
<210>38
<211>28
<212>DNA
<213>Homo sapiens
<400>38
ttgaatgaaa acatcaggat tgtaagca 28
<210>39
<211>18
<212>DNA
<213>Homo sapiens
<400>39
gcttggagtg caagctca 18
<210>40
<211>25
<212>DNA
<213>Homo sapiens
<400>40
gtgtttgtca ctctcacagt tacac 25
<210>41
<211>22
<212>DNA
<213>Homo sapiens
<400>41
ctttcttgcc tgggatctcc ct 22
<210>42
<211>24
<212>DNA
<213>Homo sapiens
<400>42
acaccaagaa tcgatggaca tcaa 24
<210>43
<211>23
<212>DNA
<213>Homo sapiens
<400>43
agagattgaa cgtgtgattg gca 23
<210>44
<211>22
<212>DNA
<213>Homo sapiens
<400>44
ctttcttgcc tgggatctcc ct 22
<210>45
<211>17
<212>DNA
<213>Homo sapiens
<400>45
ccagatgggc tcacgct 17
<210>46
<211>19
<212>DNA
<213>Homo sapiens
<400>46
gggacccgag ttggaacta 19
<210>47
<211>19
<212>DNA
<213>Homo sapiens
<400>47
tcctccctca cctggtcga 19
<210>48
<211>19
<212>DNA
<213>Homo sapiens
<400>48
tgtccaagag accgttggg 19
<210>49
<211>18
<212>DNA
<213>Homo sapiens
<400>49
gtccccaccg ctgcttgc 18
<210>50
<211>21
<212>DNA
<213>Homo sapiens
<400>50
gcctacctta gggatgcgga a 21
<210>51
<211>19
<212>DNA
<213>Homo sapiens
<400>51
ccagcctccc ctcattcct 19
<210>52
<211>19
<212>DNA
<213>Homo sapiens
<400>52
cagacatgcg tccccacaa 19
<210>53
<211>28
<212>DNA
<213>Homo sapiens
<400>53
aattacaacc agagcttggc atattgta 28
<210>54
<211>27
<212>DNA
<213>Homo sapiens
<400>54
gcaaccttag gttctagttc attaggg 27
<210>55
<211>22
<212>DNA
<213>Homo sapiens
<400>55
ctcaagcaac tcacctgacg at 22
<210>56
<211>32
<212>DNA
<213>Homo sapiens
<400>56
aaaaagagag aaagaaataa tagcccacat ac 32
<210>57
<211>26
<212>DNA
<213>Homo sapiens
<400>57
agtcctctca agtctaatag caactg 26
<210>58
<211>25
<212>DNA
<213>Homo sapiens
<400>58
catttttgct aaggcttcac ctctt 25
<210>59
<211>24
<212>DNA
<213>Homo sapiens
<400>59
atcactcact ttgtgaccat tccg 24
<210>60
<211>24
<212>DNA
<213>Homo sapiens
<400>60
ttcacaaagc ggaagaatgt gtca 24
<210>61
<211>28
<212>DNA
<213>Homo sapiens
<400>61
ataggttgtt taaaggaatc tgggtcat 28
<210>62
<211>23
<212>DNA
<213>Homo sapiens
<400>62
ttcacagata agggtacgcc tct 23
<210>63
<211>28
<212>DNA
<213>Homo sapiens
<400>63
gggataaaac agtttctctt aagtggtg 28
<210>64
<211>25
<212>DNA
<213>Homo sapiens
<400>64
catctttctg cttctgcctg atgta 25
<210>65
<211>26
<212>DNA
<213>Homo sapiens
<400>65
aaaatcttgc cttacaatgt gtggag 26
<210>66
<211>28
<212>DNA
<213>Homo sapiens
<400>66
cttccatctt acatggcacc catttata 28
<210>67
<211>22
<212>DNA
<213>Homo sapiens
<400>67
aacagtaagg gatccctctg gg 22
<210>68
<211>25
<212>DNA
<213>Homo sapiens
<400>68
gctgcaggaa agaatcattc tcaaa 25
<210>69
<211>19
<212>DNA
<213>Homo sapiens
<400>69
acaggtcagc gtgggaaga 19
<210>70
<211>27
<212>DNA
<213>Homo sapiens
<400>70
aataagtcag ctattgcagt agcctag 27
<210>71
<211>26
<212>DNA
<213>Homo sapiens
<400>71
gagtgcccat tgaactacac attaca 26
<210>72
<211>28
<212>DNA
<213>Homo sapiens
<400>72
atgttctttg aaaccctatg aacctgaa 28
<210>73
<211>31
<212>DNA
<213>Homo sapiens
<400>73
gttgatgctt ttgaagaacg acataaaagt t 31
<210>74
<211>28
<212>DNA
<213>Homo sapiens
<400>74
