CN107099602A - It is a kind of at the same detect statins metabolic gene multisite mutation kit - Google Patents
It is a kind of at the same detect statins metabolic gene multisite mutation kit Download PDFInfo
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- CN107099602A CN107099602A CN201710392093.0A CN201710392093A CN107099602A CN 107099602 A CN107099602 A CN 107099602A CN 201710392093 A CN201710392093 A CN 201710392093A CN 107099602 A CN107099602 A CN 107099602A
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- 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
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- C—CHEMISTRY; METALLURGY
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- 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
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- 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
- C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
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- 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
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/16—Primer sets for multiplex assays
Abstract
The present invention relates to a kind of while the kit of detection statins metabolic gene multisite mutation, including detection include APOE, SLCO1B1, CETP, ABCB1, the primer pair in mthfr gene site and probe pair.Present invention application MGB (minor groove binder) probes and real-time fluorescence PCR technology, have the advantages that high time saving convenience, sensitivity, Sample Positive coincidence rate and negative match-rate equal more than 99.5%；It is mainly used in the personalized medicine auxiliary diagnosis of statins：Simvastatin, Atorvastatin, Pravastatin etc..
Description
Technical field
It is more particularly to a kind of to detect that statins metabolic gene is more simultaneously the invention belongs to statins detection field
The kit of site mutation.
Background technology
Statins is to use extensive lipid-lowering medicine, cardiovascular and cerebrovascular disease can be effectively prevented and treated, in the U.S. more than 25%
Age be more than the people of 45 years old and take statins.Especially in the percutaneous Stent of coronary heart disease (PCI) afterwards secondary prevention,
Significant is prevented and treated to coronary heart disease using positive statin Lipid modulating.
Statins is with the rise of dosage, reduction LDL effect enhancing, but adverse reaction rate is also high simultaneously.
One main toxicity of Statins is myopathy, and myopathy caused by statin can occur for the patient for having 5~29%
(statininducedmyopathy, SIM), the appearance for be mainly shown as tired, powerless, myalgia, muscle cramp, having seriously is endangered
The rhabdomyolysis (rhabdomyolysis) of evil life, causes death.Myopathy influences compliance, and 33% flesh occurs
The patient of disease can be voluntarily discontinued, and cause dead, the cardiovascular event such as heart infarction again generation, and the existence to patient is constituted seriously
Threaten.Drug safety can be ensured according to gene diagnosis progress personalized medicine, medicine spending is saved, and be increasingly becoming clinic and controlled
The new trend for the treatment of.Especially detect that following 5 kinds representative genes have important medication guide meaning：
(1) ApoE belongs to apolipoprotein family, is mainly expressed in liver and brain tissue, and body is participated in by number of ways
Lipid-metabolism, is to influence the important internal factor of blood lipid level.The ApoE assignments of genes gene mapping mainly have two on No. 19 chromosomes of the mankind
SNP is planted, 3 kinds of haplotypes, respectively ApoE2 (388T-526T), ApoE3 (388T-526C) and ApoE4 are formed
(388C-526C).ApoE gene pleiomorphisms are considered as hyperlipoprotememia and the susceptible time of atherosclerotic vascular disease
Select gene.There are 6 kinds of different ApoE phenotypes in crowd：Three kinds of homozygotes (E2/2, E3/3, E4/4) and three kinds of heterozygotes (E3/2,
E4/2、E4/3).Wherein, ApoE3/3 is wild type (Wild-type).The statins lipid-lowering effect of ApoE2 carrier is good,
The lipid-lowering effect of ApoE4 carrier's statinses is poor.
(2) SLCO1B1 genes have very high genetic polymorphism.Saltant type SLCO1B1 genes cause the transhipment egg of coding
White vigor weakens, and shows as liver intake medicine ability reduction, causes statins blood concentration to rise, increase striated muscle is molten
Solve the occurrence risk of disease or myopathy.SLCO1B1*1a, * 1b, * 15 are common gene mutation types, and wherein * 1b, * 15 are statins
The Independent Decisiveness factor of class medicine main adverse reaction " rhabdomyolysis ".Mutator carrier sends out compared to unmutated person
About 20 times of the risk increase of myogenic toxicity.Take the mutation is detected before statins to occur muscle poison prediction and
Prevention is significant.
(3) CETP CETPs, the SNP (G/A) in the sites of genome First Intron Taq1B the 277th is polymorphic
Property has obvious influence to the reaction effect of statins.Wherein B1B1 genotype and B1B2 genotype individuals statinses
Regulating lipid is good, and B2B2 genotype individuals react weaker to treatment.
(4) ABCB1 Multidrug resistances 1, expressing protein is to have two site SNP polymorphic in drug transporters, gene order
Property statins metabolism is had a certain impact, the respectively site of genome 2677 and 3435 sites, the two sites are all
Saltant type individual is small using lipid-lowering effect during Atorvastatin.
(5) MTHFR hyperpietics, TT genotype, are treated with Pravastatin, lethal coronary heart disease and non-lethality occur
The risk of miocardial infarction is higher than CC genotype.CC genotype is the genotype of protectiveness, and risk is low.CT genotype, between the two
Between.
At present, it is domestic to have examination related to SLCO1B1 genetic polymorphism detections for statins metabolic gene APOE
Agent box, but for the detection kits of 5 kinds of genes (APOE, SLCO1B1, CETP, ABCBI, MTHFR), there is presently no any factory
Family releases.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of while detecting that many sites of statins metabolic gene are dashed forward
The kit of change, the kit application MGB probes and real-time fluorescence PCR technology, with time saving convenience, sensitivity height, sample sun
Property coincidence rate and negative match-rate it is equal more than 99.5% the advantages of；The personalized medicine auxiliary for being mainly used in statins is examined
It is disconnected：Simvastatin, Atorvastatin, Pravastatin etc..
The present invention's is a kind of while detecting the kit of statins metabolic gene multisite mutation, the kit bag
The primer pair and probe pair for detecting APOE, SLCO1B1, CETP, ABCBI, MTHFR site are included, particular sequence is as follows：
The kit also includes archaeal dna polymerase, UNG enzymes, dNTPs, magnesium ion and PCR buffer solutions.
The kit also includes positive reference substance and negative controls.
The concentration of the kit primer pair is 400nM；The concentration of probe pair is 300nM.
The kit be used for quantitative PCR reaction system be：Cumulative volume 25 μ l, 20 μ l PCR MIX and 5 μ l DNA moulds
Plate.
The kit be used for quantitative PCR reaction condition be：50℃2min；95℃10min；95 DEG C of 15s, 60 DEG C of 45s,
40 circulations.
Beneficial effect
Present invention application MGB probes and real-time fluorescence PCR technology, meet with time saving convenience, sensitivity height, Sample Positive
Rate and negative match-rate it is equal more than 99.5% the advantages of；It is mainly used in the personalized medicine auxiliary diagnosis of statins：It is pungent to cut down
Statin, Atorvastatin, Pravastatin etc..
Brief description of the drawings
Fig. 1 is the SLCO1B1 gene rs2306283 site primer positive reference substance result figures of embodiment 1；
Fig. 2 is the SLCO1B1 gene rs2306283 site primer negative controls result figures of embodiment 1；
Fig. 3 is the SLCO1B1 gene rs2306283 site primer sample to be tested result figures of embodiment 1；
Fig. 4 is the SLCO1B1 gene rs4149056 site primer positive reference substance result figures of embodiment 1；
Fig. 5 is the SLCO1B1 gene rs4149056 site primer negative controls result figures of embodiment 1；
Fig. 6 is the SLCO1B1 gene rs4149056 site primer sample to be tested result figures of embodiment 1；
Fig. 7 is the APOE gene rs429358 site primer positive reference substance result figures of embodiment 1；
Fig. 8 is the APOE gene rs429358 site primer negative controls result figures of embodiment 1；
Fig. 9 is the APOE gene rs429358 site primer sample to be tested result figures of embodiment 1；
Figure 10 is the APOE gene rs7412 site primer positive reference substance result figures of embodiment 1；
Figure 11 is the APOE gene rs7412 site primer negative controls result figures of embodiment 1；
Figure 12 is the APOE gene rs7412 site primer sample to be tested result figures of embodiment 1；
Figure 13 is the CETP gene Taq1B site primer positive reference substance result figures of embodiment 1；
Figure 14 is the CETP gene Taq1B site primer negative controls result figures of embodiment 1；
Figure 15 is the CETP gene Taq1B site primer sample to be tested result figures of embodiment 1；
Figure 16 is the ABCB1 gene rs1128503 site primer positive reference substance result figures of embodiment 1；
Figure 17 is the ABCB1 gene rs1128503 site primer negative controls result figures of embodiment 1；
Figure 18 is the ABCB1 gene rs1128503 site primer sample to be tested result figures of embodiment 1；
Figure 19 is the ABCB1 gene rs1045642 site primer positive reference substance result figures of embodiment 1；
Figure 20 is the ABCB1 gene rs1045642 site primer negative controls result figures of embodiment 1；
Figure 21 is the ABCB1 gene rs1045642 site primer sample to be tested result figures of embodiment 1；
Figure 22 is the mthfr gene rs1801131 site primer positive reference substance result figures of embodiment 1；
Figure 23 is the mthfr gene rs1801131 site primer negative controls result figures of embodiment 1；
Figure 24 is the mthfr gene rs1801131 site primer sample to be tested result figures of embodiment 1.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
The detection method and flow, rather than limitation the scope of the present invention.In addition, it is to be understood that reading in of the invention lecture
After appearance, those skilled in the art can make various changes or modifications to the present invention, and these equivalent form of values equally fall within the application
Appended claims limited range.
Embodiment 1
1st, nucleic acid DNA is extracted
1.1 sequentially add 200ul whole blood samples to be measured, 20ul Proteinase Ks, 400ul lysates in 1.5ml centrifuge tubes,
Whirlpool concussion is mixed after 56 DEG C of water-baths or metal bath 10min.
1.2 add 400ul isopropanols, and whirlpool, which shakes, adds 20ul magnetic beads after mixing in 10 seconds, whirlpool shakes 10 seconds, and room temperature is quiet
Put 10min.
1.3 are put into centrifuge tube on magnetic separation rack, stand 10s, be all adsorbed to magnetic bead on centrifugation tube wall (if from
There is liquid then to keep centrifuge tube integrally to be overturned 3-5 times on magnetic separation rack in heart lid, the magnetic bead in lid is also adsorbed
Onto tube wall), whole liquid are carefully sopped up, are careful not to encounter magnetic bead.
1.4 remove centrifuge tube, add 750ul cleaning solutions I, and be vortexed several seconds washing magnetic bead, then centrifuge tube is put into magnetic point
From on frame, 10s is stood, (centrifuge tube is kept if centrifuge tube has covered liquid on magnetic separation rack to magnetic bead is all adsorbed
It is overall reverse 3-5 times, the magnetic bead in lid is also adsorbed on tube wall), whole liquid are carefully sopped up, are careful not to encounter magnetic
Pearl (exhausts centrifuge tube bottom liquid) as far as possible.
1.5 repeat steps 1.4 are once.
1.6 remove centrifuge tube, add 750ul cleaning solutions II, and be vortexed several seconds washing magnetic bead, and centrifuge tube is put into Magnetic Isolation
On frame, 10s is stood, is all adsorbed (keeping centrifuge tube whole on magnetic separation rack if centrifuge tube has covered liquid to magnetic bead
Body is reverse 3-5 times, the magnetic bead in lid is also adsorbed on tube wall), whole liquid are carefully sopped up, are careful not to encounter magnetic bead
(exhausting centrifuge tube bottom liquid as far as possible).
1.7 repeat steps 1.6 are once.
1.8 keep centrifuge tube on magnetic separation rack, and room temperature places 10-15min, dries residual ethanol in centrifuge tube.
Note：The purpose of this step is to remove rinsing liquid remaining in adsorption column, and the residual of ethanol can shadow in rinsing liquid
Ring follow-up enzyme reaction (digestion, PCR etc.) experiment.
1.9 remove centrifuge tube, and 50ul eluents are added into centrifuge tube, and the concussion that is vortexed mixes magnetic bead, and in 56 DEG C of water-baths
Or 10min elution nucleic acid is placed in metal bath.
1.10 are put into centrifuge tube on magnetic separation rack, stand 10s, and all adsorbed to magnetic bead, solution becomes clarification.Carefully
Liquid in pipe is transferred completely into another centrifuge tube by ground, is careful not to be drawn onto magnetic bead.4 DEG C of gained nucleic acid solution is temporary standby
With if long-term preserve please be placed under the conditions of -20 DEG C.
2nd, reagent prepares
2.1 taking out kit from refrigerator, balance to room temperature, each component fully dissolves.
2.2 according to sample to be tested, negative control, positive control quantity, (reaction solution 19ul/ person-portion+1ul/ people in proportion
Part enzyme mixation) reaction solution and enzyme mixation of respective amount are taken, fully it is mixed into standby after PCR-MIX, brief centrifugation.
3rd, PCR is loaded
The PCR-MIX for mixing standby is dispensed into PCR reaction tubes by 20ul/ person-portions, treating for 5ul/ person-portions is sequentially added
Sample DNA, positive reference substance, negative controls are surveyed, covers and PCR amplification regions is transferred to after lid, brief centrifugation.
4th, PCR is expanded
PCR reaction tubes are put into amplification instrument sample cell by 4.1, and sample names to be checked, positive control, the moon are sequentially set by correspondence
Property control.
4.2 detection fluorescence channel selections
By taking the real-time fluorescence PCR instrument of ABI 7500 as an example：1) FAM Air conduct measurement wild types, 2 are selected) selection VIC Air conduct measurements
Saltant type, 3) selection ROX Air conduct measurement internal standards, 4) reference fluorescent selection none.It is 25ul to set Sample volume.
4.3 loop parameters are set
By taking the real-time fluorescence PCR instrument of ABI 7500 as an example：
Setting completed, preserves file, runs response procedures.
5th, interpretation of result
The genotype that sample statins to be checked is metabolized each gene is analyzed according to experimental result picture (Fig. 1-2 4), can be obtained such as
Lower result：
SEQUENCE LISTING
<110>Ningbo Mei Jing medical technology Co., Ltd
<120>It is a kind of at the same detect statins metabolic gene multisite mutation kit
<130> 1
<160> 35
<170> PatentIn version 3.3
<210> 1
<211> 22
<212> DNA
<213>Artificial sequence
<400> 1
ttcttacagt tacaggtatt ct 22
<210> 2
<211> 19
<212> DNA
<213>Artificial sequence
<400> 2
tatctcaggt gatgctcta 19
<210> 3
<211> 22
<212> DNA
<213>Artificial sequence
<400> 3
aactaatatc aattcatcag aa 22
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<211> 22
<212> DNA
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aactaatatc gattcatcag aa 22
<210> 5
<211> 22
<212> DNA
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<400> 5
ataggttgtt taaaggaatc tg 22
<210> 6
<211> 22
<212> DNA
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<400> 6
ttagcgaaat catcaatgta ag 22
<210> 7
<211> 20
<212> DNA
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<400> 7
tggatatatg tgttcatggg 20
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<211> 20
<212> DNA
<213>Artificial sequence
<400> 8
tggatatatg cgttcatggg 20
<210> 9
<211> 17
<212> DNA
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<400> 9
tgtccaagga gctgcag 17
<210> 10
<211> 16
<212> DNA
<213>Artificial sequence
<400> 10
agctcctcgg tgctct 16
<210> 11
<211> 14
<212> DNA
<213>Artificial sequence
<400> 11
aggacgtgtg cggc 14
<210> 12
<211> 14
<212> DNA
<213>Artificial sequence
<400> 12
aggacgtgcg cggc 14
<210> 13
<211> 16
<212> DNA
<213>Artificial sequence
<400> 13
caagctgcgt aagcgg 16
<210> 14
<211> 15
<212> DNA
<213>Artificial sequence
<400> 14
gcggatggcg ctgag 15
<210> 15
<211> 14
<212> DNA
<213>Artificial sequence
