CN103604918A - Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate - Google Patents
Luminescent substrate, use of luminescent substrate and detection kit containing luminescent substrate Download PDFInfo
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- CN103604918A CN103604918A CN201310625446.9A CN201310625446A CN103604918A CN 103604918 A CN103604918 A CN 103604918A CN 201310625446 A CN201310625446 A CN 201310625446A CN 103604918 A CN103604918 A CN 103604918A
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- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
- G01N21/763—Bioluminescence
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- 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
- G01N33/57423—Specifically defined cancers of lung
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- 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
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
Abstract
The invention discloses a luminescent substrate, a use of the luminescent substrate and a detection kit containing the luminescent substrate. The luminescent substrate has high sensitivity and high stability. The luminescent substrate comprises a solution A and a solution B. The solution A comprises 0.5-8mmol/L of luminal, 0.5-10mmol/L of p-lodophenol, and 0.1mol/L of a Tris-HCl buffer solution. The solution B comprises 1-10mmol/L of urea peroxide and 0.1mol/L of the Tris-HCl buffer solution. The kit comprises a monoclonal antibody-coated microporous plate, HRP-labeled monoclonal antibodies, a freeze-dried standard, a sample diluent, a concentrated washing liquid and the chemiluminescent substrate.
Description
Technical field
The invention discloses a kind of luminous substrate, the invention also discloses the application of this luminous substrate and the detection kit that contains this luminous substrate.
Background technology
The research of labelled immune analytical technology and application development are rapid nearly ten years, have been widely used in each field of biomedical fundamental research and clinical disease diagnosis.For detection of the method for serological index, mainly comprise radioactive isotope immunoassay, enzyme linked immunosorbent assay and the chemistry immunoassay of giving out light.
At present, the quantitative immunologic detection method that clinical field is most widely used is mainly chemiluminescence immune assay (CLIA), it is a kind of responsive skeptophylaxis determination method, have the advantages such as radio-immunity (RIA) high sensitivity and enzyme linked immunological (ELISA) be easy to operate concurrently, overcome again radio-immunity and enzyme linked immunological shortcoming separately, be one of optimal method of clinical immune detection simultaneously.
Chemiluminescence immunoassay can be divided into three major types according to the difference of label, i.e. chemiluminescence immune assay, chemiluminescence enzyme immunoassay and electrochemiluminescence immunoassay.Wherein with chemiluminescence enzyme immunoassay (the Chemiluminescence Enzyme Immol/Lunoassay of horseradish peroxidase (HRP) mark, CLEIA) be to carry out immune response with HRP mark bioactivator (as the antigen of enzyme labeling or antibody), enzyme on immune response compound remakes for luminous substrate, excite chemical luminous substrate to produce light signal, utilize luminous signal analyzer to carry out luminescence assays.HRP has many luminous substrate, but the most frequently used luminous substrate is luminol and derivant thereof.In CLEIA, use HRP enzyme labelled antibody to carry out after immune response, under alkali condition, HRP and oxygenant (H2O2) act on luminous substrate luminol makes it that chemical reaction and then luminous occur, and utilizes light signal analyzer can measure this light signal.But this traditional chemical luminous system (HRP-H2O2-luminol) exists luminous strong ℃ low, is difficult for the shortcomings such as measurement.Afterwards, research finds to add enhancing luminous agent in luminescent system, as the bromine acidification compound that contains substituent phenolic compound (as 4-(3-thienyl) phenolic compound), naphthols, replacement, Acetanilide Derivatives etc., can greatly strengthen luminous signal, and within the longer time, keep stable, be convenient to duplicate measurements, thereby improve sensitivity for analysis and accuracy.
Primary carcinoma of liver, especially hepatocellular carcinoma (Hepatocellular Carcinoma), as one of modal malignant tumour in the world, also be one of poor malignant tumour of prognosis effect in various entity tumors in the world, and thering is the disease feature of maintaining close ties with hepatitis B, cirrhosis etc., its morbidity rate is ascendant trend year by year in China especially in recent years.Therefore, the early warning technical research of liver cancer has very important researching value and social effect for the incidence trend of prevent liver cancer, predicting liver cancer.The current diagnosing cancer of liver mark AFP of widespread use clinically, still, for the diagnosis of early liver cancer, its susceptibility, specificity are all undesirable.Therefore wish clinically to obtain than alpha-fetoprotein specificity and the higher new serological index of sensitivity, or can carry out complementary index with AFP.Wherein GP73 (GolgiProtein73, GP73) GP73 be research in recent years find most possibly become the especially blood serum designated object of early liver cancer diagnosis of better diagnosing cancer of liver.For early liver cancer (T1: single tubercle, diameter <2cm and T2: single tubercle, diameter is between 2cm～5cm or be less than three tubercles, each diameter <3cm) diagnosis, the susceptibility of GP73 is significantly better than AFP, and in the HCC patient of AFP level lower than 20 μ g/L, nearly 57% crowd GP73 level significantly raises, this shows that HCC patient GP73 level significantly raises, and therefore, for the diagnosis of early liver cancer, GP73 is better than AFP.
Summary of the invention
For the problems referred to above, the object of the present invention is to provide the chemical luminous substrate of a kind of high sensitivity, high stability, and the kit that the purposes of this luminous substrate is provided and has contained this luminous substrate, this kit is easy to use, highly sensitive.
For solving the problems of the technologies described above, last technical scheme provided by the invention is such: this luminous substrate is comprised of A, B liquid, and described A liquid comprises following component: 0.5～8mmol/L luminol, 0.5～10mmol/L are to iodophenol, 0.1mol/L Tris-HCl damping fluid; Described B liquid comprises following component: 1～10mmol/L urea peroxide, 0.1mol/L Tris-HCl damping fluid.
More excellent, above-mentioned luminous substrate, described A liquid comprises following component: 2.5mmol/L luminol, 3.0mmol/L are to iodophenol, 0.1mol/L Tris-HCl; Described B liquid comprises following component: 1.3mmol/L peroxidating, 0.1mol/L Tris-HCl.
Further, above-mentioned luminous substrate, the pH value of described Tris-HCll damping fluid is 8.5.
Further, above-mentioned luminous substrate, described Tris-HCl damping fluid comprises the component of following mass percent: 0.05～0.3%BSA, 0.01～0.05%Tween-20,0.01～0.1%proclin300, with HCl, regulates pH to 8.5.
Another one technical scheme of the present invention is that this luminous substrate is as the application of preparation ELISA immue quantitative detection reagent box.
Last technical scheme of the present invention is the kit that contains this luminous substrate, and described kit comprises and is coated with the microwell plate of monoclonal antibody, the monoclonal antibody of HRP mark, freeze-drying standard items, sample diluting liquid, concentrated cleaning solution and chemical luminous substrate.
Further, above-mentioned kit, described monoclonal antibody is GP73 monoclonal antibody specific 5F10; The monoclonal antibody of described HRP mark is GP73 monoclonal antibody 5B12*HRP.
11, further, above-mentioned kit, the 0.01mol/L Tris-HCl damping fluid containing 1%BSA, 5% hyclone, 0.1%Proclin300 that described sample diluting liquid is pH7.4; Described concentrated cleaning solution is the PBST concentrated cleaning solution containing 0.05%Tween-20.
Compare with existing prior art, technical scheme tool provided by the invention has the following advantages:
1, luminous substrate provided by the invention, light signal is strong, highly sensitive, good stability, and cost is low;
2, kit provided by the invention, utilize this substrate to carry out chemiluminescence ELISA and quantitatively detect GP73 protein content, the sensitivity and the specificity that detect GP73 have greatly been improved, can be used for monitoring the development degree of early diagnosis, cirrhosis and the chronic hepatitis of liver cancer, for prevention, diagnosis and the treatment of liver cancer provides stronger support.
Accompanying drawing explanation
Fig. 1 is GP73 chemical luminescent analysis reagent kid Code's Sensitivity curve map;
Fig. 2 is the chemical luminous substrate and similar chemical luminous substrate comparative analysis schematic diagram on market in embodiment 1;
The clinical sample research schematic diagram of Fig. 3 kit provided by the invention.
Embodiment
Below in conjunction with embodiment; claim of the present invention is described in further detail; but do not form any limitation of the invention, the modification of the limited number of time that anyone makes within the scope of the claims in the present invention, still in claim protection domain of the present invention.
