CN102735849A - Enzymatic chemiluminescence immunoassay method for human heart-type fatty acid binding protein and reagent kit - Google Patents
Enzymatic chemiluminescence immunoassay method for human heart-type fatty acid binding protein and reagent kit Download PDFInfo
- Publication number
- CN102735849A CN102735849A CN2012102337843A CN201210233784A CN102735849A CN 102735849 A CN102735849 A CN 102735849A CN 2012102337843 A CN2012102337843 A CN 2012102337843A CN 201210233784 A CN201210233784 A CN 201210233784A CN 102735849 A CN102735849 A CN 102735849A
- Authority
- CN
- China
- Prior art keywords
- fatty acid
- binding protein
- acid binding
- people
- cardiac muscle
- 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.)
- Pending
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of immunoassay analysis and particularly provides an enzymatic chemiluminescence immunoassay method for a human heart-type fatty acid binding protein. The method comprises the steps of forming solid phase-antibody-antigen-enzyme labelled secondary antibody immune sandwich complexes and detecting the immune sandwich complexes with the method of chemiluminescence. The invention also provides an enzymatic chemiluminescence immunoassay reagent kit for the human heart-type fatty acid binding protein. The detection method and the detection reagent kit are suitable for detecting and analyzing the human heart-type fatty acid binding protein and have the advantages of large detection range, high sensitivity and high specificity.
Description
Technical field
The invention belongs to the immune detection analysis technical field, a kind of people's cardiac muscle fatty acid binding protein enzymatic chemical luminous immune detection method and kit specifically are provided, be applicable to the enzyme-catalyzed chemical luminescence check and analysis of people's cardiac muscle fatty acid binding protein.
Background technology
People's cardiac muscle fatty acid binding protein (FABP) is a fatty acid binding protein in a kind of important cell.Its molecular weight is less to be the plasmosin of 15kD, is present in a large number in the cardiac muscular tissue, and its content in cardiac muscle is higher 10 times than skeletal muscle, very low at kidney, liver, small intestine content.It is intervened in the cardiac muscle cell in the cell of free fatty acid and transports, and the cardiac muscle cell is provided energy.Myocardial ischemia causes myocardium cell necrosis, and h-FABP leaks into the blood from the cardiac muscle cell, thereby the h-FABP of peripheral blood is risen.The biochemical marker that raising in the blood in the 6h of back appears in AMI is the early sign thing; Back 1～2h takes place at AMI and raises immediately in h-FABP, and continues to 12h, keeps higher level and higher susceptibility; About 24h, recover and, become the early stage biochemical marker of desirable diagnosis of AMI near normal level.
At present the detection method of people's cardiac muscle fatty acid binding protein mainly contain enzyme-linked immunosorbent assay (Enzyme-linked immunosorbent assay, ELISA) and colloidal gold immunity chromatography (Gold-immunochromatography assay, GICA).Advantages such as that the ELISA method has is simple to operate, the technology reliable, reagent conveniently is easy to get, but its susceptibility, specificity are not high enough, not accurate enough as quantivative approach; The GICA method is easy, quick, can single part or measure in batch, but its sensitivity more is lower than the ELISA method, and generally can only be as quilitative method, and can not be as quantivative approach.
Chemiluminescence (ChemiLuminescence abbreviates CL as) analytic approach is a type in the mulecular luminescence spectrographic method, and its principle is the unsettled excited state intermedium that in chemical reaction, generates, and when it gets back to ground state, discharges photon.Chemiluminometry mainly is the principle that is linear quantitative relation according to the chemiluminescence intensity of testing concentration and system in the chemical detection system under certain condition; Utilize the detection of instrument, and confirm a kind of trace analysis method of determinand content the system chemiluminescence intensity.In immune detection is analyzed, use chemiluminescence, can make the scope of detection reach 6 one magnitude, and sensitivity is very high, reaches 10
-18Mol level adds that label is stable, and the term of validity is long, makes it receive increasing concern and application.
But still do not utilize chemiluminometry to measure the research and the application of people's cardiac muscle fatty acid binding protein at present.
Summary of the invention
In order further to improve susceptibility, specificity and the accuracy that people's cardiac muscle fatty acid binding protein detects; The inventor is applied to the enzyme-catalyzed chemical luminescence analytical approach in the detection of people's cardiac muscle fatty acid binding protein; Through a large amount of experiments; Find out a kind of sensitivity, accurate and stable detection method, accomplished the present invention thus.