tctttctggt aggacaaata ttggcaaa 28
<210>75
<211>30
<212>DNA
<213>Homo sapiens
<400>75
cacccagcca attttgagta tttttaaaag 30
<210>76
<211>27
<212>DNA
<213>Homo sapiens
<400>76
tcccaggtag gttgaatact acatctg 27
<210>77
<211>29
<212>DNA
<213>Homo sapiens
<400>77
tcaaaatctt caattgttcg aggcttaag 29
<210>78
<211>26
<212>DNA
<213>Homo sapiens
<400>78
gtgaatcatg ccagtgctgt attttt 26
<210>79
<211>30
<212>DNA
<213>Homo sapiens
<400>79
ggtaagggta cagaagatac atgataggtc 30
<210>80
<211>24
<212>DNA
<213>Homo sapiens
<400>80
ttctgtcaag cagaaaatgc aagg 24
<210>81
<211>24
<212>DNA
<213>Homo sapiens
<400>81
atgaagccca ccaaacagta aacc 24
<210>82
<211>30
<212>DNA
<213>Homo sapiens
<400>82
attttcttca aaataacgtg agggtagaga 30
<210>83
<211>22
<212>DNA
<213>Homo sapiens
<400>83
tggtctgaaa accgattgtg gt 22
<210>84
<211>22
<212>DNA
<213>Homo sapiens
<400>84
gtgacctggc caagaagaag at 22
<210>85
<211>23
<212>DNA
<213>Homo sapiens
<400>85
gctcaagctg gaggacttct ttg 23
<210>86
<211>23
<212>DNA
<213>Homo sapiens
<400>86
tgcagatttg ccaacaggat ctt 23
<210>87
<211>22
<212>DNA
<213>Homo sapiens
<400>87
ctcaacaccc aaggagccca tt 22
<210>88
<211>24
<212>DNA
<213>Homo sapiens
<400>88
cgtctgaatg atgcagctct gatc 24
<210>89
<211>28
<212>DNA
<213>Homo sapiens
<400>89
tctgggctaa taggactact tctaatct 28
<210>90
<211>23
<212>DNA
<213>Homo sapiens
<400>90
caccaggtgg tggattctta agt 23
<210>91
<211>21
<212>DNA
<213>Homo sapiens
<400>91
caggctcagg gtcaatcaca g 21
<210>92
<211>17
<212>DNA
<213>Homo sapiens
<400>92
cgcggatggc gctgagg 17
<210>93
<211>17
<212>DNA
<213>Homo sapiens
<400>93
cacctcgccg cggtact 17
<210>94
<211>25
<212>DNA
<213>Homo sapiens
<400>94
ggtgatagtg ggttttcagt gaacg 25
<210>95
<211>30
<212>DNA
<213>Homo sapiens
<400>95
aaattggatg tgaaaaggaa gtttgctatc 30
<210>96
<211>23
<212>DNA
<213>Homo sapiens
<400>96
cgtgatggtg cacacctgta att 23
<210>97
<211>18
<212>DNA
<213>Homo sapiens
<400>97
gggaagccct ccccatcg 18
<210>98
<211>27
<212>DNA
<213>Homo sapiens
<400>98
ctggatacca gaaagactaa gctccat 27
<210>99
<211>28
<212>DNA
<213>Homo sapiens
<400>99
aatctcatag atgactgcct ctgtgtat 28
<210>100
<211>23
<212>DNA
<213>Homo sapiens
<400>100
ccaactgggc cttttatcct gac 23
<210>101
<211>19
<212>DNA
<213>Homo sapiens
<400>101
ggtctctcag aggcaggaa 19
<210>102
<211>30
<212>DNA
<213>Homo sapiens
<400>102
tccaaaatat cactttccat aaaagcaagg 30
<210>103
<211>25
<212>DNA
<213>Homo sapiens
<400>103
atcgtggcgc attatctctt acatc 25
<210>104
<211>28
<212>DNA
<213>Homo sapiens
<400>104
gtattcaaaa atgtacttca gggcttgg 28
<210>105
<211>23
<212>DNA
<213>Homo sapiens
<400>105
atttccaatc actgggagag gag 23
<210>106
<211>23
<212>DNA
<213>Homo sapiens
<400>106
gtcaaggtcc tttgggtcaa tca 23
<210>107
<211>21
<212>DNA
<213>Homo sapiens
<400>107
acccaccctt ggtttttctc a 21
<210>108
<211>28
<212>DNA
<213>Homo sapiens
<400>108
cattactcct tgacctgtta aacatccg 28
<210>109
<211>32
<212>DNA
<213>Homo sapiens
<400>109
cttgggaatg agatagtttc tgaatttaat gt 32
<210>110
<211>21
<212>DNA
<213>Homo sapiens
<400>110