The preparation of embodiment 1 Chemoluminescent substrate
Substrate formula is as follows:
Formula 1:
Luminous agent solution A liquid: luminol 2.5mmol/L, the Tris-HCl damping fluid to iodophenol 3.0mmol/L, pH8.5;
Oxidizing agent solution B liquid: the Tris-HCl damping fluid of urea peroxide 1.3mmol/L, pH8.5;
Formula 2:
Luminous agent solution A liquid: luminol 0.5mmol/L, the Tris-HCl damping fluid to iodophenol 5.0mmol/L, pH8.5;
Oxidizing agent solution B liquid: the Tris-HCl damping fluid of urea peroxide 1mmol/L, pH8.5;
Formula 3:
Luminous agent solution A liquid: luminol 8mmol/L, the Tris-HCl damping fluid to iodophenol 10mmol/L, pH8.5;
Oxidizing agent solution B liquid: the Tris-HCl damping fluid of urea peroxide 10mmol/L, pH8.5;
Formula 4:
Luminous agent solution A liquid: luminol 5mmol/L, the Tris-HCl damping fluid to iodophenol 0.3mmol/L, pH8.5;
Oxidizing agent solution B liquid: the Tris-HCl damping fluid of urea peroxide 5mmol/L, pH8.5;
In formula 1, the collocation method of Tris-HCl damping fluid, A liquid and B liquid is:
The preparation of damping fluid: get Tris12.1g, bovine serum albumin(BSA) 1.0g, Tween-200.5ml, Proclin-3001.0ml, dissolves with 800ml distilled water, with the pH to 8.5 of concentrated hydrochloric acid regulator solution, with distilled water, be settled to 1000ml, be 0.1mol/LTris-HCl damping fluid, contain 0.1%BSA, 0.05%Tween-20,0.05%Proclin300, pH8.5.
The preparation of luminous agent solution A liquid: take respectively luminol 0.4429g, to iodophenol 0.6600g, wherein luminol is first used the dissolution of sodium hydroxide of 0.1M, dimethyl formamide for iodophenol (DMF) is dissolved, then above-mentioned two kinds of solution are settled to 1000ml with Tris-HCl damping fluid, 0.22um membrane filtration, 4 ℃ keep in Dark Place.
The preparation of oxidizing agent solution B liquid: take urea peroxide 0.1200g, with Tris-HCl damping fluid, dissolve, and be settled to 1000ml, 0.22um membrane filtration, 4 ℃ keep in Dark Place.
The preparation method of Tris-HCl damping fluid, A liquid and B liquid in formula 2 to 4 is similar with Tris-HCl damping fluid, A liquid and B liquid and preparation method thereof in formula 1, only needs according to the consumption of concrete each component of amount concentration adjustment.
Kit provided by the invention comprises and is coated with the microwell plate of monoclonal antibody, the chemical luminous substrate of any one formula in the monoclonal antibody of HRP mark, freeze-drying standard items, sample diluting liquid, concentrated cleaning solution and embodiment 1.
GP73 chemical luminescent analysis reagent kid take below as example, there is the preparation method of this detection kit of explanation, also can be applied to equally the detection of other albumen.
1, the preparation of solid-phase coating plate
A) coated: GP73 monoclonal antibody 5F10 is with the coated microwell plate of 2ug/ml concentration, 100ul/ hole, 4 ℃ of coated 18h;
B) sealing: wash plate 2 times, pat dry, add confining liquid 200ul/ hole, 37 ℃ of 1.5h.
C) envelope: abandon confining liquid, pat dry, be dried 1h at 37 ℃ of incubators, carry out immediately vacuum sealing bag.
2, the preparation of standard items: the GP73 albumen of eukaryotic expression purifying, by sample diluting liquid (being specially which kind of material) doubling dilution to concentration, be 31.25ng/ml, 15.6ng/ml, 7.8ng/ml, 3.9ng/ml, 1.95ng/ml, 0.98ng/ml, 0ng/ml, through 0.22um membrane filtration, the standard items packing 500ul/ pipe of every kind of concentration, obtains freeze-drying standard items by freeze drying.
3, the preparation of enzyme labelled antibody: the sodium periodate method-sodium borohydride method mark horseradish peroxidase HRP of GP73 monoclonal antibody 5B12 through improveing, the i.e. NaIO through 0.1mol/L at the lower HRP of low pH (pH4.4)
4oxidation forms hydroformylation enzyme, and this Fu Shi alkali that the amino of hydroformylation enzyme and antibody molecule 5B12 is connected to form can further be used the NaBH of 4mg/ml
4reduction obtains stable enzymic-labelled antibody.The 5B12*HRP obtaining with antibody diluent be the 0.01mol/L Tris-HCl damping fluid containing 1%BSA, 5% hyclone, 0.1%Proclin300 of pH7.4 by 1/5000 dilution, through 0.22um membrane filtration, packing 12ml/ pipe.
4, preparation concentrated cleaning solution: prepare 20 * the PBST concentrated cleaning solution containing 0.05%Tween-20.
5, preparation Chemoluminescent substrate: prepare respectively luminous agent solution A liquid and oxidizing agent solution B liquid, through 0.22um membrane filtration, rear packing 6ml/ pipe, keeps in Dark Place.
6, be assembled into finished product kit, be placed in 2-8 ℃ of preservation.
The using method of embodiment 3 kits of the present invention
The concrete operations of the GP73 chemical luminescent analysis reagent kid of preparing with embodiment 2 are as follows:
1, reagent, sample preparation
A) kit is from 4 ℃ of taking-ups, and equilibrium at room temperature is placed 20～30min;
B) the every pipe of freeze-drying standard items adds 500ul pure water fully to dissolve;
C) concentrated cleaning solution with pure water 1/20 being diluted to 1 * cleansing solution.
D) before use, sample to be tested is placed in to equilibrium at room temperature 20-30min, shakes and mix gently, if sediment appears in sample, after should centrifugally removing precipitation, use, detect the sensing range that sample used should meet kit, if excessive concentration, after available sample diluting liquid dilution, row detects again.
2, operation steps
A) take out coated plate, add respectively standard items and testing sample, 100ul/ hole, hatches 50min for 37 ℃;
B) coated plate pats dry, and adds ELIAS secondary antibody 5B12*HRP, and 100ul/ hole, hatches 30min for 37 ℃;
C) wash plate 5 times, the chemical luminous substrate that adds equal-volume to mix to prepare, 100ul/ hole, carries out the Luminescence value of reading and detects in 1min;
D) with luminous value and the respective concentration drawing standard curve of each standard items, draw the relation equation formula of luminous value and concentration, the luminous value substitution formula of sample to be measured is calculated to GP73 protein content.
The Code's Sensitivity of embodiment 4GP73 chemical luminescent analysis reagent kid
The kit that uses embodiment 2 preparations, detects by step described in embodiment 3, and its result is as shown in table 1 and Fig. 1.The typical curve linear relationship that can find out kit drafting of the present invention is good, and sensing range is at 0.98～31.25ng/ml.
Table 1
Embodiment 5 chemical luminous substrate of the present invention and similar chemical luminous substrate comparative analysis on market
The supersignal ELSA Femto Maximum sensitivity subatract of substrate solution described in embodiment 1 and Thermo is contrasted.Press embodiment 2 and prepare GP73 chemical luminescent analysis reagent kid, respectively by the RLU value of two substrate detection kit Plays product and Quality Control positive and negative sample, comparative analysis chemical luminous substrate of the present invention and import substrate detect respectively standard items RLU and signal to noise ratio (S/N ratio), its result as shown in Table 2 and Figure 3, can see: compare with import substrate, although the RLU value of substrate of the present invention is lower, but its signal to noise ratio (S/N ratio) P/N value is higher than import substrate, and the cost of substrate of the present invention is far below import substrate.
Table 2
The clinical sample research of embodiment 6 kits of the present invention
By embodiment 2 kit of assembling, a collection of clinical blood serum sample is comprised to 40 parts of severe viral hepatitis, 39 parts of liver cirrhosis patients, 50 parts, 66 parts of liver cancer patients and normal human serum sample, detect contained GP73 protein content in serum, with Graphpad Prism5, carry out statistical analysis, its result as shown in Figure 3, can find out that hepatopathy patient serum GP73 content is far above normal human serum GP73, wherein in heavy type hepatitis serum, GP73 content is the highest, and in liver cancer patient blood serum, GP73 content is higher than GP73 content in serum of cirrhosis patients but there was no significant difference.
Claims (10)
1. a luminous substrate, is characterized in that, A, B liquid, consists of, and described A liquid comprises following component: 0.5～8mmol/L luminol, 0.5～10mmol/L are to iodophenol, 0.1mol/L Tris-HCl damping fluid; Described B liquid comprises following component: 1～10mmol/L urea peroxide, 0.1mol/L Tris-HCl damping fluid.
2. luminous substrate according to claim 1, is characterized in that, described A liquid comprises following component: 2.5mmol/L luminol, 3.0mmol/L are to iodophenol, 0.1mol/L Tris-HCl; Described B liquid comprises following component: 1.3mmol/L urea peroxide, 0.1mol/L Tris-HCl.
3. luminous substrate according to claim 1 and 2, is characterized in that, the pH value of described Tris-HCl is 8.5.
4. luminous substrate according to claim 1 and 2, it is characterized in that, described Tris-HCl damping fluid comprises the component of following mass percent: 0.05～0.3%BSA, 0.01～0.05%Tween-20,0.01～0.1%proclin300, with HCl, regulates pH to 8.5.
5. luminous substrate claimed in claim 1 is as the application of preparation ELISA immue quantitative detection reagent box.
6. the kit containing the luminous substrate described in claim 1, it is characterized in that, described kit comprises and is coated with the microwell plate of monoclonal antibody, the monoclonal antibody of HRP mark, freeze-drying standard items, sample diluting liquid, concentrated cleaning solution and chemical luminous substrate.
7. kit according to claim 6, is characterized in that, described monoclonal antibody is GP73 monoclonal antibody specific 5F10.
8. kit according to claim 6, is characterized in that, the monoclonal antibody of described HRP mark is GP73 monoclonal antibody 5B12*HRP.
9. kit according to claim 6, is characterized in that, the 0.01mol/L Tris-HCl damping fluid containing 1%BSA, 5% hyclone, 0.1%Proclin300 that described sample diluting liquid is pH7.4.
10. kit according to claim 6, is characterized in that, described concentrated cleaning solution is the PBST concentrated cleaning solution containing 0.05%Tween-20.
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| CN104020162A (en) * | 2014-06-27 | 2014-09-03 | 中国农业科学院农业质量标准与检测技术研究所 | Chemiluminiscence sensitization liquid and preparation method thereof |
| CN104049080A (en) * | 2014-06-27 | 2014-09-17 | 中国农业科学院农业质量标准与检测技术研究所 | Chemiluminescence sensibilization liquid and preparation method thereof |
| CN104237520A (en) * | 2014-09-30 | 2014-12-24 | 博奥赛斯（天津）生物科技有限公司 | Hepatitis c virus antigen-antibody joint detection reagent box and preparation method thereof |
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Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06169797A (en) * | 1992-10-09 | 1994-06-21 | Meidensha Corp | Method for measuring hrp |
| US20010046688A1 (en) * | 2000-01-28 | 2001-11-29 | Giri Brij Pal | Novel stabilized formulations for chemiluminescent assays |
| CN1661370A (en) * | 2004-02-23 | 2005-08-31 | 西安君达生物科技有限公司 | High stable method of chemiluminescence accelerated by enzyme and substrate |
| CN101086500A (en) * | 2006-06-09 | 2007-12-12 | 杨致亭 | Mid-pregnancy down's syndrome prenatal screening reagent kit |
| WO2008060394A2 (en) * | 2006-10-27 | 2008-05-22 | Inova Diagnostics, Inc. | Methods and assays for detecting gp73-specific autoantibodies |
| CN101571483A (en) * | 2008-04-29 | 2009-11-04 | 北京科美东雅生物技术有限公司 | Chemoluminescent substrate |
