CN109959796A - A kind of enzyme-linked aptamers absorption detection kit of human serum DKK1 - Google Patents
A kind of enzyme-linked aptamers absorption detection kit of human serum DKK1 Download PDFInfo
- Publication number
- CN109959796A CN109959796A CN201711436359.3A CN201711436359A CN109959796A CN 109959796 A CN109959796 A CN 109959796A CN 201711436359 A CN201711436359 A CN 201711436359A CN 109959796 A CN109959796 A CN 109959796A
- Authority
- CN
- China
- Prior art keywords
- dkk1
- kit
- enzyme
- human serum
- aptamers
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
-
- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57488—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds identifable in body fluids
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to biological product technical field, it is related to a kind of enzyme-linked aptamers absorption detection kit of human serum DKK1 content and preparation method.The kit is made of following component: 96 orifice plates of succinimido modification；Phosphate buffer；Block buffer；TMB developing solution；Detect dilution；Washing buffer；The aptamer of 5 ' amidized identification DKK1 albumen；Goat anti-Human's DKK1 polyclonal antibody of biotin labeling；Standard items: DKK1 albumen；Horseradish peroxidase-labeled Streptavidin；Terminate liquid and sealing plate film.Kit of the invention experiments verify that, show that the kit can detect human serum DKK1 content, as hepatocarcinoma early diagnosis and early warning, method is easy, high specificity, high sensitivity, compared with traditional enzyme-linked immuno sorbent assay kit, detection time is shorter, and accuracy rate is higher, cost is lower, there is preferable application prospect.
Description
Technical field
The invention belongs to field of biotechnology, are related to enzyme-linked aptamers absorption detection kit, and in particular to a kind of people's blood
The clear enzyme-linked aptamers of DKK1 content adsorb detection kit and preparation method.
Background technique
It is dickkopf protein family member prior art discloses DKK1 albumen, which includes two rich in half
The structural domain of cystine, can be specifically in conjunction with one of the receptor of Wnt LRP5/6, to interfere LRP5/6 and Wnt-
The combination of Frizzled compound inhibits the conduction of downstream signal.Another high-affinity receptor of DKK1 is Kremen
(Krm), studies have found that in the presence of Krm2, DKK1 can form ternary complex with LRP6 and Krm2, and should
Endocytosis can rapidly occur for ternary complex；For studies have shown that due to the presence of this endocytosis phenomenon, the LRP6 of film surface can be with
It is removed, to further suppress Wnt/ β-catenin signal.The DKK1 is also proved to increase by restoring β-catenin
OCT4 expression inhibiting Wnt/ β-catenin signal path, this acts in heart, head and forelimb development and plays a key effect.
Studies have shown that DKK1 is seldom expressed in adult body, the only high expression in embryonic tissue and placenta；There is research to demonstrate,prove
Real DKK1 is overexpressed in Hepatocellular Carcinoma, and can not be detected in normal liver tissue；In liver cancer, lung cancer, breast cancer, glue
Detect that secretory DKK1 content improves in the various kinds of cell culture solution such as matter tumor, cervical carcinoma, related research is to 831 clinics
It is found after the DKK1 proficiency assessment of serum sample (424 liver cancer, 98 hepatitis B, 96 cirrhosis, 212 normal controls),
DKK1 reaches 69.% as the detection sensitivity of liver cancer marker, and specificity reaches 90.6%；The liver cancer of AFP negative is suffered from
Person's detection sensitivity is 70.4%, and specificity is 90.0%；Therefore, detecting DKK1 in serum can be used as hepatocarcinoma early diagnosis and pre-
Alert important means and index has important value especially for the liver cancer of AFP negative.
It is single-stranded oligonucleotide of a kind of length less than 100 bases that the prior art, which also discloses aptamers, by itself
Folding can form higher structure, and the identification of specificity is realized with target, and be known as " chemical antibody ".Studies have shown that adaptation
The affinity and specificity of body and target are no less than antibody, and aptamers have that identification target is more various, the R&D cycle more
It is short, performance is more stable, be easier to modification, the more low advantage of cost, have been widely used for the numerous areas such as scientific research, diagnosis, treatment.
Enzyme-linked aptamers absorption detection kit based on aptamers is compared with traditional enzyme-linked immuno sorbent assay kit, when detection
Between it is shorter, accuracy rate is higher, the more low advantage of cost, and present inventor is quasi- to provide a kind of enzyme-linked aptamers absorption detection examination
Agent box, and in particular to a kind of enzyme-linked aptamers absorption detection kit of human serum DKK1 content and preparation method.