acccaccctt ggtttttctc a 21
<210>111
<211>19
<212>DNA
<213>Homo sapiens
<400>111
gggctacccc gttctgtcc 19
<210>112
<211>20
<212>DNA
<213>Homo sapiens
<400>112
gctgcagcac ttcagcttct 20
<210>113
<211>19
<212>DNA
<213>Homo sapiens
<400>113
tgtccagagg agcccattt 19
<210>114
<211>17
<212>DNA
<213>Homo sapiens
<400>114
gcctgggcaa gaagtcg 17
<210>115
<211>20
<212>DNA
<213>Homo sapiens
<400>115
ggcttgacaa gaggccctga 20
<210>116
<211>17
<212>DNA
<213>Homo sapiens
<400>116
gtccaggccg tgtccaa 17
<210>117
<211>20
<212>DNA
<213>Homo sapiens
<400>117
cccagctgga tgagctgcta 20
<210>118
<211>20
<212>DNA
<213>Homo sapiens
<400>118
ccatctggga aacagtgcag 20
<210>119
<211>30
<212>DNA
<213>Homo sapiens
<400>119
tccaaaatat cactttccat aaaagcaagg 30
<210>120
<211>30
<212>DNA
<213>Homo sapiens
<400>120
acgaatgctc tactgtcatt tctaaccata 30
<210>121
<211>28
<212>DNA
<213>Homo sapiens
<400>121
taactcacca gccctctgat ctataaag 28
<210>122
<211>25
<212>DNA
<213>Homo sapiens
<400>122
gcctacagca tggatgtgat tactg 25
<210>123
<211>28
<212>DNA
<213>Homo sapiens
<400>123
gcatggactc agttgagagt taattcaa 28
<210>124
<211>28
<212>DNA
<213>Homo sapiens
<400>124
ccagcagtgt tctttccttc actttata 28
<210>125
<211>27
<212>DNA
<213>Homo sapiens
<400>125
ctgaaggact actacctctt ctacctg 27
<210>126
<211>24
<212>DNA
<213>Homo sapiens
<400>126
tctcctgact gtcatcccta ttgg 24
<210>127
<211>32
<212>DNA
<213>Homo sapiens
<400>127
tgtgtacatt acctaaatac aaagaagaat gt 32
<210>128
<211>28
<212>DNA
<213>Homo sapiens
<400>128
tctatttctg tttgcaggct atacagtt 28
<210>129
<211>28
<212>DNA
<213>Homo sapiens
<400>129
ccatttttct cttctctgag ctaacatg 28
<210>130
<211>27
<212>DNA
<213>Homo sapiens
<400>130
gtagatgtcc tcatgcatat cttgtgt 27
<210>131
<211>25
<212>DNA
<213>Homo sapiens
<400>131
cctggcttta aatcctcgaa cacaa 25
<210>132
<211>26
<212>DNA
<213>Homo sapiens
<400>132
ttggtgtcaa agtgtcactg aactaa 26
<210>133
<211>20
<212>DNA
<213>Homo sapiens
<400>133
ctgaaaaaca accattggcc 20
<210>134
<211>25
<212>DNA
<213>Homo sapiens
<400>134
tacgtcttca aggtgtaaaa tgctc 25
<210>135
<211>21
<212>DNA
<213>Homo sapiens
<400>135
ccttcccttt gtgacttgaa g 21
<210>136
<211>26
<212>DNA
<213>Homo sapiens
<400>136
gtatttatac tctagaaggg ggcaca 26
<210>137
<211>22
<212>DNA
<213>Homo sapiens
<400>137
cacatactgt ccaattcccc tg 22
<210>138
<211>28
<212>DNA
<213>Homo sapiens
<400>138
gtgtaaatgt atgattttat gcaggttt 28
<210>139
<211>28
<212>DNA
<213>Homo sapiens
<400>139
ggaattgact gtctttttga aaagttat 28
<210>140
<211>23
<212>DNA
<213>Homo sapiens
<400>140
ttgacatgat ttgggataga gga 23
<210>141
<211>30
<212>DNA
<213>Homo sapiens
<400>141
tattttaaca tgttactctt tcttgtttca 30
<210>142
<211>22
<212>DNA
<213>Homo sapiens
<400>142
tgctttgtgg atgttacaca gg 22
<210>143
<211>26
<212>DNA
<213>Homo sapiens
<400>143
tgatctccta gaagacagca aatacc 26
<210>144
<211>27
<212>DNA
<213>Homo sapiens
<400>144
atttttgatc aagttgtgag aagaaat 27
<210>145
<211>30
<212>DNA
<213>Homo sapiens
<400>145
ataggagatt caattataag gacaatacag 30
<210>146
<211>19