| CN101988926A (en) * | 2009-08-03 | 2011-03-23 | 中国科学院广州生物医药与健康研究院 | Sandwich ELISA quantitative detection method of Golgi protein GP73 and detection kit thereof |
| CN102156121A (en) * | 2010-12-31 | 2011-08-17 | 中生北控生物科技股份有限公司 | Chemiluminescent substrate liquid applied to detection system taking horseradish peroxidase (HRP) as enzymatic reaction |
| CN102367270A (en) * | 2011-06-30 | 2012-03-07 | 同昕生物技术（北京）有限公司 | Preparation method of cyclosporin A haptin and enzymelinked immunosorbent quantitative detection kit of cyclosporin A |
| CN102435598A (en) * | 2011-09-06 | 2012-05-02 | 山东博科生物产业有限公司 | Stable HRP enzymatic enhanced chemiluminescent substrate solution |
| CN102445540A (en) * | 2011-10-11 | 2012-05-09 | 山东莱博生物科技有限公司 | Kit for quantitatively detecting HCV-aAg concentration in human serum by virtue of polystyrene microspheres |
| CN102749456A (en) * | 2012-06-26 | 2012-10-24 | 博奥赛斯（天津）生物科技有限公司 | Kit for chemilumineseent quantitative immunoassay of angiotensin I and preparation method thereof |
| CN102798718A (en) * | 2012-08-31 | 2012-11-28 | 福建省洪诚生物药业有限公司 | Previous S1 antigen detection kit for hepatitis B virus and preparation method of previous S1 antigen detection kit |
-
2013
- 2013-11-28 CN CN201310625446.9A patent/CN103604918A/en active Pending
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06169797A (en) * | 1992-10-09 | 1994-06-21 | Meidensha Corp | Method for measuring hrp |
| US20010046688A1 (en) * | 2000-01-28 | 2001-11-29 | Giri Brij Pal | Novel stabilized formulations for chemiluminescent assays |
| CN1661370A (en) * | 2004-02-23 | 2005-08-31 | 西安君达生物科技有限公司 | High stable method of chemiluminescence accelerated by enzyme and substrate |
| CN101086500A (en) * | 2006-06-09 | 2007-12-12 | 杨致亭 | Mid-pregnancy down's syndrome prenatal screening reagent kit |
| WO2008060394A2 (en) * | 2006-10-27 | 2008-05-22 | Inova Diagnostics, Inc. | Methods and assays for detecting gp73-specific autoantibodies |
| CN101571483A (en) * | 2008-04-29 | 2009-11-04 | 北京科美东雅生物技术有限公司 | Chemoluminescent substrate |
| CN101988926A (en) * | 2009-08-03 | 2011-03-23 | 中国科学院广州生物医药与健康研究院 | Sandwich ELISA quantitative detection method of Golgi protein GP73 and detection kit thereof |
| CN102156121A (en) * | 2010-12-31 | 2011-08-17 | 中生北控生物科技股份有限公司 | Chemiluminescent substrate liquid applied to detection system taking horseradish peroxidase (HRP) as enzymatic reaction |
| CN102367270A (en) * | 2011-06-30 | 2012-03-07 | 同昕生物技术（北京）有限公司 | Preparation method of cyclosporin A haptin and enzymelinked immunosorbent quantitative detection kit of cyclosporin A |
| CN102435598A (en) * | 2011-09-06 | 2012-05-02 | 山东博科生物产业有限公司 | Stable HRP enzymatic enhanced chemiluminescent substrate solution |
| CN102445540A (en) * | 2011-10-11 | 2012-05-09 | 山东莱博生物科技有限公司 | Kit for quantitatively detecting HCV-aAg concentration in human serum by virtue of polystyrene microspheres |
| CN102749456A (en) * | 2012-06-26 | 2012-10-24 | 博奥赛斯（天津）生物科技有限公司 | Kit for chemilumineseent quantitative immunoassay of angiotensin I and preparation method thereof |
| CN102798718A (en) * | 2012-08-31 | 2012-11-28 | 福建省洪诚生物药业有限公司 | Previous S1 antigen detection kit for hepatitis B virus and preparation method of previous S1 antigen detection kit |
Non-Patent Citations (2)
| Title |
|---|
| 李晓霞: "增强化学发光免疫分析的研究进展", 《延安大学学报(自然科学版)》, vol. 21, no. 4, 30 December 2002 (2002-12-30) * |
| 梁雁 等: "化学发光法测定辣根过氧化物酶浓度的实验条件探讨", 《同济医科大学学报》, vol. 27, no. 6, 31 December 1998 (1998-12-31) * |
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