CN106018792A - Immunochromatography kit for testing of stomach function and preparation method of immunochromatography kit - Google Patents
Immunochromatography kit for testing of stomach function and preparation method of immunochromatography kit Download PDFInfo
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- CN106018792A CN106018792A CN201610327064.1A CN201610327064A CN106018792A CN 106018792 A CN106018792 A CN 106018792A CN 201610327064 A CN201610327064 A CN 201610327064A CN 106018792 A CN106018792 A CN 106018792A
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- pepsinogen
- quantum dot
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- antibody
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- 238000012360 testing method Methods 0.000 title claims abstract description 43
- 230000004206 stomach function Effects 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 238000003317 immunochromatography Methods 0.000 title abstract description 5
- 238000001514 detection method Methods 0.000 claims abstract description 121
- 108010047320 Pepsinogen A Proteins 0.000 claims abstract description 109
- 239000002096 quantum dot Substances 0.000 claims abstract description 95
- 239000000427 antigen Substances 0.000 claims abstract description 44
- 102000036639 antigens Human genes 0.000 claims abstract description 44
- 108091007433 antigens Proteins 0.000 claims abstract description 44
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 33
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 33
- 238000000034 method Methods 0.000 claims abstract description 21
- 239000000203 mixture Substances 0.000 claims abstract description 20
- 239000004033 plastic Substances 0.000 claims abstract description 10
- 101150061086 ureB gene Proteins 0.000 claims description 44
- 239000004816 latex Substances 0.000 claims description 37
- 229920000126 latex Polymers 0.000 claims description 37
- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 claims description 31
- 239000003365 glass fiber Substances 0.000 claims description 29
- 239000000725 suspension Substances 0.000 claims description 22
- 150000001875 compounds Chemical class 0.000 claims description 19
- 239000002245 particle Substances 0.000 claims description 19
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical class O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 claims description 15
- 238000002156 mixing Methods 0.000 claims description 12
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- 238000000746 purification Methods 0.000 claims description 11
- 241001494479 Pecora Species 0.000 claims description 10
- 238000012986 modification Methods 0.000 claims description 9
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- 238000005576 amination reaction Methods 0.000 claims description 4
- UHYPYGJEEGLRJD-UHFFFAOYSA-N cadmium(2+);selenium(2-) Chemical group [Se-2].[Cd+2] UHYPYGJEEGLRJD-UHFFFAOYSA-N 0.000 claims description 4
- 238000011017 operating method Methods 0.000 claims description 4
- WUPHOULIZUERAE-UHFFFAOYSA-N 3-(oxolan-2-yl)propanoic acid Chemical compound OC(=O)CCC1CCCO1 WUPHOULIZUERAE-UHFFFAOYSA-N 0.000 claims description 3
- 229920002160 Celluloid Polymers 0.000 claims description 3
- 229940116367 cadmium sulfide Drugs 0.000 claims description 3
- 229910052980 cadmium sulfide Inorganic materials 0.000 claims description 3
- 239000007921 spray Substances 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 2
- 241000590002 Helicobacter pylori Species 0.000 abstract description 10
- 229940037467 helicobacter pylori Drugs 0.000 abstract description 10
- 108010046334 Urease Proteins 0.000 abstract description 9
- 108090001072 Gastricsin Proteins 0.000 abstract description 6
- 102000034255 Pepsinogen C Human genes 0.000 abstract description 6
- 239000012528 membrane Substances 0.000 abstract description 6
- 229920002678 cellulose Polymers 0.000 abstract description 3
- 239000001913 cellulose Substances 0.000 abstract description 3
- 239000011521 glass Substances 0.000 abstract description 3
- 230000002745 absorbent Effects 0.000 abstract 1
- 239000002250 absorbent Substances 0.000 abstract 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 25
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- 229940111202 pepsin Drugs 0.000 description 10
- 238000001556 precipitation Methods 0.000 description 10
- 108090000790 Enzymes Proteins 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 8
- 210000002784 stomach Anatomy 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 239000013618 particulate matter Substances 0.000 description 7
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 6
- 238000004587 chromatography analysis Methods 0.000 description 6
- 238000002372 labelling Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000007987 MES buffer Substances 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 210000001187 pylorus Anatomy 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 108010062466 Enzyme Precursors Proteins 0.000 description 3
- 102000010911 Enzyme Precursors Human genes 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 235000013877 carbamide Nutrition 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- 210000003097 mucus Anatomy 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 2
- 241000207199 Citrus Species 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
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- 238000005119 centrifugation Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 235000020971 citrus fruits Nutrition 0.000 description 2
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- 239000008187 granular material Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 230000011987 methylation Effects 0.000 description 2
- 238000007069 methylation reaction Methods 0.000 description 2
- 238000013139 quantization Methods 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical group Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- 208000004300 Atrophic Gastritis Diseases 0.000 description 1
- 206010003694 Atrophy Diseases 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 208000007882 Gastritis Diseases 0.000 description 1