<212>DNA
<213>Homo sapiens
<400>146
cttctcctgc aggtgacca 19
<210>147
<211>30
<212>DNA
<213>Homo sapiens
<400>147
aaaaaatata cttatttacg cttgaacctc 30
<210>148
<211>18
<212>DNA
<213>Homo sapiens
<400>148
tgcccaaacc tggtgatg 18
<210>149
<211>26
<212>DNA
<213>Homo sapiens
<400>149
ttttgatcac attgtaagaa gaaacc 26
<210>150
<211>19
<212>DNA
<213>Homo sapiens
<400>150
gcccgaaaac accttcatc 19
<210>151
<211>18
<212>DNA
<213>Homo sapiens
<400>151
cgcctcaaca gccacatg 18
<210>152
<211>18
<212>DNA
<213>Homo sapiens
<400>152
acgcaggcga tgttgtcc 18
<210>153
<211>18
<212>DNA
<213>Homo sapiens
<400>153
aggccaccaa agggtacc 18
<210>154
<211>19
<212>DNA
<213>Homo sapiens
<400>154
agggacctgc agagctctg 19
<210>155
<211>18
<212>DNA
<213>Homo sapiens
<400>155
gcagatccct ggacaggc 18
<210>156
<211>23
<212>DNA
<213>Homo sapiens
<400>156
tcccaataaa agtgactctc agc 23
<210>157
<211>20
<212>DNA
<213>Homo sapiens
<400>157
gtgcaattca accctggttc 20
<210>158
<211>18
<212>DNA
<213>Homo sapiens
<400>158
gccgatgacc tgcagaag 18
<210>159
<211>18
<212>DNA
<213>Homo sapiens
<400>159
gcggacatgg aggacgtg 18
<210>160
<211>18
<212>DNA
<213>Homo sapiens
<400>160
atggtggatt tcgctggc 18
<210>161
<211>18
<212>DNA
<213>Homo sapiens
<400>161
agccatgatt gtggcaca 18
<210>162
<211>21
<212>DNA
<213>Homo sapiens
<400>162
ggcatgacaa ttgcttgaat c 21
<210>163
<211>19
<212>DNA
<213>Homo sapiens
<400>163
cctgggctag gtgtagggg 19
<210>164
<211>18
<212>DNA
<213>Homo sapiens
<400>164
agggtgagct ctgtgggc 18
<210>165
<211>26
<212>DNA
<213>Homo sapiens
<400>165
ctgtacagag agagtctaca gggaga 26
<210>166
<211>20
<212>DNA
<213>Homo sapiens
<400>166
gtggggaatg gatgaaattt 20
<210>167
<211>23
<212>DNA
<213>Homo sapiens
<400>167
aaataccccc aacataccag atc 23
<210>168
<211>26
<212>DNA
<213>Homo sapiens
<400>168
ttttcccact atcattgatt atttcc 26
<210>169
<211>26
<212>DNA
<213>Homo sapiens
<400>169
caaatttgtg tcttctgttc tcaaag 26
<210>170
<211>19
<212>DNA
<213>Homo sapiens
<400>170
ggattgtaag caccccctg 19
<210>171
<211>20
<212>DNA
<213>Homo sapiens
<400>171
aaggaccaca aaaggatcca 20
<210>172
<211>26
<212>DNA
<213>Homo sapiens
<400>172
ccctatgttt gttattttca ggaaaa 26
<210>173
<211>18
<212>DNA
<213>Homo sapiens
<400>173
tctccctcat gacgctgc 18
<210>174
<211>22
<212>DNA
<213>Homo sapiens
<400>174
ttcctgatca aaatggagaa gg 22
<210>175
<211>21
<212>DNA
<213>Homo sapiens
<400>175
gaacgtgtga ttggcagaaa c 21
<210>176
<211>20
<212>DNA
<213>Homo sapiens
<400>176
ggaagaggag cattgaggac 20
<210>177
<211>20
<212>DNA
<213>Homo sapiens
<400>177
caggtcagcc accactatgc 20
<210>178
<211>21
<212>DNA
<213>Homo sapiens
<400>178
gtgtctttgc tttcctggtg a 21
<210>179
<211>18
<212>DNA
<213>Homo sapiens
<400>179
gcagcaggtt gcccagcc 18
<210>180
<211>18
<212>DNA
<213>Homo sapiens
<400>180
gccttcgcca accactcc 18
<210>181
<211>18
<212>DNA
<213>Homo sapiens
<400>181
ccgaaaccca ggatctgg 18
<210>182
<211>20
<212>DNA
<213>Homo sapiens
<400>182
tccaacagga gatcgacgac 20
<210>183
<211>22
<212>DNA
<213>Homo sapiens
<400>183