Summary of the invention
The object of the present invention is to provide a kind of enzyme-linked aptamers to adsorb detection kit, and in particular to a kind of human serum DKK1
The enzyme-linked aptamers absorption detection kit of content and preparation method.
Detection kit of the invention is the enzyme-linked aptamers absorption qualitative detection kit of human serum DKK1, can be used for standard
Change, DKK1 in stylized ground qualitative detection human serum, be avoided that influence of the differentiation to the accuracy, reliability of detection, is applicable in
High sensitivity is provided in hepatocarcinoma early diagnosis and early warning, the hepatocarcinoma early diagnosis of in particular AFP negative and early warning and special is examined
Disconnected kit.Enzyme-linked aptamers based on aptamers of the invention adsorb detection kit and traditional MBP enzyme linked immuno-adsorbent assay
Kit is compared, and detection time is shorter, and accuracy rate is higher, and cost is lower.
The purpose of the present invention is applied existing by following technical proposals:
A kind of enzyme-linked aptamers adsorptive agent box of human serum DKK1 qualitative detection, which is characterized in that the kit
By as follows at being grouped as:
1) for being coated with 96 orifice plates of the succinimido modification of aptamers；
2) phosphate buffered saline: 137mmol/l (8.0g/l) sodium chloride, 2.7mmol/l (0.2g/l) potassium chloride,
8.1mmol/l (1.15g/l) disodium hydrogen phosphate, 1.5mM (0.2g/l) potassium dihydrogen phosphate, pH 7.4；
3) 1g bovine serum albumin(BSA) Block buffer: is added in 100ml phosphate buffer；
4) TMB developing solution: 0.1mol/l (19.2g/l) citric acid solution 24.3ml adds 0.2mol/l (28.4g/l) phosphoric acid
Disodium hydrogen solution 25.7ml, then plus distilled water 50ml, by 40g o-phenylenediamine dissolution wherein, then plus 30% hydrogen peroxide 0.10ml, use
When Fresh, be protected from light；
5) detect dilution: 1g bovine serum albumin(BSA) is added in 100ml phosphate buffer, and 100 μ g yeast transport ribose core
Acid；
6) washing buffer: 0.5ml Tween-20 is added in 100ml phosphate buffer；
7) aptamer of 5 ' amidized identification DKK1 albumen；
8) Goat anti-Human's DKK1 polyclonal antibody of biotin labeling；
9) standard items: DKK1 albumen；
10) horseradish peroxidase-labeled Streptavidin；
11) terminate liquid: 1mol/l sulfuric acid；
12) sealing plate film.
The enzyme-linked aptamers of human serum DKK1 of the invention adsorb detection kit, are prepared by following methods comprising:
1) using 96 orifice plates (being purchased from Corning Incorporated) of the succinimido modification of coating aptamers；
2) phosphate buffered saline is prepared: by 8.0g sodium chloride, 0.2g potassium chloride, 2.8g ten are added in 1l deionized water
Two hypophosphite monohydrate disodium hydrogens, bis- hypophosphite monohydrate potassium dihydrogen of 0.2g are made, pH value 7.4；
3) it prepares Block buffer: being made by 1g bovine serum albumin(BSA) is added in 100ml phosphate buffer；
4) it prepares TMB developing solution: by 0.1mol/l citric acid solution 24.3ml, adding 0.2mol/l disodium phosphate soln
25.7ml, then plus distilled water 50ml, then by 40g o-phenylenediamine dissolution wherein, then plus 30% hydrogen peroxide 0.10ml be made；
5) it prepares detection dilution: the bovine serum albumin(BSA) and 100 μ g yeast of 1g being added by the phosphate buffer of 100ml
Transfer RNA (tRNA) is made；
6) it prepares washing buffer: being made of the Tween-20 that 0.5ml is added in the phosphate buffer of 100ml；
7) sequence of the aptamer of described 5 ' amidized identification DKK1 albumen are as follows: NH2-5 '-
catatgattaggctgtaacggggctaggcggggatcatt-3'；
8) Goat anti-Human's DKK1 polyclonal antibody of biotin labeling is purchased from R&D Systems company；
9) DKK1 albumen is purchased from the Divine Land Beijing Yi Qiao Science and Technology Ltd.；
10) horseradish peroxidase-labeled Streptavidin is purchased from R&D Systems company；
11) terminate liquid uses 1mol/l sulfuric acid；
12) sealing plate film sealing plate is used.