<400> 15
tgcagaagcg cctg 14
<210> 16
<211> 14
<212> DNA
<213>Artificial sequence
<400> 16
tgcagaagtg cctg 14
<210> 17
<211> 23
<212> DNA
<213>Artificial sequence
<400> 17
tagggatttg tgtttgtctg cga 23
<210> 18
<211> 21
<212> DNA
<213>Artificial sequence
<400> 18
tgactttggc catagagtga c 21
<210> 19
<211> 18
<212> DNA
<213>Artificial sequence
<400> 19
ttcgagttag ggttcaga 18
<210> 20
<211> 18
<212> DNA
<213>Artificial sequence
<400> 20
ttcaagttag ggttcaga 18
<210> 21
<211> 21
<212> DNA
<213>Artificial sequence
<400> 21
tctgtgaatt gccttgaagt t 21
<210> 22
<211> 20
<212> DNA
<213>Artificial sequence
<400> 22
atcagctgga ctgttgtgct 20
<210> 23
<211> 17
<212> DNA
<213>Artificial sequence
<400> 23
ttgaagggtc tgaacct 17
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<211> 17
<212> DNA
<213>Artificial sequence
<400> 24
ttgaagggcc tgaacct 17
<210> 25
<211> 22
<212> DNA
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ttgcctatgg agacaacagc cg 22
<210> 26
<211> 22
<212> DNA
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<400> 26
ttgaagagag acttacatta gg 22
<210> 27
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<212> DNA
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<400> 27
aagagatcgt gagggcag 18
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<211> 18
<212> DNA
<213>Artificial sequence
<400> 28
aagagattgt gagggcag 18
<210> 29
<211> 20
<212> DNA
<213>Artificial sequence
<400> 29
ttctacctga agagcaagtc 20
<210> 30
<211> 19
<212> DNA
<213>Artificial sequence
<400> 30
tttgtgacca ttccggttt 19
<210> 31
<211> 19
<212> DNA
<213>Artificial sequence
<400> 31
ccagtgaaga aagtgtctt 19
<210> 32
<211> 19
<212> DNA
<213>Artificial sequence
<400> 32
ccagtgaagc aagtgtctt 19
<210> 33
<211> 19
<212> DNA
<213>Artificial sequence
<400> 33
tacagcttca ccaccacgg 19
<210> 34
<211> 21
<212> DNA
<213>Artificial sequence
<400> 34
gtcaggcagc tcgtagctct t 21
<210> 35
<211> 27
<212> DNA
<213>Artificial sequence
<400> 35
aaggagaagc tgtgctacgt tgacatt 27
Claims (6)
1. it is a kind of while detecting the kit of statins metabolic gene multisite mutation, it is characterised in that：The kit
Include APOE, SLCO1B1, CETP, ABCB1, the primer pair in mthfr gene site and probe pair including detection, particular sequence is such as
Under：
SLCO1B1 rs2306283 sites：
Q201-F1：TTCTTACAGTTACAGGTATTCT；
Q201-R1：TATCTCAGGTGATGCTCTA；
Q201-WP1：AACTAATATCAATTCATCAGAA；
Q201-MP1：AACTAATATCGATTCATCAGAA；
SLCO1B1 rs4149056 sites：
Q202-F1：ATAGGTTGTTTAAAGGAATCTG；
Q202-R1：TTAGCGAAATCATCAATGTAAG；
Q202-WP1：TGGATATATGTGTTCATGGG；
Q202-MP1：TGGATATATGCGTTCATGGG；
APOE rs429358 sites：
Q301-F1：TGTCCAAGGAGCTGCAG；
Q301-R1：AGCTCCTCGGTGCTCT；
Q301-WP1：AGGACGTGTGCGGC；
Q301-MP1：AGGACGTGCGCGGC；
APOE rs7412 sites：
Q302-F1：CAAGCTGCGTAAGCGG；
Q302-R1：GCGGATGGCGCTGAG；
Q302-WP1：TGCAGAAGCGCCTG；
Q302-MP1：TGCAGAAGTGCCTG；
CETP Taq1B sites：
Q401-F1：TAGGGATTTGTGTTTGTCTGCGA；
Q401-R1：TGACTTTGGCCATAGAGTGAC；
Q401-WP1：TTCGAGTTAGGGTTCAGA；
Q401-MP1：TTCAAGTTAGGGTTCAGA；
ABCB1 rs1128503 sites：
Q501-F1：TCTGTGAATTGCCTTGAAGTT；
Q501-R1：ATCAGCTGGACTGTTGTGCT；
Q501-WP1：TTGAAGGGTCTGAACCT；
Q501-MP1：TTGAAGGGCCTGAACCT；
ABCB1 rs1045642 sites：
Q502-F1：TTGCCTATGGAGACAACAGCCG；
Q502-R1：TTGAAGAGAGACTTACATTAGG；
Q502-WP1：AAGAGATCGTGAGGGCAG；
Q502-MP1：AAGAGATTGTGAGGGCAG；
MTHFR rs1801131 sites：
Q601-F1：TTCTACCTGAAGAGCAAGTC；
Q601-R1：TTTGTGACCATTCCGGTTT；
Q601-WP1：CCAGTGAAGAAAGTGTCTT；
Q601-MP1：CCAGTGAAGCAAGTGTCTT.