ggatgagctg ctaactgagc ac 22
<210>184
<211>18
<212>DNA
<213>Homo sapiens
<400>184
gcccccgcct gtaccctt 18
<210>185
<211>24
<212>DNA
<213>Homo sapiens
<400>185
tcccactatc attgattatt tccc 24
<210>186
<211>27
<212>DNA
<213>Homo sapiens
<400>186
ctctttaaag agctcttttg tctttca 27
<210>187
<211>23
<212>DNA
<213>Homo sapiens
<400>187
tttgtagata tgggacccgt aca 23
<210>188
<211>28
<212>DNA
<213>Homo sapiens
<400>188
gatctaagaa accaaatttt aggaactt 28
<210>189
<211>18
<212>DNA
<213>Homo sapiens
<400>189
tccttccagg caccacct 18
<210>190
<211>24
<212>DNA
<213>Homo sapiens
<400>190
cagaaactgc aaaaggagat tgat 24
<210>191
<211>20
<212>DNA
<213>Homo sapiens
<400>191
ggaggagctg accagtgaag 20
<210>192
<211>19
<212>DNA
<213>Homo sapiens
<400>192
gagaaggtgt ctgcgggag 19
<210>193
<211>25
<212>DNA
<213>Homo sapiens
<400>193
tctgggtcat acatgtggat atatg 25
<210>194
<211>20
<212>DNA
<213>Homo sapiens
<400>194
gcagtcgaca atagggcaaa 20
<210>195
<211>22
<212>DNA
<213>Homo sapiens
<400>195
tttggacctt atctggaaca gc 22
<210>196
<211>20
<212>DNA
<213>Homo sapiens
<400>196
cactacatca tacggcagcc 20
<210>197
<211>23
<212>DNA
<213>Homo sapiens
<400>197
gaacacaaac tcatgcaact ctg 23
<210>198
<211>23
<212>DNA
<213>Homo sapiens
<400>198
taaaggctga ctttccagac aac 23
Claims (6)
1. A multiplex PCR detection kit for detecting human drug gene polymorphism, which is characterized in that: comprises a sixty-six PCR amplification primer pair which is designed according to the polymorphic sites of VKORC1, UGT1A1, HLA-A, HLA-B, TPMT, NAT1, NAT2, G6PD, F5, F2, IFNL3, APOE, COMT, CYP1A2, CYP2B6, CYP2C19, CYP2C9, CYP2D6, CYP3A5, MTFHR, SLCO1B1 and DPYD genes and reacts in the same reaction system, and a sixty-six single-base extension primer which reacts in the same reaction system; wherein,
the sixty-six PCR amplification primer pairs comprise first to sixty-six amplification primer pairs, wherein forward primers of the first to sixty-six amplification primer pairs are sequentially shown as SEQ D NO 01-066, and reverse primers are sequentially shown as SEQ ID NO 067-132; the sixty-six single-base extension primers comprise first to sixteenth single-base extension primers which are sequentially shown as SEQ ID NO 133-198;
the using method comprises the following steps:
(1) taking the genomic DNA to be detected as a template, and carrying out PCR amplification reaction in a PCR reaction system containing a RingCap-Taq enzyme and a RingCap buffer by using the sixty-six PCR amplification primer pair to obtain a PCR amplification reaction solution;
(2) carrying out SAP reaction on the PCR amplification reaction solution and SAP enzyme to obtain SAP reaction solution;
(3) performing single-base extension reaction on the SAP reaction solution by using the sixty-six single-base extension primer pair;
(4) and (4) desalting the material obtained in the step (3) by using resin, loading the material on a SpectrocHIP chip, and performing mass spectrometry detection to obtain the material.