A kind of human serum DKK1 qualitative detection kit, it is characterised in that the kit also needs to use when in use
To following material:
1) microplate reader (band 450nm)；
2) sample injector, the disposable pipette tips of 10 μ l, 100 μ l, 1000 μ l；
3) blotting paper or Water adsorption towel.
In the present invention, the citric acid, two hypophosphite monohydrate potassium dihydrogens, disodium hydrogen phosphate dodecahydrate, o-phenylenediamine, sulfuric acid
It is that chemistry is pure；The bovine serum albumin(BSA), yeast transfer RNA (tRNA), Tween-20 are that analysis is pure.
Above-mentioned medicament of the present invention can directly be bought from shop.
The chemical component of Tween-20 of the present invention is polyoxyethylene 20 sorbitan monolaurate.
The present invention has carried out testing inspection, wherein depicts examination criteria curve (such as Fig. 1 institute with the kit
Show), to determine detection limit；Testing result shows that kit detection limit can achieve actual sample down to 62.5pg/ml
Testing requirements；Then, using the kit to the DKK1 contents of 30 normal human serums and 30 liver cancer patient blood serums into
Detection is gone, the results show that the DKK1 positive rate in liver cancer is apparently higher than normal group, the p value of detection is 0.0003, there is statistics
Meaning (as shown in Figure 2).
The beneficial effects of the present invention are as follows: the enzyme-linked aptamers of human serum DKK1 of preparation adsorb detection kit, accuracy, can
By property height, it is suitable for detection and DKK1 protein related diseases, including lung cancer, liver cancer, the cancer of the esophagus, cancer of pancreas, myeloma, intrauterine
Film cancer, cervical carcinoma or rheumatoid arthritis；The present invention in particular liver cancer of hepatocarcinoma early diagnosis and early warning and AFP negative is early
Phase diagnosis and early warning provide High sensitivity and special diagnostic kit；Enzyme-linked aptamers based on aptamers of the invention are inhaled
For attached detection kit compared with traditional enzyme-linked immuno sorbent assay kit, detection time is shorter, and accuracy rate is higher, and cost is more
It is low.
Detailed description of the invention
Fig. 1 is the examination criteria curve drawn with kit of the invention.
Fig. 2 is kit detection normal human serum (n=30) and liver cancer patient blood serum (n=30) DKK1 albumen of the present invention
Amount, p=0.0003.
Specific embodiment
Embodiment 1 prepares the enzyme-linked aptamers absorption detection kit of human serum DKK1
1, the detection kit is made of following component:
1) for being coated with 96 orifice plates of the succinimido modification of aptamers；
2) phosphate buffered saline: 137mmol/l (8.0g/l) sodium chloride, 2.7mmol/l (0.2g/l) potassium chloride,
8.1mmol/l (1.15g/l) disodium hydrogen phosphate, 1.5mM (0.2g/l) potassium dihydrogen phosphate, pH 7.4；
3) 1g bovine serum albumin(BSA) Block buffer: is added in 100ml phosphate buffer；
4) TMB developing solution: 0.1mol/l (19.2g/l) citric acid solution 24.3ml adds 0.2mol/l (28.4g/l) phosphoric acid
Disodium hydrogen solution 25.7ml, then plus distilled water 50ml, by 40g o-phenylenediamine dissolution wherein, then plus 30% hydrogen peroxide 0.10ml, use
When Fresh, be protected from light；
5) detect dilution: 1g bovine serum albumin(BSA) is added in 100ml phosphate buffer, and 100 μ g yeast transport ribose core
Acid；
6) washing buffer: 0.5ml Tween-20 is added in 100ml phosphate buffer；
7) aptamer of 5 ' amidized identification DKK1 albumen；
8) Goat anti-Human's DKK1 polyclonal antibody of biotin labeling；
9) standard items: DKK1 albumen；
10) horseradish peroxidase-labeled Streptavidin；
11) terminate liquid: 1mol/l sulfuric acid；
12) sealing plate film.