- 208000036495 Gastritis atrophic Diseases 0.000 description 1
- 229920000715 Mucilage Polymers 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000008469 Peptic Ulcer Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
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- 230000000844 anti-bacterial effect Effects 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- -1 centrifugation Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007813 chromatographic assay Methods 0.000 description 1
- 208000016644 chronic atrophic gastritis Diseases 0.000 description 1
- 208000023652 chronic gastritis Diseases 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000001198 duodenum Anatomy 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 229940127121 immunoconjugate Drugs 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 201000011591 microinvasive gastric cancer Diseases 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 208000011906 peptic ulcer disease Diseases 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 210000004203 pyloric antrum Anatomy 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
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- 230000001105 regulatory effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of immunochromatography detection and particularly discloses a kit for testing of the stomach function and a preparation method of the kit. The kit comprises a plastic board, a glass cellulose membrane, a nitrocellulose membrane, a sample pad and a water absorbent pad, wherein the glass cellulose membrane is coated with a quantum-dot-labeled mixture of pepsinogen I antibodies, pepsinogen II antibodies and helicobacter pylori urease antigens; a detection line 1, a detection line 2, a detection line 3 and a control line are arranged on the nitrocellulose membrane, the detection line 1 is coated with pepsinogen I secondary antibodies, the detection line 2 is coated with pepsinogen II secondary antibodies, the detection line 3 is coated with helicobacter pylori urease antigens, and the control line is coated with goat-anti-mouse lgG. According to the kit, fast and sensitive combined detection of pepsinogen I, pepsinogen II and helicobacter pylori urease antibodies is realized with the fluorescence immunochromatography method by means of the double-antibody/antigen sandwich principle in combination with the fluorescence characteristic of quantum dots.
Description
Technical field
The present invention relates to immuno-chromatographic assay technology field, be specifically related to a kind of stomach Function detection immunochromatography
Test kit and preparation method thereof.
Background technology
Pepsic inactive precursor in the original gastric juice of pepsin, by chief cell and the mucilage cell of body of stomach
Secretion, major part is secreted into gastral cavity, can be detected in blood on a small quantity.According to its biochemical property and immunity
Originality is divided into 2 subgroups, and the immunogenicity of 1-5 component is identical, referred to as pepsinogen I, mainly by
The chief cell of fundic gland and the secretion of mucus neck cell；Component 6 and 7 is referred to as pepsinogen I I, except by stomach
Outside the chief cell of end gland and the secretion of mucus neck cell, the mucus neck cell of the pyloric gland of cardiac gland and gastric antrum and
Duodenum epimere also can produce pepsinogen I I.Pepsinogen I and pepsin in serum and blood plasma
The former reduction of II ratio is proportionate with the order of severity of corpus atrophy, in, severe atrophic gastritis time, its
Ratio is ＜ 0.3, and the sickness rate of its gastric cancer dramatically increases, therefore, and simultaneous determination pepsinogen I core stomach egg
White proenzyme II ratio can play the effect of early gastric cancer indicating risk.It addition, stomach Helicobacter pylori is existence
The pathogen of human gastric mucosal, it is one of the weight virulence factor of chronic gastritis, peptic ulcer, gastric cancer,
Being the people's gastric antibacterial that uniquely can produce a large amount of uremic enzyme, therefore detection stomach Helicobacter pylori antibody is to stomach
The judgement of function stomach function regulating cancer morbidity has tell-tale effect.
The method predominantly detected for pepsinogen and HP ureB antibody at present is joined by enzyme
Immunoabsorption, CLIA and fluorescence immune chromatography method.Enzyme linked immunosorbent assay detection is accurate,
Can accurate quantitative analysis, there is the highest sensitivity, but detection sample needs sample-adding, temperature to bathe, washs, develops the color,
The steps such as termination, operation sequence is loaded down with trivial details, the longest, sample from process to it is concluded that need at least 40 minutes,
Be not suitable for large-scale health check-up examination；Sample detection needs to wash trigger and microplate reader, inconvenient to carry, thus not
Can detect in patient family or emergency tender；It addition, its sensitivity, specificity are not high enough, to low dense
The accuracy of degree sample detection is the highest.And CLIA, although degree of accuracy is high, and detection speed is fast,
But detection needs expensive chemical illumination immunity analysis instrument, it is desirable to specific analysis room, and its reagent cost is also
Higher, it is impossible to be widely used in wish and the medical institutions of relatively basic unit.Fluorescence immune chromatography method is compared enzyme connection and is exempted from
Epidemic disease absorption method and CLIA, detection sensitivity is high, accuracy is high, easy and simple to handle, low cost,
Thus make it have bigger industry in the context of detection of pepsinogen and HP ureB antibody
It is worth, but either uses which kind of detection method at present, be to pepsinogen I, pepsin mostly
Former II and HP ureB antibody individually detect, be also not carried out to pepsinogen I,
Pepsinogen I I and a step joint-detection of three kinds of materials of HP ureB antibody.
Summary of the invention
In order to overcome the defect of prior art, it is an object of the invention to provide a kind of highly sensitive, accuracy is high,
Easy and simple to handle, the stomach Function detection test kit of low cost, it is achieved to pepsinogen I, pepsinogen
Joint-detection while II and three kinds of compositions of HP ureB antibody.
The present invention also aims to provide the preparation method of a kind of stomach Function detection test kit.
In order to realize object above, the technical solution adopted in the present invention is:
A kind of stomach Function detection test kit, the glass fibre element film including plastic plate, being arranged on plastic plate,
Being arranged on the nitrocellulose filter on glass fibre element film, on described nitrocellulose filter, one end is provided with sample pad,
The corresponding other end is provided with adsorptive pads, and described glass fibre element film is coated with quantum dot-labeled pepsin
Former I antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled helicobacter pylori carbamide
The mixture of enzyme antigen；Detection line 1, detection line 2, detection line 3 it is provided with on described nitrocellulose filter
And control line, described detection line 1 is coated with pepsinogen I two and resists, described detection line 2 is coated with
Pepsinogen I I bis-resists, and described detection line 3 is coated with HP ureB antigen, described control
Sheep anti mouse lgG it is coated with on line processed.
Coated quantum dot-labeled pepsinogen I antibody, quantum dot-labeled stomach on described glass fibre
Quantum dot mark in the mixture of pepsinogen II antibody and quantum dot-labeled HP ureB antigen
Pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody and the quantum dot-labeled pylorus of note
The mass ratio of pylori urease antigen is 1:1:1.
Described quantum dot is cadmium selenide or cadmiumsulfide quantum dot.
Between the detection line 1 and the detection line 2 that arrange on described nitrocellulose filter, detect line 2 and detection line
Interval between 3, between detection line 1 and control line is all not less than 5mm.
The preparation method of above-mentioned stomach Function detection test kit, including following operating procedure:
1) raw-material preparation:
A: prepare glass fibre element film: by quantum dot-labeled pepsinogen I antibody, quantum dot-labeled
Pepsinogen I I antibody, quantum dot-labeled HP ureB antigen mixing be coated on glass
On cellulose membrane, it is dried, standby；
B: prepare nitrocellulose filter: take nitrocellulose filter, interval mark off detection line 1, detection line 2,
Detection line 3 and control line, then by anti-for pepsinogen I two, anti-, the pylorus spiral of pepsinogen I I bis-
Bacillus urease antigen, sheep anti mouse lgG be coated on respectively nitrocellulose filter detection line 1, detection line 2,
On detection line 3 and control line, the nitrocellulose filter after being coated afterwards is put in confining liquid and is closed, and is dried,
Standby；
2) assemble: on plastic plate, place step 1) the glass fibre prime modulus prepared, then in step 1)
Step 1 is placed on the glass fibre element film of preparation) nitrocellulose filter prepared, then at celluloid
One end place sample pad, the corresponding other end places adsorptive pads, is dried, i.e. completes.
Step A also includes first preparing quantum dot-labeled pepsinogen I antibody, quantum dot-labeled
Pepsinogen I I antibody and the mixture of quantum dot-labeled HP ureB antigen, specifically side
Method includes following operating procedure:
A: add modified cyclodextrin in latex particle suspension, mix 24 hours, purification, obtain modified ring
Dextrin-latex compound suspension；
B: add quantum dot in modified cyclodextrin-latex compound suspension prepared by step a, mix 24
Hour, purification, obtain quantum dot-modified cyclodextrin-latex compound suspension；
C: add activator in quantum dot-modified cyclodextrin-latex compound suspension prepared by step b,
Activate 15 minutes, purification, add pepsinogen I antibody afterwards, after mixing 2～4 hours, heavyly centrifugal
Shallow lake composition granule, by resuspended for precipitate particles confining liquid, room temperature mixes 1 hour, is purified the most again,
Obtain quantum dot-labeled pepsinogen I antibody；
D: according to the method that above-mentioned steps a～c are same prepare quantum dot-labeled pepsinogen I I antibody and
Quantum dot-labeled HP ureB antigen, then resists quantum dot-labeled pepsinogen I
Body, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB antigen
Mixing, standby；
Described modified cyclodextrin is carboxymethyl-modification cyclodextrin or amination modified cyclodextrin.
Described activator is 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride.
Described in step c, quantum dot-modified cyclodextrin-latex compound suspension with the mass ratio of activator is
1:10.
Confining liquid used in step B is bovine serum albumin solution.
Confining liquid used in step c is bovine serum albumin solution.