One aspect of the invention relates to a kind of people's cardiac muscle fatty acid binding protein detection method, and it may further comprise the steps:
(1) the anti-people's cardiac muscle of immobilization fatty acid binding protein antibody is fixed on solid phase surface, adds anti-people's cardiac muscle fatty acid binding protein antibody of sample to be tested and mark, incubation washs with cleansing solution then; Randomly, the contrast of setter's H-FABP standard items simultaneously;
(2) add the chemical luminous substrate working fluid, place a period of time;
(3) measure luminous value, obtain people's cardiac muscle fatty acid binding protein concentration value of sample to be tested.
Wherein the incubation in the step (1) does, under 18-42 ℃ of condition, is preferably under the 20-37 ℃ of condition, and incubation 5-120min is preferably incubation 10-60min, 15-45min more preferably, in one embodiment of the invention, and under 37 ℃ of conditions, incubation 30min.
Wherein the placement in the step (2) is put a period of time and is meant 1-30min, is preferably 2-20min.In one embodiment of the invention, be 5min standing time.
Particularly, detection method is following:
1. in the polystyrene micropore plate that is fixed with anti-people's cardiac muscle fatty acid binding protein monoclonal antibody, add 5-100 μ L serum or plasma sample, standard items, mixing; The anti-people's cardiac muscle fatty acid binding protein monoclonal antibody that adds 5-100 μ L horseradish peroxidase-labeled, 37 ℃ incubation 10-60 minute.In one embodiment of the invention, incubation 30min.
2. washing: can adopt hand washing or wash plate machine washing and wash.Manual manipulation method is: directly get rid of and encapsulate the liquid in the plate, and add cleansing solution (300 μ L/ hole), pour out cleansing solution and bounce to remove wall built-up liquid, triplicate is washed the plate machine operation and is seen the instrument instructions.
3. add the chemical luminous substrate working fluid: every hole adds 50-400 μ L, and said luminous substrate working fluid comprises substrate solution, auxiliary light emission solution and increased response agent.Measured in 2-20 minute behind the mixing.
4. read luminous value: the luminous value of on the luminescence assays appearance, measuring every hole.
Another aspect of the present invention relates to people's cardiac muscle fatty acid binding protein detection kit, and it comprises: be connected with the microwell plate of anti-people's cardiac muscle fatty acid binding protein antibody, the anti-people's cardiac muscle fatty acid binding protein antibody and the chemical luminous substrate working fluid of mark.
In the present invention, said chemical luminous substrate working fluid comprises substrate solution, auxiliary light emission solution and increased response agent.
In the present invention, said kit can also comprise cleansing solution, people's cardiac muscle fatty acid binding protein standard items and quality controling serum.
In the present invention, sample to be tested can be serum, blood plasma, and in one embodiment of the invention, sample to be tested is a serum.
In the present invention; Described immobilised anti-people cardiac muscle fatty acid binding protein antibody be used for anti-people's cardiac muscle fatty acid binding protein antibody of mark and be monoclonal antibody or polyclonal antibody independently of one another; In one embodiment of the invention, anti-people's cardiac muscle fatty acid binding protein antibody of immobilised anti-people's cardiac muscle fatty acid binding protein antibody and mark (enzyme mark) is monoclonal antibody;
Wherein immobilised anti-people's cardiac muscle fatty acid binding protein antibody can be identical with the myocardium fatty acid binding protein antibody of the anti-people of mark, also can be inequality.In one embodiment of the invention, anti-people's cardiac muscle fatty acid binding protein antibody of described immobilised anti-people's cardiac muscle fatty acid binding protein antibody and mark is available from Finland hytest company.In a specific embodiments of the present invention; The antibody of described immobilised antibody and mark is anti-people's cardiac muscle fatty acid binding protein monoclonal antibody of company of Finland hytest company; The clone number is 28 and 22, and its corresponding article No. is respectively 4F29 (28) and 4F29 (22).In another specific embodiments of the present invention; The antibody of described immobilised antibody and mark is anti-people's cardiac muscle fatty acid binding protein monoclonal antibody of Finland hytest company; The clone number is 28 and 31, and its corresponding article No. is respectively 4F29 (28) and 4F29 (31), for each above-mentioned antagonist; When one of them during as immobilization antibody, another is the antibody of mark.