By following preparation methods:
1) using 96 orifice plates (being purchased from Corning Incorporated) of the succinimido modification of coating aptamers；
2) phosphate buffered saline is prepared: by 8.0g sodium chloride, 0.2g potassium chloride, 2.8g ten are added in 1l deionized water
Two hypophosphite monohydrate disodium hydrogens, bis- hypophosphite monohydrate potassium dihydrogen of 0.2g are made, pH value 7.4；
3) it prepares Block buffer: being made by 1g bovine serum albumin(BSA) is added in 100ml phosphate buffer；
4) it prepares TMB developing solution: by 0.1mol/l citric acid solution 24.3ml, adding 0.2mol/l disodium phosphate soln
25.7ml, then plus distilled water 50ml, then by 40g o-phenylenediamine dissolution wherein, then plus 30% hydrogen peroxide 0.10ml be made；
5) it prepares detection dilution: the bovine serum albumin(BSA) and 100 μ g yeast of 1g being added by the phosphate buffer of 100ml
Transfer RNA (tRNA) is made；
6) it prepares washing buffer: being made of the Tween-20 that 0.5ml is added in the phosphate buffer of 100ml；
7) sequence of the aptamer of described 5 ' amidized identification DKK1 albumen are as follows: NH2-5 '-
catatgattaggctgtaacggggctaggcggggatcatt-3'；
8) Goat anti-Human's DKK1 polyclonal antibody of biotin labeling is purchased from R&D Systems company；
9) DKK1 albumen is purchased from the Divine Land Beijing Yi Qiao Science and Technology Ltd.；
10) horseradish peroxidase-labeled Streptavidin is purchased from R&D Systems company；
11) terminate liquid uses 1mol/l sulfuric acid；
12) sealing plate film sealing plate is used.
The citric acid, two hypophosphite monohydrate potassium dihydrogens, disodium hydrogen phosphate dodecahydrate, o-phenylenediamine, sulfuric acid are chemistry
It is pure；The bovine serum albumin(BSA), yeast transfer RNA (tRNA), Tween-20 are that analysis is pure；
The above-mentioned medicament can directly be bought from shop.
2, prepare following material:
1) microplate reader (band 450nm)；
2) sample injector, the disposable pipette tips of 10 μ l, 100 μ l, 1000 μ l；
3) blotting paper or Water adsorption towel.
3, human serum sample prepares and stores: taking a blood sample on an empty stomach, natural storage is centrifuged after 1-2 hours.Separation gel is wherein housed
The direct 3000rmp of the blood sample adopted of pipe be centrifuged 15 minutes；The blood that simple glass or plastic tube are adopted then placed 37 degree after 30 minutes
3000 revs/min are centrifuged 10 minutes, and treated, and blood sample to be checked is closed is stored in 2-8 DEG C of refrigerator；
4, it is detected by following detection program:
1) be coated with: amber is added with the detection amidized DKK1 aptamers of diluted 5 ' to 50nmol/l in the detection same day
In 96 orifice plates of amber imide modification, every 100 μ l of hole, with being stored at room temperature 30 minutes after sealing plate film sealing plate；
2) liquid in enzyme linked plate holes is sucked out, with washing buffer concussion washing totally 3 times, pats dry rear centrifuge dripping every time；
3) close: 300ul Block buffer is added in every hole, with being stored at room temperature 1 hour after sealing plate film sealing plate；
4) it is loaded: setting blank well, gauge orifice, sample to be tested hole, 100 holes μ l/ respectively.DKK1 albumen as standard items, with
Phosphate buffer gradient dilution (1:100；1:1000；1:2000；1:4000；1:8000) with being stored at room temperature after sealing plate film sealing plate
1 hour.
5) liquid in enzyme linked plate holes is poured out, with washing buffer concussion washing 3 times, pats dry rear centrifuge dripping every time；
6) the Goat anti-Human DKK1 polyclonal antibody of 50ng/ml biotin labeling is added in every hole (with detection diluted)
100 μ l, with being stored at room temperature 1 hour after sealing plate film sealing plate；
7) liquid in enzyme linked plate holes is poured out, with washing buffer concussion washing 3 times, pats dry rear centrifuge dripping every time；
8) horseradish peroxidase-labeled Streptavidin (with detection dilution 1:200 dilution) 100 μ l are added in every hole, use
It is stored at room temperature after sealing plate film sealing plate 30 minutes；
9) liquid in enzyme linked plate holes is poured out, with washing buffer concussion washing 3 times, pats dry rear centrifuge dripping every time；
10) develop the color: TMB developing solution (ready-to-use) 100 μ l is added in every hole, with 37 DEG C of waters bath with thermostatic control 20 after sealing plate film sealing plate
Minute, 100 hole μ l/ of terminate liquid is added dropwise and terminates reaction.