Modified cyclodextrin described in step a is 20:1 with the mass ratio of latex particle；Measure described in step b
Son point is 10:1 with the mass ratio of cyclodextrin-latex compounds；Pepsinogen I antibody described in step c
It is 1:20 with the mass ratio of quantum dot-modified cyclodextrin-latex compound.
In step a, latex particle carries out purification process before using, described purification process method particularly includes: take
Latex particle adds in the first buffer, centrifugation, supernatant discarded, then by the first resuspended precipitation of buffer
Composition granule, the most again centrifugation, supernatant discarded, then with the first resuspended precipitate particles of buffer, 4 DEG C of guarantors
Deposit standby.
Above-mentioned latex particle after purification takes latex particle in use, uses the second buffer by latex particle
Being diluted to concentration is 10mg/ml.
Above-mentioned steps b also includes first by suspended for the modified cyclodextrin-latex compound of preparation in step a
Liquid, use the second buffer to be diluted to suspension that concentration is 10mg/ml, then add quantum dot.
Above-mentioned steps c also includes first be combined by the quantum dot-modified cyclodextrin-latex of preparation in step b
Thing suspension, use the second buffer to be diluted to suspension that concentration is 10mg/ml, then add activation
Agent activates.
It is purified after closing described in step c method particularly includes: centrifugal 15min under the conditions of 8000rpm,
Supernatant discarded, precipitation is redissolved in dispersion liquid.
By quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I in step A
The mixing of antibody, quantum dot-labeled HP ureB antigen is coated on the tool on glass fibre element film
Body method is: the most quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I
Antibody and quantum dot-labeled HP ureB antigen mixture even application are in glass fibre element film
On.
Purification described in step a and step b method particularly includes: centrifugal under the conditions of 8000rpm, discards
Clearly, precipitation carries out resuspended with the first buffer, centrifugal under the conditions of 8000rpm afterwards, and supernatant discarded is heavy
Shallow lake is redissolved in the first buffer.
Purification after activation in step c method particularly includes: centrifugal supernatant discarded, precipitation under the conditions of 8000rpm
Carry out resuspended with the first buffer.
By anti-for pepsinogen I two, anti-, the HP ureB of pepsinogen I I bis-in step B
Antigen, sheep anti mouse lgG are coated on the detection line 1 of nitrocellulose filter, detection line 2, detection line 3 respectively
With on control line method particularly includes: take concentration respectively and be that the pepsinogen I two of 1mg/ml is anti-, stomach egg
White proenzyme II bis-is anti-, HP ureB antigen and sheep anti-mouse igg, with 1ul/cm, the 10cm/ second
Speed sprays on the detection line 1 of cellulose nitrate, detection line 2, detection line 3 and control line respectively.
Step 1) and step 2) described in be dried be 37 DEG C be dried 3～5 hours.
Above-mentioned first buffer is the PB buffer of 0.02M, pH7.4.Above-mentioned second buffer is 0.02M,
The MES buffer of pH6.1.
Testing sample in use, is added in the sample pad of test kit by above-mentioned stomach Function detection test kit,
The pepsinogen I antibody of the quantization labelling on glass fibre element film, quantum dot-labeled pepsinogen
II antibody, quantum dot-labeled the most specific the capturing of HP ureB antigen mixture are treated
Pepsinogen I, pepsinogen I I and HP ureB antibody in test sample product, then exists
When reaching to detect line 3, quantum dot-labeled HP ureB antigen-pylorus spiral Citrus urease
Antibody conjugates pylorus spiral Citrus urease antigenic specificity on detection line 3 is combined,；Successively inspection
During survey line 2, quantum dot-labeled pepsinogen I I antibody-pepsinogen I I conjugate and detection line 2
On pepsinogen I I bis-resist specific binding；When detecting line 1, quantum dot-labeled pepsinogen
I antibody-pepsinogen I conjugate resists specific binding with the pepsinogen I two on detection line 1；It
The fluorescent effect of the quantum dot by combining on detection detection line 1, detection line 2 and detection line 3 afterwards, quantitatively
Resist with qualitatively judging pepsinogen I, pepsinogen I I and HP ureB in testing sample
Body.
Stomach Function detection test kit of the present invention, with pepsinogen I antibody, the quantum dot of quantization labelling
The pepsinogen I I antibody of labelling, quantum dot-labeled HP ureB antigen mixture are coated
On glass fibre element film, as detection probe, utilize the glimmering of double antibody/antigen sandwich principle incorporating quantum point
Light characteristic achieves and uses the method for fluorescence immune chromatography to pepsinogen I, pepsinogen I I and pylorus
Pylori urease antibody carries out quick, special, easy, sensitive joint-detection.Stomach function of the present invention
Detection test kit detection line is low, and detection sensitivity is high.
The preparation method of stomach Function detection test kit of the present invention, easy and simple to handle, it is easy to control, be suitable to industry
Change popularization and application.
It is further preferred that in preparation method of the present invention, innovate quantum dot-labeled technology, use carboxy methylation
Or amination modified cyclodextrin modified quantum dot, then by pepsinogen I antibody, pepsinogen I I
Antibody, HP ureB antigen are bonded respectively to carboxymethyl group or the amino base of modified cyclodextrin
In group, it is achieved prepare quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsin
Former II antibody, quantum dot-labeled HP ureB antigen.
Accompanying drawing explanation
Fig. 1 is test kit detection pepsinogen Cgene standard sample luminous value and the reality using embodiment 1 preparation
The standard curve of corresponding relation between the concentration of border；
Fig. 2 is test kit detection pepsinogen II standard sample luminous value and the reality using embodiment 1 preparation
The standard curve of corresponding relation between the concentration of border；
Fig. 3 is the test kit detection HP ureB antigen standard sample using embodiment 1 preparation
The standard curve of corresponding relation between luminous value and actual concentrations.
Detailed description of the invention
Below by specific embodiment, technical scheme is described in detail.
Embodiment 1
The present embodiment stomach Function detection test kit, including plastic plate, the glass fibre that is arranged on plastic plate
Element film, the nitrocellulose filter being arranged on glass fibre element film, on described nitrocellulose filter, one end is provided with
Sample pad, the corresponding other end is provided with adsorptive pads, and it is 1:1 that described glass fibre element film is coated with mass ratio:
The quantum dot-labeled pepsinogen I antibody of 1, quantum dot-labeled pepsinogen I I antibody and quantum
The mixture of the HP ureB antigen of some labelling；It is provided with detection on described nitrocellulose filter
Line 1, detection line 2, detection line 3 and control line, described detection line 1 is coated with pepsinogen I two and resists,
It is coated with pepsinogen I I bis-on described detection line 2 to resist, described detection line 3 is coated with H. pylori
Bacterium urease antigen, described control line is coated with sheep anti mouse lgG；Described quantum dot is CdSe quantum dots.
The preparation method of the present embodiment stomach Function detection test kit, concrete operation step is:
1) glass fibre element film is prepared:
A: take the unmarked latex particle of 1ml and add 10ml, in 0.02M, pH7.4PB buffer；Centrifugal
Machine 10000rpm is centrifuged 20min；Remove supernatant, then with 10ml, 0.02M, pH7.4PB buffer
Resuspended precipitation particulate matter；Centrifuge 10000rpm is centrifuged 20min the most again, removes supernatant and uses 5ml
0.02M, pH7.4, PB buffer resuspended precipitation particulate matter, 4 DEG C of preservations are stand-by；
B: take the MES buffer of latex particle 0.02M, pH6.1 prepared by step a by latex particle
It is diluted to 10mg/ml, takes the latex particle suspension after 0.05ml dilution and add 10mg carboxymethyl-modification
Cyclodextrin, after mixing 24 hours, centrifuge 8000rpm is centrifuged, supernatant discarded, and with 0.02M, pH7.4
PB buffer resuspended precipitation particulate matter, recentrifuge machine 8000rpm is centrifuged afterwards, and supernatant discarded is heavy
Shallow lake particulate matter redissolves in the PB buffer of 0.02M, pH7.4, obtains carboxymethyl-modification cyclodextrin-latex
Complex suspension；
C: take carboxymethyl-modification cyclodextrin-latex compound suspension prepared by step b, with 0.02M,
The MES buffer of pH6.1 is diluted to 10mg/ml, takes the carboxymethyl-modification ring after 20ml dilution
Dextrin-latex is combined suspension, adds 20mg CdSe quantum dots, mixes 24 hours, centrifuge 8000rpm
Centrifugal, supernatant discarded, and with the PB buffer resuspended precipitation particulate matter of 0.02M, pH7.4, the most again
Secondary centrifuge 8000rpm is centrifuged, supernatant discarded, and precipitation particulate matter redissolves in the PB of 0.02M, pH7.4
In buffer, obtain quantum dot-carboxymethyl-modification cyclodextrin-latex compound suspension；
D: take quantum dot-carboxymethyl-modification cyclodextrin-latex compound suspension prepared by step c, uses
The MES buffer of 0.02M, pH6.1 is diluted to 10mg/ml, then according to quantum dot-modification ring is stuck with paste
Essence-latex compound is 1:10 with the mass ratio of activator, adds 1-(3-dimethylamino-propyl)-3-ethyl carbon
Diimmonium salt hydrochlorate, after activating 15 minutes, 8000rpm is centrifuged, supernatant discarded, and with 0.02M, pH7.4
The resuspended precipitate particles of PB buffer, afterwards according to pepsinogen I antibody: quantum dot-carboxy methylation
The ratio of modified cyclodextrin-latex compound=1:20 adds pepsinogen I antibody, after mixing 3 hours,
8000rpm is centrifuged 15 minutes, by resuspended for precipitate particles confining liquid, and room temperature mixing 1 hour, afterwards
8000rpm is centrifuged 15min, and precipitation particulate matter redissolves in dispersion liquid, obtains quantum dot-labeled pepsin
Former I antibody；
E: according to the method that above-mentioned steps a～c are same prepare quantum dot-labeled pepsinogen I I antibody and
Quantum dot-labeled HP ureB antigen, then resists quantum dot-labeled pepsinogen I
Body, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB antigen
Mixing, is sprayed on glass fibre prime modulus, and 37 DEG C are dried 4 hours, obtain quantum dot-labeled pepsin
Former I antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled helicobacter pylori carbamide
The glass fibre element film of enzyme antigen mixture labelling；
2) prepare nitrocellulose filter: take nitrocellulose filter, interval 6mm divide successively detection line 3,
Detection line 2, detection line 1 and control line, take afterwards concentration be the pepsinogen I two of 1mg/ml anti-,
Pepsinogen I I bis-anti-, HP ureB antigen, sheep anti mouse igG, with 1ul/cm, 10cm/
The speed of second sprays on the detection line 1 of nitrocellulose membrane, detection line 2, detection line 3 and control line respectively,
Being put into by nitrocellulose filter afterwards in bovine serum albumin solution and close, 37 DEG C are dried 3 hours, obtain institute
The nitrocellulose filter stated；
3) assemble: on plastic plate, place step 1) the glass fibre prime modulus prepared, in step 1) prepare
Glass fibre element film on place step 2) nitrocellulose filter prepared, then at the one of celluloid
End places sample pad, and the corresponding other end places adsorptive pads, and 37 DEG C are dried 5 hours, i.e. complete.
Embodiment 2
The present embodiment stomach Function detection test kit, the place different from embodiment is, the quantum dot of employing
For cadmiumsulfide quantum dot, the modified cyclodextrin of employing is amination dryness cyclodextrin, prepares pepsinogen I
Antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB are anti-
During former, the quantum dot-modified cyclodextrin-latex suspension of activation adds the mixed of antibody or antigen
The even time is 2 hours, and other are with embodiment 1.
Embodiment 3
The present embodiment stomach Function detection test kit, the place different from embodiment is, prepares pepsin
Former I antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled helicobacter pylori carbamide
During enzyme antigen, the quantum dot-modified cyclodextrin-latex suspension of activation adds antibody or antigen
Mixing time be 4 hours, other are with embodiment 1.
Test example:
Pepsinogen I is diluted to 7.8125ug/L, 15.625ug/L, 31.25ug/L, 62.5ug/L,
125ug/L、250ug/L、500ug/L；Pepsinogen I I be diluted to 3.90625ug/L, 7.8125ug/L,
15.625ug/L、31.25ug/L、62.5ug/L、125ug/L、250ug/L；HP ureB
Antibody be diluted to 1.5625ug/L, 3.125ug/L, 6.25ug/L, 12.5ug/L, 25ug/L, 50ug/L,
100ug/L is as standard solution, and prepares blank solution simultaneously.
The standard substance 100ul taking each concentration respectively is added separately to the sample pad of test kit prepared by embodiment 1
On, (Nanjing Mei Ningkang is the most biological after 5min, the test kit that colour developing completes to be put into fluorescence immune chromatography readout instrument
Science and Technology Ltd.) weight, reads the corresponding luminous value detecting line, and records the detection of each concentration standard sample
Corresponding relation between luminous value and actual concentrations, draws standard curve, as shown in FIG. 1 to 3, pepsin
The standard curve regression equation of proenzyme I is: y=-0.00003x2+ 0.03545x+0.11366, R2=0.99967；
The standard curve regression equation of pepsinogen I I is: y=-0.00013x2+ 0.07164x-0.03524,
R2=0.99983；The standard curve regression equation of helicobacter pylori uremic enzyme antibody: y=-0.00039
x2+ 0.08853x+0.01878, R2=0.99965, above-mentioned standard curve understand test kit prepared by the present invention
Be limited to 2 μ g/L for the lowest detection of pepsinogen I, the lowest detection of pepsinogen I I is limited to 1 μ
G/L, the lowest detection of helicobacter pylori uremic enzyme antibody are limited to 0.5 μ g/L.
It addition, take in the test kit that blank solution joins embodiment 1 preparation, it is not detected by fluorescent effect.
Take the clinical sample that 100 examples are random, use the test kit of the embodiment of the present invention 1 preparation to pepsin
The concentration of former I, pepsinogen I I and HP ureB antibody carries out joint-detection.Detection side
Method is: 100ul testing sample (50ul sample+50ul sample diluting liquid) instills in test kit sample pad, 5min
After the test strips of standard substance that colour developing is completed and testing sample put into fluorescence immune chromatography readout instrument (Nanjing be beautiful
Ning Kangcheng bio tech ltd) in, read the luminous value of T/C line, and record, by testing sample T/C
Value is brought in standard curve, obtains pepsinogen I, pepsinogen I I and HP ureB
Antibody concentration result, as shown in table 1 below:
Table 1
As can be seen here, this method can detect pepsinogen with direct quantitative, is not required to professional training, operation side
Just, quickly, 20min can obtain result, is suitable for promoting at basic hospital and MEC and using.
Last it is noted that above example is only in order to illustrate technical scheme, rather than it is limited
System；Although the present invention being described in detail with reference to previous embodiment, those of ordinary skill in the art
It is understood that the technical scheme described in foregoing embodiments still can be modified by it, or to it
Middle part technical characteristic carries out equivalent；And these amendments or replacement, do not make appropriate technical solution
Essence departs from the spirit and scope of various embodiments of the present invention technical scheme.
Claims (10)
1. a stomach Function detection test kit, the glass fibre element film including plastic plate, being arranged on plastic plate, the nitrocellulose filter being arranged on glass fibre element film, on described nitrocellulose filter, one end is provided with sample pad, the corresponding other end is provided with adsorptive pads, it is characterized in that, described glass fibre element film is coated with quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody and the mixture of quantum dot-labeled HP ureB antigen；Detection line 1, detection line 2, detection line 3 and control line it is provided with on described nitrocellulose filter, it is coated with pepsinogen I two on described detection line 1 to resist, it is coated with pepsinogen I I bis-on described detection line 2 to resist, it is coated with HP ureB antigen on described detection line 3, described control line is coated with sheep anti mouse lgG.
2. stomach Function detection test kit as claimed in claim 1, it is characterized in that, the mass ratio of pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB antigen that coated quantum dot-labeled pepsinogen I antibody on described glass fibre, quantum dot-labeled pepsinogen I I antibody are quantum dot-labeled with in the mixture of quantum dot-labeled HP ureB antigen is 1:1:1.
3. stomach Function detection test kit as claimed in claim 1, it is characterised in that described quantum dot is cadmium selenide or cadmiumsulfide quantum dot.
4. the stomach Function detection test kit as described in any one of claims 1 to 3, it is characterized in that, the interval between detection line 1 and detection line 2 that described nitrocellulose filter is arranged, between detection line 2 and detection line 3, between detection line 1 and control line is all not less than 5mm.
5. the preparation method of a stomach Function detection test kit as claimed in claim 1, it is characterised in that include following operating procedure:
1) raw-material preparation:
A: prepare glass fibre element film: quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody, the mixing of quantum dot-labeled HP ureB antigen are coated on glass fibre element film, are dried, standby；
B: prepare nitrocellulose filter: take nitrocellulose filter, interval marks off detection line 1, detection line 2, detection line 3 and control line, then by detection line 1 anti-for pepsinogen I two, that pepsinogen I I bis-is anti-, HP ureB antigen, sheep anti mouse lgG are coated on nitrocellulose filter respectively, detection line 2, detection line 3 and control line, nitrocellulose filter after being coated afterwards is put in confining liquid and is closed, it is dried, standby；
2) assemble: on plastic plate, place step 1) the glass fibre prime modulus prepared, then in step 1) place step 1 on the glass fibre element film prepared) nitrocellulose filter prepared, then sample pad is placed in one end of celluloid, the corresponding other end places adsorptive pads, it is dried, i.e. completes.
6. the preparation method of stomach Function detection test kit as claimed in claim 5, it is characterized in that, also including in step A first preparing quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody and the mixture of quantum dot-labeled HP ureB antigen, concrete grammar includes following operating procedure:
A: add modified cyclodextrin in latex particle suspension, mix 24 hours, purification, obtain modified cyclodextrin-latex compound suspension；
B: add quantum dot in modified cyclodextrin-latex compound suspension prepared by step a, mix 24 hours, purification, obtain quantum dot-modified cyclodextrin-latex compound suspension；
C: add activator in quantum dot-modified cyclodextrin-latex compound suspension prepared by step b, activate 15 minutes, purification, add pepsinogen I antibody afterwards, after mixing 2～4 hours, be centrifuged to obtain precipitate particles, by resuspended for precipitate particles confining liquid, room temperature mixes 1 hour, is purified, obtains quantum dot-labeled pepsinogen I antibody；
D: prepare quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB antigen according to the method that above-mentioned steps a～c are same, then quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB antigen are mixed, standby.
7. the preparation method of stomach Function detection test kit as claimed in claim 6, it is characterised in that described modified cyclodextrin is carboxymethyl-modification cyclodextrin or amination modified cyclodextrin.
8. the preparation method of stomach Function detection test kit as claimed in claim 6, it is characterised in that modified cyclodextrin described in step a is 20:1 with the mass ratio of latex particle；Quantum dot described in step b is 10:1 with the mass ratio of cyclodextrin-latex compounds；Pepsinogen I antibody described in step c is 1:20 with the mass ratio of quantum dot-modified cyclodextrin-latex compound.
9. the preparation method of stomach Function detection test kit as claimed in claim 6, it is characterized in that, quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody, the mixing of quantum dot-labeled HP ureB antigen are coated on glass fibre element film by step A method particularly includes: by quantum dot-labeled pepsinogen I antibody, quantum dot-labeled pepsinogen I I antibody and quantum dot-labeled HP ureB antigen mixture even application on glass fibre element film.
10. the preparation method of stomach Function detection test kit as claimed in claim 6, it is characterized in that, pepsinogen I two is resisted by step B, pepsinogen I I bis-resists, HP ureB antigen, sheep anti mouse lgG is coated on the detection line 1 of nitrocellulose filter respectively, detection line 2, on detection line 3 and control line method particularly includes: take concentration respectively and be the pepsinogen I two of 1mg/ml and resist, pepsinogen I I bis-resists, HP ureB antigen and sheep anti-mouse igg, with 1ul/cm, the speed of 10cm/ second sprays to the detection line 1 of cellulose nitrate respectively, detection line 2, on detection line 3 and control line.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107271669A (en) * | 2016-09-23 | 2017-10-20 | 清华大学深圳研究生院 | Propepsin, helicobacter pylori antibody and G17 detection method and its kit |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101680899A (en) * | 2007-05-17 | 2010-03-24 | 明治乳业株式会社 | Method for evaluating gastric mucosal state by quantifying helicobacter pylori antigen in feces |
| CN102520192A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay and kit for quantitative detection of troponin I/creatine kinase isoenzyme/myohemoglobin |
| CN102662059A (en) * | 2012-05-11 | 2012-09-12 | 北京美康生物技术研究中心 | Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof |
| CN104076142A (en) * | 2014-03-05 | 2014-10-01 | 广东医学院附属医院 | Fluorescent microsphere lateral chromatographic detection strip for multiple joint inspection of trace target substances as well as preparation method and application thereof |
| CN104142399A (en) * | 2013-05-08 | 2014-11-12 | 北京美康生物技术研究中心有限责任公司 | Test paper strip using colloidal gold immunochromatographic technology for quantitative determination of serum pepsinogen and preparation method and application thereof |
-
2016
- 2016-05-17 CN CN201610327064.1A patent/CN106018792B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101680899A (en) * | 2007-05-17 | 2010-03-24 | 明治乳业株式会社 | Method for evaluating gastric mucosal state by quantifying helicobacter pylori antigen in feces |
| CN102520192A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay and kit for quantitative detection of troponin I/creatine kinase isoenzyme/myohemoglobin |
| CN102662059A (en) * | 2012-05-11 | 2012-09-12 | 北京美康生物技术研究中心 | Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof |
| CN104142399A (en) * | 2013-05-08 | 2014-11-12 | 北京美康生物技术研究中心有限责任公司 | Test paper strip using colloidal gold immunochromatographic technology for quantitative determination of serum pepsinogen and preparation method and application thereof |
| CN104076142A (en) * | 2014-03-05 | 2014-10-01 | 广东医学院附属医院 | Fluorescent microsphere lateral chromatographic detection strip for multiple joint inspection of trace target substances as well as preparation method and application thereof |
Cited By (16)
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|---|---|---|---|---|
| CN107328938A (en) * | 2016-09-23 | 2017-11-07 | 清华大学深圳研究生院 | Propepsin and helicobacter pylori antibody detection method and its kit |
| CN107271669A (en) * | 2016-09-23 | 2017-10-20 | 清华大学深圳研究生院 | Propepsin, helicobacter pylori antibody and G17 detection method and its kit |
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Denomination of invention: The invention relates to an immunochromatographic kit for gastric function detection and a preparation method thereof Effective date of registration: 20211009 Granted publication date: 20180213 Pledgee: Haidian Beijing science and technology enterprise financing Company limited by guarantee Pledgor: BEIJING MOKOBIO LIFE SCIENCE Co.,Ltd.|NANJING MOKOBIO BIOTECHNOLOGY Co.,Ltd. Registration number: Y2021990000940 |
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Date of cancellation: 20230323 Granted publication date: 20180213 Pledgee: Haidian Beijing science and technology enterprise financing Company limited by guarantee Pledgor: BEIJING MOKOBIO LIFE SCIENCE Co.,Ltd.|NANJING MOKOBIO BIOTECHNOLOGY Co.,Ltd. Registration number: Y2021990000940 |