Said people's cardiac muscle fatty acid binding protein antibody (antibody that comprises immobilization and mark) can be the antibody that utilizes animal preparation commonly used in the immunology; For example mouse-anti people cardiac muscle fatty acid binding protein antibody, the anti-people's cardiac muscle fatty acid binding protein antibody of rabbit or goat-anti people cardiac muscle fatty acid binding protein antibody; In one embodiment of the invention, be mouse-anti people cardiac muscle fatty acid binding protein antibody.
In the present invention, the label of the anti-people's cardiac muscle of described mark fatty acid binding protein antibody is a peroxidase, for example is horseradish peroxidase or Tobacco Peroxidase; Said substrate can be by the material of said enzyme stimulated luminescence under certain condition; In one embodiment of the invention, said label is a horseradish peroxidase, and said substrate is the luminol or derivatives thereof.
Said luminol derivant includes but not limited to the different Shandong promise of basic N one ethyl of different luminol, 4-amino, and N-(6-amino base)-different luminol of N-ethyl (AHEI) and N-(4-amino butyl)-different luminol of N-ethyl (ABEI).
In the present invention, the solution of said auxiliary light emission includes but not limited to superoxol, potassium ferricyanide solution, iodide solution, liquor potassic permanganate, in one embodiment of the invention, is superoxol; Said increased response agent be for can strengthen the luminous material of substrate, and when substrate was the luminol or derivatives thereof, the increased response agent included but not limited to iodophenol, p bromophenol, in one embodiment of the invention, and for to iodophenol.
Substrate solution is luminous under the effect of auxiliary light emission solution and increased response agent, and luminous intensity is directly proportional with the content of people's cardiac muscle fatty acid binding protein in the sample.
In the present invention; Cleansing solution is for removing the liquid of non-specific binding between antigen-antibody; For example phosphate buffer (PBS solution) or Tri(Hydroxymethyl) Amino Methane Hydrochloride (Tris-HCl) damping fluid also contain Tween-20 simultaneously, and the concentration of Tween-20 is 0.1-1 ‰; Be preferably 0.2-0.8 ‰, more preferably 0.3-0.6 ‰.In one embodiment of the invention, the composition of cleansing solution (1000ml) comprises sodium chloride (NaCl) 2g, sodium dihydrogen phosphate (NaH
2P0
4.12H
20) 2.9g, dipotassium hydrogen phosphate (K
2HP0
4) 0.2g, potassium chloride (KCl) 0.2g, triton x-100 0.5mL, Tween-20 0.5mL, Blanc Ni Daokesi (Bronidox-L) 5g.
In the present invention, will resist people's cardiac muscle fatty acid binding protein monoclonal antibody to be connected polystyrene micropore plate as solid-phase reagent.Described antibody is connected with microwell plate, can be directly, also can be indirect (as passing through biotin-avidin system etc.); Can connect through physisorption (electrostatic interaction, ionic link or hydrophobic effect etc.) or chemical coupling.
Described labelled antibody coupling method is to use sodium periodate activation horseradish peroxidase, with the mixed of antibody with 1:2 to 2:1, and uses the gel chromatography enzyme labelled antibody.
In the present invention, the anti-people's cardiac muscle fatty acid binding protein antibody that after adding sample to be tested, adds is ELIAS secondary antibody, and the anti-people's cardiac muscle fatty acid binding protein antibody that is connected with the microwell plate solid phase is one to resist.
After adding ELIAS secondary antibody, promptly form the immune sandwich complex of solid phase-antibody-antigen-ELIAS secondary antibody.
The beneficial effect of the invention
1, use two monoclonal antibodies, high specificity has been avoided the influence of chaff interference, makes immunoreactive affinity higher, and the production differences between batches of monoclonal antibody are relatively little, guarantee more easily product batch between stable.
2, use chemical luminous substrate solution, the sensitivity of detection and specificity are improved, and the range of linearity is wideer.
3, the antibody after the coupling of use solid phase, the antigen or the antibody that can use physical method for separation to combine have reduced non-specific binding.
4, people's cardiac muscle fatty acid binding protein detection method provided by the invention and kit; Sensitivity, specificity that people's cardiac muscle fatty acid binding protein is detected are higher; The pathology of just can the auxiliary judgment examiner whether suffering from myocardial necrosis in conjunction with other clinical symptoms; Like acute myocardial ischemia, miocardial infarction etc.
Description of drawings
Fig. 1 is for detecting the canonical plotting of FABP concentration and luminous value.