11) ELISA Plate is placed in microplate reader, is returned to zero with blank control wells, wavelength 450nm reads result.
Preliminary identification has been carried out in the present embodiment, depicts examination criteria curve (as shown in Figure 1) with the kit,
To determine detection limit；The results show that the kit detection limit can achieve the detection of actual sample down to 62.5pg/ml
It is required that；Then, it is detected with DKK1 content of the kit to 30 normal human serums and 30 liver cancer patient blood serums, as a result
It shows, the DKK1 positive rate in liver cancer is apparently higher than normal group, and the p value of detection is 0.0003, statistically significant (such as Fig. 2 institute
Show).
Claims (4)
1. a kind of enzyme-linked aptamers of human serum DKK1 adsorb detection kit, which is characterized in that the kit includes:
1) for being coated with 96 orifice plates of the succinimido modification of aptamers；
2) phosphate buffered saline；
3) Block buffer；
4) TMB developing solution；
5) dilution is detected；
6) washing buffer；
7) aptamer of 5 ' amidized identification DKK1 albumen；
8) Goat anti-Human's DKK1 polyclonal antibody of biotin labeling；
9) standard items；
10) horseradish peroxidase-labeled Streptavidin；
11) terminate liquid；With,
12) sealing plate film.
2. the enzyme-linked aptamers of human serum DKK1 according to claim 1 adsorb detection kit, which is characterized in that described
Identify the aptamer of DKK1 albumen, sequence is
5’-catatgattaggctgtaacggggctaggcggggatcatt-3’。
3. the enzyme-linked aptamers of human serum DKK1 according to claim 1 adsorb detection kit, which is characterized in that described
Kit using when also need material:
1) microplate reader, band 450nm；
2) sample injector, the disposable pipette tips of 10 μ l, 100 μ l, 1000 μ l；
3) blotting paper or Water adsorption towel.
4. the enzyme-linked aptamers of human serum DKK1 according to claim 1 adsorb detection kit, which is characterized in that with DKK1
Protein related diseases include lung cancer, liver cancer, the cancer of the esophagus, cancer of pancreas, myeloma, carcinoma of endometrium, cervical carcinoma or rheumatoid arthrosis
It is scorching.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711436359.3A CN109959796B (en) | 2017-12-26 | 2017-12-26 | Human serum DKK1 enzyme-linked aptamer adsorption detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711436359.3A CN109959796B (en) | 2017-12-26 | 2017-12-26 | Human serum DKK1 enzyme-linked aptamer adsorption detection kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109959796A true CN109959796A (en) | 2019-07-02 |
| CN109959796B CN109959796B (en) | 2022-05-10 |
Family
ID=67022518
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711436359.3A Active CN109959796B (en) | 2017-12-26 | 2017-12-26 | Human serum DKK1 enzyme-linked aptamer adsorption detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109959796B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112748247A (en) * | 2020-12-23 | 2021-05-04 | 复旦大学附属华山医院 | Application of antibody binding agent in preparation of auxiliary detection product for vitiligo stage and detection kit for vitiligo stage identification |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102081092A (en) * | 2009-11-30 | 2011-06-01 | 武汉大学 | Kit and detection method for mycobacterium tuberculosis |
| CN102507951A (en) * | 2011-11-25 | 2012-06-20 | 广东药学院 | Enzyme-linked immuno sorbent assay (ELISA) kit for performing joint detection on tumor marker |
| US20120201810A1 (en) * | 2011-02-01 | 2012-08-09 | Ng Liu Irene Oi Lin | Use of Anti-DKK-1 Monoclonal Antibodies for the Treatment of Liver Cancer |
| CN103487580A (en) * | 2012-06-08 | 2014-01-01 | 上海市肿瘤研究所 | Application of DKK1 as diagnostic marker |
| CN104830867A (en) * | 2015-06-07 | 2015-08-12 | 杨洋 | Aptamer capable of being specifically combined with DKK1 protein in cancer cells |
| CN105203760A (en) * | 2015-07-24 | 2015-12-30 | 中国人民解放军第二军医大学 | PSMD4 protein ELISA detection kit as well as detection method and application thereof |
| CN105588941A (en) * | 2015-02-28 | 2016-05-18 | 苏州飞康生物医药有限公司 | Enzyme-linked immunosorbent assay kit for detecting concentration of tumor marker DKK1 |
-
2017