2. The multiplex PCR detection kit of claim 1, wherein: the conditions of the PCR amplification reaction include: 3min at 95 ℃ for 1 cycle; 45 cycles of 95 ℃ for 20s, 60 ℃ for 30s, 72 ℃ for 20 s; 3min at 72 ℃ for 1 cycle.
3. The multiplex PCR detection kit of claim 2, wherein: the conditions under which the SAP reacts include: 40min at 37 ℃, 5min at 85 ℃ and infinity at 4 ℃.
4. The multiplex PCR detection kit of claim 3, wherein: the conditions of the single base extension reaction include:
the first stage consists of 40 cycles: each cycle comprises:
30s at 94 ℃, 1 cycle, 5s at 94 ℃, 5s at 52 ℃, 5s at 80 ℃, 5 cycles;
the second stage is 1 cycle in total: 3min at 72 ℃.
5. The multiplex PCR detection kit according to any one of claims 1 to 4 wherein: the RingCap-Taq enzyme consists of Taq enzyme, DNA ligase and DNA end modification enzyme in a ratio of 0.8-1.2: 0.8-1.2.
6. The multiplex PCR detection kit of claim 5, wherein: the RingCap-Taq enzyme consists of Taq enzyme, DNA ligase and DNA end modifying enzyme in the ratio of 1 to 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810660244.0A CN108823301A (en) | 2018-06-25 | 2018-06-25 | It is a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810660244.0A CN108823301A (en) | 2018-06-25 | 2018-06-25 | It is a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108823301A true CN108823301A (en) | 2018-11-16 |
Family
ID=64138469
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810660244.0A Pending CN108823301A (en) | 2018-06-25 | 2018-06-25 | It is a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108823301A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109536605A (en) * | 2019-01-31 | 2019-03-29 | 深圳市第二人民医院 | The PCR primer combination and application of statins adverse reaction genotype polymorphism |
| CN109825572A (en) * | 2019-03-13 | 2019-05-31 | 陈向东 | A kind of kit for detecting gene polymorphism related to the sensitivity of propofol and its detection method |
| CN111057755A (en) * | 2019-06-14 | 2020-04-24 | 陕西九州医学检验有限公司 | Cardiovascular and cerebrovascular disease genetic risk assessment detection panel and application thereof |
| CN112522376A (en) * | 2020-12-21 | 2021-03-19 | 济南和合医学检验有限公司 | Primer group, kit and method for detecting gene polymorphism |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110243925A1 (en) * | 2005-04-13 | 2011-10-06 | Oncotest Gmbh | Gene Expression Profiles Being Predictive for the Response of Tumors to Pharmaceutically Effective Compounds |
| CN106498036A (en) * | 2016-09-30 | 2017-03-15 | 厦门飞朔生物技术有限公司 | A kind of construction method in the Drug Discovery SNP variations library for high-flux sequence detection and its application |
| CN107937487A (en) * | 2017-12-29 | 2018-04-20 | 北京诺诗康瀛基因技术股份有限公司 | It is a kind of to be used for the amplification of HLA A gene PCRs, the method for Genotyping, primer sets and kit |
-
2018
- 2018-06-25 CN CN201810660244.0A patent/CN108823301A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110243925A1 (en) * | 2005-04-13 | 2011-10-06 | Oncotest Gmbh | Gene Expression Profiles Being Predictive for the Response of Tumors to Pharmaceutically Effective Compounds |
| CN106498036A (en) * | 2016-09-30 | 2017-03-15 | 厦门飞朔生物技术有限公司 | A kind of construction method in the Drug Discovery SNP variations library for high-flux sequence detection and its application |
| CN107937487A (en) * | 2017-12-29 | 2018-04-20 | 北京诺诗康瀛基因技术股份有限公司 | It is a kind of to be used for the amplification of HLA A gene PCRs, the method for Genotyping, primer sets and kit |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109536605A (en) * | 2019-01-31 | 2019-03-29 | 深圳市第二人民医院 | The PCR primer combination and application of statins adverse reaction genotype polymorphism |
| CN109825572A (en) * | 2019-03-13 | 2019-05-31 | 陈向东 | A kind of kit for detecting gene polymorphism related to the sensitivity of propofol and its detection method |
| CN111057755A (en) * | 2019-06-14 | 2020-04-24 | 陕西九州医学检验有限公司 | Cardiovascular and cerebrovascular disease genetic risk assessment detection panel and application thereof |
| CN111057755B (en) * | 2019-06-14 | 2022-07-26 | 陕西九州医学检验有限公司 | Cardiovascular and cerebrovascular disease genetic risk assessment detection panel and application thereof |
| CN112522376A (en) * | 2020-12-21 | 2021-03-19 | 济南和合医学检验有限公司 | Primer group, kit and method for detecting gene polymorphism |
| CN112522376B (en) * | 2020-12-21 | 2022-07-19 | 济南和合医学检验有限公司 | Primer group, kit and method for detecting gene polymorphism |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112080563A (en) | Kit for detecting accurate medication genes of chronic diseases | |
| MX2013008846A (en) | Method for discovering pharmacogenomic biomarkers. | |
| CN108823301A (en) | It is a kind of for detecting the multiple PCR detection kit of people's drug gene polymorphism | |
| CN104673915A (en) | Rapid detection kit for gene single-nucleotide polymorphism site and method for rapid detection kit | |
| CN111808944A (en) | Gene detection method for children personalized medicine | |
| CN110511989A (en) | A kind of high-flux sequence method and its application of hypertension therapeutic pharmaceutical relevant gene | |
| CN110541033A (en) | composition for detecting EGFR gene mutation and detection method | |
| CN112553325B (en) | Guiding method and kit for personalized medicine gene of sufentanil | |
| CN113584161A (en) | Detection kit for fentanyl metabolic marker, detection method and application thereof | |
| CN111793682A (en) | Detection kit for polymorphism of gene related to individualized medication and application thereof | |
| CN112501283B (en) | Gene-guided method and kit for personalized drug use of carbamazepine | |
| CN106947815B (en) | Method and special primer for detecting accurate medication of aspirin and nitroglycerin | |
| CN107868824A (en) | Single base extension method detects primer system, the methods and applications of UGT1A1*6 gene pleiomorphisms | |
| EP4106872A1 (en) | Compositions and methods for assessing the efficacy of inhibitors of neurotransmitter transporters | |
| CN112592972A (en) | Early screening method and kit for diffuse toxic goiter susceptibility genes | |
| CN112680517B (en) | Guiding method and kit for oxaliplatin personalized medicine genes | |
| CN112646869B (en) | Guidance method and kit for atorvastatin personalized medicine genes | |
| CN112695094B (en) | Gene-guided method and kit for personalized medication of epirubicin | |
| CN109825590A (en) | Method, kit and the primer of tumour driving gene mutation and microsatellite instability joint-detection | |
| CN111197077A (en) | Kit for detecting medication guidance gene of nifedipine serving as antihypertensive drug | |
| CN112831558B (en) | Early screening method and kit for Crohn disease susceptibility genes | |
| CN111197080A (en) | Detection product for distinguishing nifedipine individualized medication type | |
| US20240417801A1 (en) | Compositions and methods for assessing the efficacy of polynucleotide delivery and cancer therapy | |
| CN111197078A (en) | Method for identifying individualized medication of nifedipine by mass spectrometry by detecting products | |
| JP5015547B2 (en) | Method and kit for predicting side effects of docetaxel therapy |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181116 |
|
| RJ01 | Rejection of invention patent application after publication |