2. it is according to claim 1 a kind of at the same detect statins metabolic gene multisite mutation kit, its
It is characterised by：The kit also includes interior label primer pair and probe, and particular sequence is as follows：
Q701-F1：TACAGCTTCACCACCACGG；
Q701-R1：GTCAGGCAGCTCGTAGCTCTT；
Q701-P：AAGGAGAAGCTGTGCTACGTTGACATT.
3. it is according to claim 1 a kind of at the same detect statins metabolic gene multisite mutation kit, its
It is characterised by：The kit also includes archaeal dna polymerase, UNG enzymes, dNTPs, magnesium ion and PCR buffer solutions.
4. it is according to claim 1 a kind of at the same detect statins metabolic gene multisite mutation kit, its
It is characterised by：The concentration of the kit primer pair is 400nM；The concentration of probe pair is 300nM.
5. it is according to claim 1 a kind of at the same detect statins metabolic gene multisite mutation kit, its
It is characterised by：The kit be used for quantitative PCR reaction system be：Cumulative volume 25 μ l, 20 μ l PCR MIX and 5 μ l DNA moulds
Plate.
6. it is according to claim 1 a kind of at the same detect statins metabolic gene multisite mutation kit, its
It is characterised by：The kit be used for quantitative PCR reaction condition be：50℃2min；95℃10min；95 DEG C of 15s, 60 DEG C
45s, 40 circulations.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710392093.0A CN107099602A (en) | 2017-05-27 | 2017-05-27 | It is a kind of at the same detect statins metabolic gene multisite mutation kit |
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| CN201710392093.0A CN107099602A (en) | 2017-05-27 | 2017-05-27 | It is a kind of at the same detect statins metabolic gene multisite mutation kit |
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| Publication Number | Publication Date |
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| CN107099602A true CN107099602A (en) | 2017-08-29 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710392093.0A Pending CN107099602A (en) | 2017-05-27 | 2017-05-27 | It is a kind of at the same detect statins metabolic gene multisite mutation kit |
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN110205370A (en) * | 2019-05-14 | 2019-09-06 | 南京派森诺基因科技有限公司 | Primer combination, kit and method for instructing Pravastatin drug personalized medicine related gene to detect |
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| CN108546747A (en) * | 2017-12-19 | 2018-09-18 | 美因健康科技（北京）有限公司 | The method and primer and probe used of Taqman probe in detecting APOE |
| CN108004315A (en) * | 2017-12-22 | 2018-05-08 | 佰世凯（杭州）生物科技有限公司 | Appraisal procedure for Alzheimer's disease risk |
| CN108977523A (en) * | 2018-04-28 | 2018-12-11 | 东莞博盛生物科技有限公司 | For detecting primer pair, probe and the kit of SLCO1B1 521T > C gene pleiomorphism |
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| CN110305955A (en) * | 2019-05-05 | 2019-10-08 | 上海派森诺生物科技股份有限公司 | Primer combination of probe, kit and application for instructing Simvastatin drug personalized medicine related gene to detect |
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| CN110241197A (en) * | 2019-05-28 | 2019-09-17 | 南京派森诺基因科技有限公司 | Primer combination of probe and kit and application for instructing Atorvastatin drug personalized medicine related gene to detect |
| CN110205372A (en) * | 2019-05-31 | 2019-09-06 | 大连美纳医学检验实验室有限公司 | Simvastatin, Atorvastatin pharmaceutical relevant gene detection primer group, kit and detection method |
| CN110157797A (en) * | 2019-06-05 | 2019-08-23 | 北京益序医疗科技有限公司 | A kind of probe groups and kit for statins individuation genetic test |
| CN110387412A (en) * | 2019-07-08 | 2019-10-29 | 上海派森诺生物科技股份有限公司 | Primer combination and kit and method for instructing Rosuvastatin drug personalized medicine related gene to detect |
| CN113528629A (en) * | 2021-06-15 | 2021-10-22 | 湖南菲思特精准医疗科技有限公司 | Detection kit for methotrexate metabolic marker, detection method and application thereof |
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