- 2017-12-26 CN CN201711436359.3A patent/CN109959796B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102081092A (en) * | 2009-11-30 | 2011-06-01 | 武汉大学 | Kit and detection method for mycobacterium tuberculosis |
| US20120201810A1 (en) * | 2011-02-01 | 2012-08-09 | Ng Liu Irene Oi Lin | Use of Anti-DKK-1 Monoclonal Antibodies for the Treatment of Liver Cancer |
| CN102507951A (en) * | 2011-11-25 | 2012-06-20 | 广东药学院 | Enzyme-linked immuno sorbent assay (ELISA) kit for performing joint detection on tumor marker |
| CN103487580A (en) * | 2012-06-08 | 2014-01-01 | 上海市肿瘤研究所 | Application of DKK1 as diagnostic marker |
| CN105588941A (en) * | 2015-02-28 | 2016-05-18 | 苏州飞康生物医药有限公司 | Enzyme-linked immunosorbent assay kit for detecting concentration of tumor marker DKK1 |
| CN104830867A (en) * | 2015-06-07 | 2015-08-12 | 杨洋 | Aptamer capable of being specifically combined with DKK1 protein in cancer cells |
| CN105203760A (en) * | 2015-07-24 | 2015-12-30 | 中国人民解放军第二军医大学 | PSMD4 protein ELISA detection kit as well as detection method and application thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112748247A (en) * | 2020-12-23 | 2021-05-04 | 复旦大学附属华山医院 | Application of antibody binding agent in preparation of auxiliary detection product for vitiligo stage and detection kit for vitiligo stage identification |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109959796B (en) | 2022-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107817354A (en) | A kind of chemiluminescence detection kit of interleukin 6 and preparation method thereof | |
| CN101858914B (en) | Reagent strip for testing syphilis specific total antibodies through gold immunochromatographic assay and preparation method thereof | |
| CN102043058A (en) | Detection kit of acetyl amantadine for predicting tumors | |
| CN109100516A (en) | A kind of detection cytomegalovirus IgG antibody chemical luminescence immune assay determination reagent kit and preparation method thereof | |
| CN104198716A (en) | Animal brucella antibody gold mark rapid detection kit | |
| CN101592661B (en) | Brucellosis antibody competitive enzyme-linked immunosorbent assay reagent kit | |
| CN102368071B (en) | Chemiluminescent immunoassay kit for detecting mycoplasma pneumoniae IgM antibody | |
| CN105842464B (en) | Joint based on up-converting phosphor technology quantitatively detects uNGAL and uCr device and preparation method thereof | |
| CN101581726A (en) | New-generation brucellosis antibody competition enzyme-linked immunosorbent adsorption test detection kit | |
| JP4976068B2 (en) | Simple immunoassay specimen suspension and assay method | |
| EP3349010B1 (en) | Anti-vla-4 related assays | |
| PT88615B (en) | EQUIPMENT AND METHOD OF IMMUNOMETRIC DOSAGE APPLICABLE TO COMPLETE CELLS | |
| CN109959796A (en) | A kind of enzyme-linked aptamers absorption detection kit of human serum DKK1 | |
| CN102368068B (en) | Kit for detecting chlamydia pneumoniae IgM antibody | |
| CN104730231B (en) | A kind of sample buffer for fluorescence immunoassay detection by quantitative and application thereof | |
| CN103389374A (en) | Colloid gold kit for detecting mycoplasma pneumoniae by micro whole-blood loading method | |
| CN106290863A (en) | A kind of human hepatitis C virus (HCV) saliva/urine antibody colloidal gold detection kit and preparation method thereof | |
| CN103288964B (en) | Albendazole-2-amino sulfone residue detection monoclonal antibody as well as preparation method and application thereof | |
| CN103954763A (en) | Method for detecting capability of in-vitro promotion of phosphorylation modification of disease-related protein in body fluid of subject | |
| Okumura et al. | Detection of white spot syndrome virus (WSSV) from hemolymph of Penaeid shrimps Penaeus japonicus by reverse passive latex agglutination assay using high-density latex particles | |
| RU2234704C2 (en) | Antibody analysis | |
| JP5911404B2 (en) | Simple membrane assay and kit | |
| JP5456056B2 (en) | Detection of IFI16 in body fluids | |
| CN111596069A (en) | Application and product of HSP10 in early warning, diagnosis and prognosis evaluation of POP | |
| CN103698533A (en) | Use of Apo-Al protein in preparation of serum marker for early screening or diagnosis of lung cancer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |