CN108949532A - For detecting the kit of COX-2 gene pleiomorphism - Google Patents
For detecting the kit of COX-2 gene pleiomorphism Download PDFInfo
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- CN108949532A CN108949532A CN201811191253.6A CN201811191253A CN108949532A CN 108949532 A CN108949532 A CN 108949532A CN 201811191253 A CN201811191253 A CN 201811191253A CN 108949532 A CN108949532 A CN 108949532A
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- 101150071146 COX2 gene Proteins 0.000 title claims abstract description 33
- 238000001514 detection method Methods 0.000 claims abstract description 24
- 239000000463 material Substances 0.000 claims abstract description 15
- 230000008014 freezing Effects 0.000 claims abstract description 14
- 238000007710 freezing Methods 0.000 claims abstract description 14
- 239000006260 foam Substances 0.000 claims abstract description 13
- 239000003153 chemical reaction reagent Substances 0.000 claims description 40
- 238000006243 chemical reaction Methods 0.000 claims description 26
- 239000013641 positive control Substances 0.000 claims description 19
- 239000000523 sample Substances 0.000 claims description 15
- 239000002773 nucleotide Substances 0.000 claims description 13
- 125000003729 nucleotide group Chemical group 0.000 claims description 13
- 102000054765 polymorphisms of proteins Human genes 0.000 claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 9
- 239000013642 negative control Substances 0.000 claims description 8
- 239000011148 porous material Substances 0.000 claims description 8
- 238000007789 sealing Methods 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 239000013612 plasmid Substances 0.000 claims description 6
- 238000011144 upstream manufacturing Methods 0.000 claims description 6
- 102000006943 Uracil-DNA Glycosidase Human genes 0.000 claims description 5
- 108010072685 Uracil-DNA Glycosidase Proteins 0.000 claims description 5
- 108020005196 Mitochondrial DNA Proteins 0.000 claims description 4
- 238000009432 framing Methods 0.000 claims description 4
- 101150040913 DUT gene Proteins 0.000 claims description 3
- 108010006785 Taq Polymerase Proteins 0.000 claims description 3
- 238000012986 modification Methods 0.000 claims description 3
- 230000004048 modification Effects 0.000 claims description 3
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
- 230000003000 nontoxic effect Effects 0.000 abstract description 3
- 238000004140 cleaning Methods 0.000 abstract description 2
- 230000006835 compression Effects 0.000 abstract description 2
- 238000007906 compression Methods 0.000 abstract description 2
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 abstract description 2
- 238000012546 transfer Methods 0.000 abstract description 2
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 description 8
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 description 8
- 238000010586 diagram Methods 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 102100038277 Prostaglandin G/H synthase 1 Human genes 0.000 description 2
- 108050003243 Prostaglandin G/H synthase 1 Proteins 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- 230000003416 augmentation Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000006555 catalytic reaction Methods 0.000 description 2
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- 238000005119 centrifugation Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 108010017796 epoxidase Proteins 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 229940094443 oxytocics prostaglandins Drugs 0.000 description 2
- 238000012257 pre-denaturation Methods 0.000 description 2
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- 150000003180 prostaglandins Chemical class 0.000 description 2
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
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- 102100037611 Lysophospholipase Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010058864 Phospholipases A2 Proteins 0.000 description 1
- 229960001138 acetylsalicylic acid Drugs 0.000 description 1
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- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
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- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000012631 diagnostic technique Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
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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
- C12Q1/6876—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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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/136—Screening for pharmacological compounds
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
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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
- C12Q2600/158—Expression markers
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Abstract
The invention discloses the kits for detecting COX-2 gene pleiomorphism, kit includes box body, box cover, foam mat, freezing box rack, frost box, box cover, by being hinged and connected, is equipped with the first groove with box body in box body, freezing box rack is vertically arranged at the middle position of the first groove, foam mat is located in box body, kit of the present invention is suitable for clinical a variety of sample type detections, has high specificity, high sensitivity, the advantages that experimental period is short, at low cost；The thorny problem of kit during transportation is not only solved, and during the test, can quickly be taken, it is easy to operate；Kit body uses 3-tier architecture, internal layer is PS material, safe and non-toxic, high temperature resistant, easy cleaning, insulating layer are PU material, have the function of cryogenic heat transfer, temperature and humidity can be kept to balance for a long time, and it is shockproof compression-resistant, waterproof and dampproof, outer layer is PE material, and chemical stability is strong, safety and environmental protection, wearability are good.
Description
Technical field
The present invention relates to vitro diagnostic techniques fields, and in particular to for detecting the kit of COX-2 gene pleiomorphism.
Background technique
Epoxidase is the key enzyme for synthesizing various prostaglandins (PGs), and the phosphatide in cell membrane is under phospholipase A2 catalysis
Arachidonic acid is released, the latter forms PGH2 under the catalysis of epoxidase, and PGH2 is again respectively in corresponding prostaglandin isomerase
Lower synthesis PGE, PGF, PGD, PGI of effect etc., there are two types of isodynamic enzyme COX-1 and COX-2 by COX, and the human body COX-2 assignment of genes gene mapping is in dye
The area 1q25.2~q25.3 of colour solid, length about 8.3kb include 10 exons and 9 intrones, encode 604 amino altogether
Acid.COX-2 is Inducing enzyme, under physiological conditions, be can't detect in most of tissues and cell, under ex vivo, many thorns
Swash object, such as growth factor, the agent of inflammatory cytokine tumor inducing can stimulate the COX-2 of cell to express, COX-2 is athero- in artery
It is expressed in patch, aspirin dose needs to reach 170 times of inhibition COX-1 dosage, could block COX-2, low dose of Ah Si
The TXA2 that woods cannot completely inhibit COX-2 induction is generated and one of the reason of aspirin resistance, COX-2 gene promoter
Sub- G-765C polymorphic site is research shows that GC heterozygous genes carrier is compared with GG homozygous gene carrier, the former is to Ah Si
The insensitive probability of woods drug effect is 3.872 times of the latter.
Therefore, COX-2 gene polymorphic sexually revises the combination that may potentially influence certain factor, and the transcription for changing gene is living
Property, and then the expression of Cycloxygenase is influenced, it can be individual to the susceptible of certain diseases by the detection to COX-2 gene pleiomorphism
Property diagnosis help is provided.
Summary of the invention
The purpose of the present invention is to provide the kits for detecting COX-2 gene pleiomorphism, solve currently on the market
For COX-2 genetic polymorphism detection not strong and complicated for operation problem at high cost, time-consuming, specific, kit is small in size, behaviour
Make simply, to be readily transported and carry and use.
To achieve the above object, the invention provides the following technical scheme: being used to detect the reagent of COX-2 gene pleiomorphism
Box, kit include box body, box cover, foam mat, freezing box rack, frost box, box cover and box body by being hinged and connected, box cover and
Box body is closed by fastener lid, and the first groove is equipped in box body, and freezing box rack is vertically arranged at the middle position of the first groove, sponge
Liner is located in box body, and foam mat is equipped with multiple reagent pores and multiple Reagent Tubes equipped with detection reagent, reagent are housed
Pipe includes Reagent Tube, the Reagent Tube equipped with saltant type positive control, the examination equipped with negative control equipped with wild-type positive control
Agent pipe, the Reagent Tube equipped with G-765C single nucleotide polymorphisms detection reaction solution；Reagent pore is annular on the foam mat
Arrangement, above-mentioned examines equipped with wild-type positive control, saltant type positive control, negative control, G-765C single nucleotide polymorphisms
The Reagent Tube for surveying reaction solution is 2ml Reagent Tube.
The above-mentioned kit for being used to detect COX-2 gene pleiomorphism, wherein the G-765C single nucleotide polymorphisms detection
Reaction solution mainly includes: including Taq DNA polymerase, dUTPs, uracil-N-glycosylase (UNG enzyme), specific primer, spy
Needle, MgCL2。
The above-mentioned kit for being used to detect COX-2 gene pleiomorphism, wherein the wild-type positive control is to contain G-
Mitochondria between the wild-type homozygote gene order and its polymorphic detection upstream and downstream primer and primer in the site 765C
The plasmid of DNA sequence dna, saltant type positive control are mutant homozygote gene and its polymorphism containing the site G-765C respectively
Detect the plasmid of the mtdna sequence between upstream and downstream primer and primer.
The above-mentioned kit for being used to detect COX-2 gene pleiomorphism, wherein the freezing box rack is cylindrical structure, cold
Freeze on box frame and be successively arranged first support, second support, third bracket from top to bottom, first support includes the first cylinder, second
Cylinder, the angle between the first cylinder and the second cylinder are 120 °, and second support includes third cylinder, the 4th cylinder, the 5th circle
Column, third cylinder, the 4th cylinder, the angle of the 5th cylinder between any two are 120 °, and third bracket includes the 6th cylinder, the 7th
Cylinder, the 8th cylinder, the 9th cylinder, the 6th cylinder and the 7th cylinder are perpendicular, and the 7th cylinder and the 8th cylinder are perpendicular, and the 8th
Cylinder and the 9th cylinder are perpendicular, and the 9th cylinder and the 6th cylinder are perpendicular；First cylinder, the second cylinder, third cylinder,
4th cylinder, the 5th cylinder, the 6th cylinder, the 7th cylinder, the 8th cylinder, shape, size, the structure of the 9th cylinder are identical.
It is above-mentioned for detecting the kit of COX-2 gene pleiomorphism, wherein the frost box includes first freezing box, the
Two frost boxes, third freeze box, are equipped with first through hole, the second through-hole in the first frost box, and first through hole is circular configuration, and second
Through-hole is string configuration, and first through hole is connected with the second through-hole, and the second frost box includes third through-hole, four-way
Hole, fifth hole, third through-hole are circular configuration, and fourth hole, fifth hole are string configuration, and fourth hole, the 5th are led to
Hole is connected with third through-hole, and the angle between fourth hole and fifth hole is 120 °, and the third frost box includes the 6th
Through-hole, the 7th through-hole, the 8th through-hole, the 9th through-hole, the 6th through-hole are circular configuration, the 7th through-hole, the 8th through-hole, the 9th through-hole
For string configuration, the 7th through-hole, the 8th through-hole, the 9th through-hole are connected with the 6th through-hole, the 7th through-hole, the 8th through-hole,
The angle of nine through-holes between any two is 120 °, second through-hole, fourth hole, fifth hole, the 7th through-hole, the 8th through-hole,
Shape, size, the structure of 9th through-hole are identical, and the first cylinder can be passed through from the second through-hole, and the diameter of first through hole is d1, ice
The diameter for freezing box frame is d2, d1 > d2.
The above-mentioned kit for being used to detect COX-2 gene pleiomorphism, wherein sealing ring, sealing are equipped on the inside of the box cover
The shape and box cover of circle are adapted.
The above-mentioned kit for being used to detect COX-2 gene pleiomorphism, wherein indicia framing is equipped on the outside of the box cover.
The above-mentioned kit for being used to detect COX-2 gene pleiomorphism, wherein the box body, box cover include 3-tier architecture,
It is followed successively by outer layer, insulating layer, internal layer from outside to inside, outer layer is PE material, and insulating layer is PU material, and internal layer is PS material.
Wherein, COX-2 gene G-765C wild primers sequence are as follows:
5 '-GAGGAGAATTTACCGTTCCCG-3 ',
5 '-GAAATACTGTTCTCCGTACCTTCACC-3 '；
Wherein, COX-2 gene G-765C mutant primers sequence are as follows:
5 '-TGAGGAGAATTTACCTGTCCCC-3 ',
5 '-GAAATACTGTTCTCCGTACCTTCACC-3 '；
Wherein, COX-2 gene G-765C wild type detection probe sequence are as follows:
5 '-CTCTCTTTCCAAGAAAC-3 ', 5 end ' markers are FAM, and 3 end ' markers are MGB；
Wherein, COX-2 gene G-765C saltant type detection probe sequence are as follows:
5 '-CTCTCTTTCCAAGAAAC-3 ', 5 end ' markers are JOE, and 3 end ' markers are MGB；
The application method of mentioned reagent box are as follows:
(1) G-765C single nucleotide polymorphisms augmentation detection reaction solution is taken out from kit, room-temperature water bath is thawed, to complete
After full thawing, oscillation is mixed and brief centrifugation is spare；
(2) sample to be examined, preferably EDTA anticoagulated whole blood sample are taken, genomic DNA is extracted；
(3) reaction solution dispenses, and reaction total system is 50uL (PCR reaction solution 35uL+ template 15uL)；
(4) it is loaded, template 15uL is added in the PCR reaction tube for getting out each reaction solution reagent, after covering tightly pipe lid, instantaneously
Low-speed centrifugal, template include: wild-type positive control, saltant type positive control, negative control, sample to be examined DNA；
(5) the preferred ABI7500 fluorescent PCR instrument of PCR amplification: various kinds is filled in the respective sample setting window of instrument software
This title, type, G-765C single nucleotide polymorphisms detect reaction solution corresponding selection FAM and JOE2 sense channel, reference
Fluorescence is set as none.Reaction condition setting: uracil-N-glycosylase is added under the conditions of 37 DEG C and reacts 5 minutes, 95 DEG C pre-
Denaturation 5 minutes, 95 DEG C 15 seconds, 60 DEG C 1 minute, amplified reaction be 40 circulation.
The present invention has the utility model has the advantages that kit of the present invention is suitable for clinical a variety of sample type detections, has specificity
By force, high sensitivity, experimental period is short, easy to operate, safe and non-toxic, equal remarkable advantages at low cost；Kit is not only solved to exist
Thorny problem in transportational process, and during the test, it can quickly take, it is easy to operate；Kit body uses 3 layers
Structure, internal layer are PS material, and safe and non-toxic, high temperature resistant, easy cleaning, insulating layer is PU material, has the function of cryogenic heat transfer, energy
Temperature and humidity is kept to balance for a long time, and shockproof compression-resistant, waterproof and dampproof, outer layer is PE material, and chemical stability is strong, safety and environmental protection,
Wearability is good.
Detailed description of the invention
Fig. 1 is schematic structural view of the invention；
Fig. 2 is that box cover opens schematic diagram；
Fig. 3 is diagrammatic cross-section at A-A in Fig. 2；
Fig. 4 is box body, box cover schematic diagram of internal structure；
Fig. 5 is first support schematic diagram；
Fig. 6 is second support schematic diagram；
Fig. 7 is third support schematic diagram；
Fig. 8 is the first frost box schematic diagram；
Fig. 9 is the second frost box schematic diagram；
Figure 10 is that third freezes box schematic diagram；
In figure: 1- box body, 2- box cover, 3- foam mat, 5- fastener, the first groove of 6-, 7- reagent pore, 8- freeze box
Frame, 9- first support, 10- second support, 11- third bracket, the first cylinder of 12-, the second cylinder of 13-, 14- third cylinder, 15-
4th cylinder, the 5th cylinder of 16-, the 6th cylinder of 17-, the 7th cylinder of 18-, the 8th cylinder of 19-, the 9th cylinder of 20-, 21- first
Box is freezed, 22- second freezes box, and 23- third freezes box, 24- first through hole, the second through-hole of 25-, 26- third through-hole, 27- the
Four through-holes, 28- fifth hole, the 6th through-hole of 29-, the 7th through-hole of 30-, the 8th through-hole of 31-, the 9th through-hole of 32-, 33- sealing ring,
34- indicia framing, 35- outer layer, 36- insulating layer, 37- internal layer.
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described.
For detecting the kit of COX-2 gene pleiomorphism, kit includes box body, box cover, foam mat, freeze box
Frame, frost box, with box body by being hinged and connected, box cover is closed with box body by fastener lid box cover, and the first groove is equipped in box body, cold
Freeze the middle position that box frame is vertically arranged at the first groove, foam mat is located in box body, and foam mat is equipped with multiple reagents
Pore is simultaneously equipped with multiple Reagent Tubes equipped with detection reagent, and Reagent Tube includes the Reagent Tube compareed equipped with wild-type positive, is equipped with
The Reagent Tube of saltant type positive control, the Reagent Tube equipped with negative control are reacted equipped with the detection of G-765C single nucleotide polymorphisms
The Reagent Tube of liquid；Reagent pore annular array on the foam mat, above-mentioned is positive equipped with wild-type positive control, saltant type
Property control, negative control, G-765C single nucleotide polymorphisms detection reaction solution Reagent Tube be 2ml Reagent Tube.
Further, G-765C single nucleotide polymorphisms detection reaction solution mainly includes: including Taq DNA polymerase,
DUTPs, uracil-N-glycosylase (UNG enzyme), specific primer, probe, MgCL2；Wild-type positive control for containing
Line grain between the wild-type homozygote gene order and its polymorphic detection upstream and downstream primer and primer in the site G-765C
The plasmid of body DNA sequence dna, saltant type positive control are the mutant homozygote gene containing the site G-765C and its polymorphic respectively
Property detection upstream and downstream primer and primer between mtdna sequence plasmid.
Further, the freezing box rack is cylindrical structure, be successively arranged on freezing box rack from top to bottom first support,
Second support, third bracket, first support include the first cylinder, the second cylinder, the angle between the first cylinder and the second cylinder
It is 120 °, second support includes third cylinder, the 4th cylinder, the 5th cylinder, and third cylinder, the 4th cylinder, the 5th cylinder are two-by-two
Between angle be 120 °, third bracket include the 6th cylinder, the 7th cylinder, the 8th cylinder, the 9th cylinder, the 6th cylinder with
7th cylinder is perpendicular, and the 7th cylinder and the 8th cylinder are perpendicular, and the 8th cylinder and the 9th cylinder are perpendicular, the 9th cylinder and
Six cylinders are perpendicular；First cylinder, the second cylinder, third cylinder, the 4th cylinder, the 5th cylinder, the 6th cylinder, the 7th circle
Column, the 8th cylinder, shape, size, the structure of the 9th cylinder are identical.
Further, the frost box includes the first frost box, the second frost box, third frost box, and first freezes in box
Equipped with first through hole, the second through-hole, first through hole is circular configuration, and the second through-hole is string configuration, first through hole and second
Through-hole is connected, and the second frost box includes third through-hole, fourth hole, fifth hole, and third through-hole is circular configuration, the
Four through-holes, fifth hole are string configuration, and fourth hole, fifth hole are connected with third through-hole, fourth hole and the 5th
Angle between through-hole is 120 °, and the third frost box includes the 6th through-hole, the 7th through-hole, the 8th through-hole, the 9th through-hole, the
Six through-holes are circular configuration, and the 7th through-hole, the 8th through-hole, the 9th through-hole are string configuration, the 7th through-hole, the 8th through-hole, the
Nine through-holes are connected with the 6th through-hole, and the 7th through-hole, the 8th through-hole, the angle of the 9th through-hole between any two are 120 °, and described
Two through-holes, fourth hole, fifth hole, the 7th through-hole, the 8th through-hole, shape, size, the structure of the 9th through-hole are identical, and first
Cylinder can be passed through from the second through-hole, and the diameter of first through hole is d1, and the diameter of frost box frame is d2, d1 > d2.
Further, sealing ring is equipped on the inside of the box cover, the shape and box cover of sealing ring are adapted；On the outside of the box cover
Equipped with indicia framing；The box body, box cover include 3-tier architecture, are followed successively by outer layer, insulating layer, internal layer, outer layer PE from outside to inside
Material, insulating layer are PU material, and internal layer is PS material.
COX-2 gene G-765C wild primers sequence are as follows:
5 '-GAGGAGAATTTACCGTTCCCG-3 ',
5 '-GAAATACTGTTCTCCGTACCTTCACC-3 '；
COX-2 gene G-765C mutant primers sequence are as follows:
5 '-TGAGGAGAATTTACCTGTCCCC-3 ',
5 '-GAAATACTGTTCTCCGTACCTTCACC-3 '；
COX-2 gene G-765C wild type detection probe sequence are as follows:
5 '-CTCTCTTTCCAAGAAAC-3 ', 5 end ' markers are FAM, and 3 end ' markers are MGB；
COX-2 gene G-765C saltant type detection probe sequence are as follows:
5 '-CTCTCTTTCCAAGAAAC-3 ', 5 end ' markers are JOE, and 3 end ' markers are MGB；
After box body in use, frost box is first put in freezes in refrigerator freezing room, then third frost box sequentially passed through the
One bracket, second support are finally placed in above third bracket, then the second frost box is passed through first support, are finally placed in second
Above bracket, then box is freezed by first and is placed in above first support, then each Reagent Tube is placed in reagent pore, cover box
Lid, fastening.
The application method of mentioned reagent box are as follows:
(1) G-765C single nucleotide polymorphisms augmentation detection reaction solution is taken out from kit, room-temperature water bath is thawed, to complete
After full thawing, oscillation is mixed and brief centrifugation is spare；
(2) sample to be examined, preferably EDTA anticoagulated whole blood sample are taken, genomic DNA is extracted；
(3) reaction solution dispenses, and reaction total system is 50uL (PCR reaction solution 35uL+ template 15uL)；
(4) it is loaded, template 15uL is added in the PCR reaction tube for getting out each reaction solution reagent, after covering tightly pipe lid, instantaneously
Low-speed centrifugal, template include: wild-type positive control, saltant type positive control, negative control, sample to be examined DNA；
(5) the preferred ABI7500 fluorescent PCR instrument of PCR amplification: various kinds is filled in the respective sample setting window of instrument software
This title, type, G-765C single nucleotide polymorphisms detect reaction solution corresponding selection FAM and JOE2 sense channel, reference
Fluorescence is set as none.Reaction condition setting: uracil-N-glycosylase is added under the conditions of 37 DEG C and reacts 5 minutes, 95 DEG C pre-
Denaturation 5 minutes, 95 DEG C 15 seconds, 60 DEG C 1 minute, amplified reaction be 40 circulation.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (8)
1. the kit for detecting COX-2 gene pleiomorphism, which is characterized in that kit includes box body, box cover, sponge liner
Pad, freezing box rack, frost box, by being hinged and connected, box cover is closed with box body by fastener lid for box cover and box body, and box body is interior equipped with the
One groove, freezing box rack are vertically arranged at the middle position of the first groove, and foam mat is located in box body, and foam mat is equipped with
Multiple reagent pores are simultaneously equipped with multiple Reagent Tubes equipped with detection reagent, and Reagent Tube includes the reagent equipped with wild-type positive control
Pipe, the Reagent Tube equipped with saltant type positive control, the Reagent Tube equipped with negative control are examined equipped with G-765C single nucleotide polymorphisms
Survey the Reagent Tube of reaction solution；Reagent pore annular array on the foam mat, above-mentioned compares equipped with wild-type positive, is prominent
It is 2ml Reagent Tube that modification positive control, negative control, G-765C single nucleotide polymorphisms, which detect the Reagent Tube of reaction solution,.
2. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that the G-765C
Single nucleotide polymorphisms detection reaction solution mainly includes: including Taq DNA polymerase, dUTPs, uracil-N-glycosylase (UNG
Enzyme), specific primer, probe, MgCL2。
3. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that the wild type
Positive control be the wild-type homozygote gene order containing the site G-765C and its polymorphic detection upstream and downstream primer and
The plasmid of mtdna sequence between primer, saltant type positive control are that the saltant type containing the site G-765C is homozygous respectively
Subbase because and its mtdna sequence between polymorphic detection upstream and downstream primer and primer plasmid.
4. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that the freeze box
Frame is cylindrical structure, is successively arranged first support, second support, third bracket, first support on freezing box rack from top to bottom
Including the first cylinder, the second cylinder, the angle between the first cylinder and the second cylinder is 120 °, and second support includes third circle
Column, the 4th cylinder, the 5th cylinder, third cylinder, the 4th cylinder, the angle of the 5th cylinder between any two are 120 °, third branch
Frame include the 6th cylinder, the 7th cylinder, the 8th cylinder, the 9th cylinder, the 6th cylinder and the 7th cylinder are perpendicular, the 7th cylinder and
8th cylinder is perpendicular, and the 8th cylinder and the 9th cylinder are perpendicular, and the 9th cylinder and the 6th cylinder are perpendicular；First circle
Column, the second cylinder, third cylinder, the 4th cylinder, the 5th cylinder, the 6th cylinder, the 7th cylinder, the 8th cylinder, the 9th cylinder
Shape, size, structure are identical.
5. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that the frost box
Box is freezed including the first frost box, the second frost box, third, is equipped with first through hole, the second through-hole in the first frost box, first is logical
Hole is circular configuration, and the second through-hole is string configuration, and first through hole is connected with the second through-hole, and the second frost box includes
Third through-hole, fourth hole, fifth hole, third through-hole are circular configuration, and fourth hole, fifth hole are string configuration,
Fourth hole, fifth hole are connected with third through-hole, and the angle between fourth hole and fifth hole is 120 °, the third
Freezing box includes the 6th through-hole, the 7th through-hole, the 8th through-hole, the 9th through-hole, and the 6th through-hole is circular configuration, the 7th through-hole, the
Eight through-holes, the 9th through-hole are string configuration, and the 7th through-hole, the 8th through-hole, the 9th through-hole are connected with the 6th through-hole, and the 7th is logical
Hole, the 8th through-hole, the angle of the 9th through-hole between any two are 120 °, second through-hole, fourth hole, fifth hole, the 7th
Through-hole, the 8th through-hole, shape, size, the structure of the 9th through-hole are identical, and the first cylinder can be passed through from the second through-hole, and first is logical
The diameter in hole is d1, and the diameter of frost box frame is d2, d1 > d2.
6. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that in the box cover
Side is equipped with sealing ring, and the shape and box cover of sealing ring are adapted.
7. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that outside the box cover
Side is equipped with indicia framing.
8. as described in claim 1 for detecting the kit of COX-2 gene pleiomorphism, which is characterized in that the box body, box
Lid includes 3-tier architecture, is followed successively by outer layer, insulating layer, internal layer from outside to inside, and outer layer is PE material, and insulating layer is PU material, interior
Layer is PS material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811191253.6A CN108949532A (en) | 2018-10-12 | 2018-10-12 | For detecting the kit of COX-2 gene pleiomorphism |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811191253.6A CN108949532A (en) | 2018-10-12 | 2018-10-12 | For detecting the kit of COX-2 gene pleiomorphism |
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| CN108949532A true CN108949532A (en) | 2018-12-07 |
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| CN201811191253.6A Pending CN108949532A (en) | 2018-10-12 | 2018-10-12 | For detecting the kit of COX-2 gene pleiomorphism |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110895283A (en) * | 2019-12-10 | 2020-03-20 | 宁波奥丞生物科技有限公司 | High-sensitivity D-dimer detection kit and use method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080220420A1 (en) * | 2004-11-19 | 2008-09-11 | Shimadzu Corporation | Method of Detecting Gene Polymorphism, Method of Diagnosing, Apparatus Therefor, and Test Reagent Kit |
| CN105671036A (en) * | 2015-09-10 | 2016-06-15 | 南京仁天生物科技有限公司 | Kit for detecting human COX-2 gene polymorphism and application thereof |
| CN205347424U (en) * | 2016-01-27 | 2016-06-29 | 上海睿玻生物科技有限公司 | TPMT gene polymorphism detection kit |
| CN106834434A (en) * | 2016-12-06 | 2017-06-13 | 武汉海吉力生物科技有限公司 | For detecting COX 1, the nucleic acid of COX 2 and GPIIIa gene pleiomorphisms, kit and method |
| CN206359521U (en) * | 2016-12-26 | 2017-07-28 | 上海新培晶医学检验所有限公司 | A kind of kit for being used to detect Dubin Johnson syndrome gene mutations |
| CN206692651U (en) * | 2017-03-21 | 2017-12-01 | 北京军科基因医学研究院有限公司 | A kind of children's Loratadine genetic polymorphism detection kit |
| CN206814773U (en) * | 2017-04-17 | 2017-12-29 | 新疆出入境检验检疫局检验检疫技术中心 | Detection GMOs kit |
| CN108342310A (en) * | 2018-03-19 | 2018-07-31 | 吴娜 | A kind of mutation of cancer gene and gene spread detection kit |
-
2018
- 2018-10-12 CN CN201811191253.6A patent/CN108949532A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080220420A1 (en) * | 2004-11-19 | 2008-09-11 | Shimadzu Corporation | Method of Detecting Gene Polymorphism, Method of Diagnosing, Apparatus Therefor, and Test Reagent Kit |
| CN105671036A (en) * | 2015-09-10 | 2016-06-15 | 南京仁天生物科技有限公司 | Kit for detecting human COX-2 gene polymorphism and application thereof |
| CN205347424U (en) * | 2016-01-27 | 2016-06-29 | 上海睿玻生物科技有限公司 | TPMT gene polymorphism detection kit |
| CN106834434A (en) * | 2016-12-06 | 2017-06-13 | 武汉海吉力生物科技有限公司 | For detecting COX 1, the nucleic acid of COX 2 and GPIIIa gene pleiomorphisms, kit and method |
| CN206359521U (en) * | 2016-12-26 | 2017-07-28 | 上海新培晶医学检验所有限公司 | A kind of kit for being used to detect Dubin Johnson syndrome gene mutations |
| CN206692651U (en) * | 2017-03-21 | 2017-12-01 | 北京军科基因医学研究院有限公司 | A kind of children's Loratadine genetic polymorphism detection kit |
| CN206814773U (en) * | 2017-04-17 | 2017-12-29 | 新疆出入境检验检疫局检验检疫技术中心 | Detection GMOs kit |
| CN108342310A (en) * | 2018-03-19 | 2018-07-31 | 吴娜 | A kind of mutation of cancer gene and gene spread detection kit |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110895283A (en) * | 2019-12-10 | 2020-03-20 | 宁波奥丞生物科技有限公司 | High-sensitivity D-dimer detection kit and use method thereof |
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