Wherein horizontal ordinate is a FABP concentration, and unit is μ g/L; Ordinate is a luminous value, and unit is RLU (relative luminous intensity).
Embodiment
The invention discloses a kind of people's cardiac muscle fatty acid binding protein enzymatic chemical luminous immune detection method and kit, those skilled in the art can use for reference this paper content, suitably improve technological parameter and realize.Special needs to be pointed out is that all similarly replace and change apparent to those skilled in the art, they all are regarded as and are included in the present invention.Method of the present invention, kit are described through preferred embodiment; The related personnel obviously can change or suitably change and combination methods and applications as herein described in not breaking away from content of the present invention, spirit and scope, realizes and use technology of the present invention.
To combine embodiment that embodiment of the present invention are described in detail below, but it will be understood to those of skill in the art that the following example only is used to explain the present invention, and should not be regarded as limiting scope of the present invention.Unreceipted actual conditions person among the embodiment carries out according to the condition of normal condition or manufacturer's suggestion.The unreceipted person of production firm of agents useful for same or instrument, being can be through the conventional products of commercial acquisition.
In the present invention; Anti-people's cardiac muscle fatty acid binding protein antibody of described immobilised anti-people's cardiac muscle fatty acid binding protein antibody and mark is available from company of Finland hytest company; It is anti-people's cardiac muscle fatty acid binding protein monoclonal antibody; Wherein a pair of clone number is 28 and 22, and its corresponding article No. is respectively 4F29 (28) and 4F29 (22); Another number is 28 and 31 to the clone, and its corresponding article No. is respectively 4F29 (28) and 4F29 (31).For each above-mentioned antagonist, when one of them during as immobilization antibody, another is the antibody of mark.
The preparation of embodiment 1 horseradish peroxidase (HRP) labelled antibody
To resist people's cardiac muscle fatty acid binding protein monoclonal antibody, concentration is concentrated into 2mg/mL.Use the sodium periodate method that improved to carry out mark, concrete steps are:
(1) 5mg HRP (available from U.S. sigma company) is dissolved in the 0.5ml distilled water, adds the 0.06Mol/L NaIO of new preparation
4WS 0.5ml, mixing is put 4 ℃, 30min;
(2) add 0.16Mol/L glycol water 0.5ml, room temperature is placed 30min;
(3) add the WS 2.5ml contain the anti-people's cardiac muscle of 5mg purifying fatty acid binding protein monoclonal antibody (clone number 22), mixing, and the bag filter of packing into slowly stir dialysis 6h (or spending the night) to 0.05Mol/L pH 9.5 carbonate buffer solutions, make it to combine;
(4) take out solution in the bag filter, add NaBH
4Solution (5mg/ml) 0.2ml, mixing is put 4 ℃, 2h;
(5) in above solution, slowly add isopyknic saturated ammonium sulfate solution, mixing, 4 ℃ of 30min; Centrifugal, remove supernatant, deposition is with a little 0.02Mol/L pH7.4PBS solution dissolving; The bag filter of packing into, spends the night at 4 ℃ of dialysis desalinations with 0.02Mol/L pH7.4PBS liquid;
(6) take out solution in the bag filter next day, centrifugal, to remove insolubles, promptly get enzyme-antibody (HRP-IgG) bond (antibody of HRP mark), add to 5ml with 0.02Mol/L pH7.4PBS liquid.Use Superdex200 gel chromatography column separating purification then, collect first and second peaks, remove the free antibodies and the enzyme that do not connect, connector is stored in 4 ℃.Use contains the Tris-HCI damping fluid (pH7.4) of 0.1% bovine serum albumin(BSA), two of HRP mark is resisted be diluted to 1.0ug/mL, as working fluid.
The adsorption of immobilization of embodiment 2 coated antibodies
Use 0.02M; The anti-people's cardiac muscle of PH7.4 phosphate buffer (PBS solution) dilution fatty acid binding protein monoclonal antibody (clone number 28) is to 2.5 μ g/ml; The microwell plate that adds polystyrene, 100 μ L/ holes, 4 ℃ are spent the night; Clean three times with cleansing solution, clap dried subsequent use after 2 hours with the 300 μ L/ hole normal temperature sealings of 1% bovine serum albumin(BSA).
The cleansing solution compound method is: sodium chloride (NaCl) 2g, sodium dihydrogen phosphate (NaH
2P0
4.12H
20) 2.9g, dipotassium hydrogen phosphate (K
2HP0
4) 0.2g, potassium chloride (KCl) 0.2g, triton x-100 0.5mL, Tween-20 0.5mL, Blanc Ni Daokesi (Bronidox-L) 5g is made into 1000mL with purified water, filters with 0.2 μ m filtrator, in room temperature preservation.
The affine immobilization of embodiment 3 coated antibodies
Use 0.02M, PH7.4 phosphate buffer dilution Avidin to 1.0 μ g/ml adds in the polystyrene micropore plate; 100 μ L/ holes, 4 ℃ are spent the night, and clean three times with cleansing solution (compound method is with embodiment 2); (the biotin labeling method is referring to the operation instruction of
Sulfo-NHS-Biotin of Thermofisher to add biotin labeled anti-people cardiac muscle fatty acid binding protein antibody (clone number 22); Article No. is 21217), concentration 1.0 μ g/ml; 100 μ L/ holes; 4 ℃ are spent the night, and clean three times with cleansing solution, clap dried subsequent use after 2 hours with the 300 μ L/ hole normal temperature sealings of 1% bovine serum albumin(BSA).
The enzyme-catalyzed chemical luminescence immune detection of embodiment 4 people cardiac muscle fatty acid binding protein
Material and equipment:
1, people's cardiac muscle fatty acid binding protein standard items 0.5-50 μ g/L.
Its compound method is: 2g sodium chloride, 2.42g trihydroxy aminomethane, 5g bovine serum albumin(BSA), the general labor of 0.5mL Kelin 300.With the dissolving of 1000mL purified water, transfer pH to 7.6 with hydrochloric acid, 0.2 μ m filtrator filters, in 4 ℃ of preservations, as the damping fluid of standard items dilution.Confirm the pure article concentration of people's cardiac muscle fatty acid binding protein according to pierce protein quantification detectable BCA method, use damping fluid to dilute (the standard items scope is 0.5-50 μ g/L) then,, be used for the production standard curve in 4 ℃ of preservations.
2, antibody solid-phase coating microwell plate: specifically see embodiment 2.
3, horseradish peroxidase (HRP) labelled antibody: specifically see embodiment 1.
Enzyme labelled antibody is used diluted, and the diluent preparation method adds 0.5% general labor Kelin 300 for using calf serum, filters with 0.45 μ m filtrator.
4, cleansing solution 100mL, embodiment 2 is seen in preparation.
5, chemical luminous substrate working fluid, referring to patent " supersensitive enzyme accelerator for chemical luminescence for liquid ", the patent No. is ZL91110621.9; Its concrete prescription is the 1.25mmol/L luminol, 0.136mmol/L is right-and iodophenol, 10mmol/L TrisHCL (pH8.6); 0.2% ethanol; 0.3mmol/LNaCl, 5mmol/L cyclohexanediaminetetraacetic acid (CDTA), the H of 4mmol/L
2O
2NaBO with 4mmol/L
3Mix.
6, the patients serum 100 parts (wherein 50 routine clinical diagnosises are miocardial infarction, and 50 examples are myocardial ischemia, 100 parts of normal human serums.
7, MPC-1 chemical luminescence detector (source, Beijing moral bioengineering company limited).
8, water bath (being used for 37 ℃ of temperature bathes).
Operation steps:
1, application of sample and immune response: in the antibody solid-phase coating microwell plate of embodiment 2 preparations, add 20 μ L patients, normal serum sample or people's cardiac muscle fatty acid binding protein standard items; The enzyme labelled antibody working fluid 100 μ L that in each micropore, add embodiment 1 preparation; Behind the mixing, 37 ℃ of incubations 30 minutes.
2, washing: every hole adds 300 μ L cleansing solutions, dries then.Repeat twice.
3, add the chemical luminous substrate working fluid: every hole adds 100 μ L chemical luminous substrate working fluids.
4, add after substrate and the reinforcing agent mixed liquor and to read luminous value in 5 minutes: measure the luminous value in every hole at the luminescence assays appearance, minute was 1 second.
5, testing result: the typical curve that detects FABP concentration and luminous value is seen Fig. 1.
Zero standard is carried out 20 repeated tests on schedule, get the mean value that zero standard measures on schedule and add 2 times standard deviation, be its sensitivity.The sensitivity of this method is＜0.5 μ g/L.
200 routine clinical serum are measured; Be limited to 1.56 μ g/L on the normal range of chemiluminescence determination; Can obtain the result in the table 1 according to this criterion; Simultaneously the detection method of colloidal gold immunity chromatography and enzyme-linked immunosorbent assay method carries out according to separately instructions that (colloidal gold kit is available from Shenzhen Kang Shengbao Bioisystech Co., Ltd, and article No. is A207002; The ELISA detection kit is available from Dutch hycult company, and article No. is HK402).
Testing result is seen table 1.
Each method of table 1. is for the situation that detects of 200 parts of clinical serum
Can find out that from above result compare with enzyme-linked immunosorbent assay with colloidal gold immunity chromatography, chemical luminous immune detection method of the present invention has higher sensitivity and specificity.
In the present embodiment, anti-people's cardiac muscle fatty acid binding protein antibody of employed immobilised anti-people cardiac muscle fatty acid binding protein antibody and enzyme labeling is respectively that to clone number be 28 and 22 antibody; The inventor proves, and the place-exchange of two kinds of antibody (clone who is the antibody of immobilization and enzyme labeling number is respectively 22 and 28) also can be obtained identical detection effect.Simultaneously, the inventor proves also and uses clone numbers 28 also can reach identical detection effect with an antagonist of 31 that its position also can exchange.
Although embodiment of the present invention has obtained detailed description, it will be understood to those of skill in the art that.According to disclosed all instructions, can carry out various modifications and replacement to those details, these change all within protection scope of the present invention.Four corner of the present invention is provided by accompanying claims and any equivalent thereof.
Claims (12)
1. people cardiac muscle fatty acid binding protein detection method, it may further comprise the steps:
(1) the anti-people's cardiac muscle of immobilization fatty acid binding protein antibody is fixed on solid phase surface, adds anti-people's cardiac muscle fatty acid binding protein antibody of sample to be tested and mark, incubation washs with cleansing solution then; Randomly, the contrast of setter's H-FABP standard items simultaneously;
(2) add the chemical luminous substrate working fluid, place a period of time;
(3) measure luminous value, obtain people's cardiac muscle fatty acid binding protein concentration value of sample to be tested.
2. the detection method of claim 1, the myocardium fatty acid binding protein antibody of anti-people of wherein said immobilization anti-people's cardiac muscle fatty acid binding protein antibody and mark is monoclonal antibody or polyclonal antibody independently of one another, is preferably monoclonal antibody; Anti-people's cardiac muscle fatty acid binding protein antibody of anti-people's cardiac muscle fatty acid binding protein antibody of described immobilization and mark can be identical, also can be inequality.
3. the detection method of claim 1, wherein said label is a superoxide, is preferably horseradish peroxidase or Tobacco Peroxidase.
4. the detection method of claim 3, wherein said substrate is the luminol or derivatives thereof.
5. the detection method of claim 1, wherein the incubation in the step (1) does, 18-42 ℃, be preferably 20-37 ℃, for example be under 37 ℃ of conditions, incubation 5-120min is preferably incubation 10-60min, more preferably 15-45min.
6. the detection method of claim 1, wherein the cleansing solution in the step (1) is phosphate buffer or Tri(Hydroxymethyl) Amino Methane Hydrochloride damping fluid, also contains Tween-20 simultaneously, and its concentration is 0.1-1 ‰, is preferably 0.2-0.8 ‰, and more preferably 0.3-0.6 ‰.
7. the detection method of claim 1, wherein the placement a period of time in the step (2) is 1-30min, is preferably 2-20min.
8. people cardiac muscle fatty acid binding protein detection kit; It comprises: the microwell plate that is connected with the anti-people's cardiac muscle of immobilization fatty acid binding protein antibody; The anti-people's cardiac muscle fatty acid binding protein antibody and the chemical luminous substrate working fluid of mark, said chemical luminous substrate working fluid comprises substrate solution, auxiliary light emission solution and increased response agent.
9. the kit of claim 8, it also comprises cleansing solution and people's cardiac muscle fatty acid binding protein standard items.
10. the kit of claim 8, the myocardium fatty acid binding protein antibody of anti-people of wherein said immobilization anti-people's cardiac muscle fatty acid binding protein antibody and mark is monoclonal antibody or polyclonal antibody independently of one another, is preferably monoclonal antibody; Anti-people's cardiac muscle fatty acid binding protein antibody of anti-people's cardiac muscle fatty acid binding protein antibody of described immobilization and mark can be identical, also can be inequality.
11. the kit of claim 8, wherein said label is a superoxide, is preferably horseradish peroxidase or Tobacco Peroxidase.
12. the kit of claim 11, wherein said substrate are the luminol or derivatives thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102337843A CN102735849A (en) | 2012-07-05 | 2012-07-05 | Enzymatic chemiluminescence immunoassay method for human heart-type fatty acid binding protein and reagent kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102337843A CN102735849A (en) | 2012-07-05 | 2012-07-05 | Enzymatic chemiluminescence immunoassay method for human heart-type fatty acid binding protein and reagent kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102735849A true CN102735849A (en) | 2012-10-17 |
Family
ID=46991717
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012102337843A Pending CN102735849A (en) | 2012-07-05 | 2012-07-05 | Enzymatic chemiluminescence immunoassay method for human heart-type fatty acid binding protein and reagent kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102735849A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103901216A (en) * | 2014-04-22 | 2014-07-02 | 上海科华生物工程股份有限公司 | Human H-FABP colloidal gold test paper and preparation method thereof |
| CN108445222A (en) * | 2018-02-01 | 2018-08-24 | 浙江艾明德生物科技有限公司 | A kind of kit and preparation method quantitatively detecting cardic fatty acid binding protein |
| CN111289758A (en) * | 2020-03-10 | 2020-06-16 | 苏州翊讯生物科技有限公司 | Kit for H-FABP quantitative detection and H-FABP quantitative detection method |
| CN111487413A (en) * | 2019-01-29 | 2020-08-04 | 艾维可生物科技有限公司 | Detection kit for quantitatively detecting heart-type fatty acid binding protein by E L ISA method |
| CN111505303A (en) * | 2019-01-31 | 2020-08-07 | 艾维可生物科技有限公司 | Kit for detecting heart-type fatty acid binding protein by chemiluminescence method and use method thereof |
| CN112326636A (en) * | 2020-10-26 | 2021-02-05 | 通山县金瑞生物科技研发中心 | High-sensitivity ECL kit and preparation method thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998057171A1 (en) * | 1997-06-13 | 1998-12-17 | Medical Biology Institute | Method of detection of cardiac ischemia using fatty acid binding protein |
| CN1396453A (en) * | 2002-05-17 | 2003-02-12 | 胡美浩 | Reagent kit for early diagnosis of acute myocardial infarction |
| CN1743411A (en) * | 2004-08-30 | 2006-03-08 | 北京源德生物医学工程有限公司 | Sensitivity-reinforced chemical light-emitting liquid containing auxiliary intensifier and intensifier |
| CN101377501A (en) * | 2007-08-31 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Cell keratin 19 fragments chemiluminescence immune analysis determination reagent kit and preparing method thereof |
| CN102226811A (en) * | 2011-04-02 | 2011-10-26 | 吉林雅强医疗器械有限公司 | Preparation method of diagnostic test paper for detecting H-FABP (heart-type fatty acid binding protein) |
| CN102399752A (en) * | 2011-10-28 | 2012-04-04 | 南京医科大学第一附属医院 | Heart type-fatty acid binding protein monoclonal antibody mcf1 and application thereof |
| CN102520194A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same |
-
2012
- 2012-07-05 CN CN2012102337843A patent/CN102735849A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998057171A1 (en) * | 1997-06-13 | 1998-12-17 | Medical Biology Institute | Method of detection of cardiac ischemia using fatty acid binding protein |
| CN1396453A (en) * | 2002-05-17 | 2003-02-12 | 胡美浩 | Reagent kit for early diagnosis of acute myocardial infarction |
| CN1743411A (en) * | 2004-08-30 | 2006-03-08 | 北京源德生物医学工程有限公司 | Sensitivity-reinforced chemical light-emitting liquid containing auxiliary intensifier and intensifier |
| CN101377501A (en) * | 2007-08-31 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Cell keratin 19 fragments chemiluminescence immune analysis determination reagent kit and preparing method thereof |
| CN102226811A (en) * | 2011-04-02 | 2011-10-26 | 吉林雅强医疗器械有限公司 | Preparation method of diagnostic test paper for detecting H-FABP (heart-type fatty acid binding protein) |
| CN102399752A (en) * | 2011-10-28 | 2012-04-04 | 南京医科大学第一附属医院 | Heart type-fatty acid binding protein monoclonal antibody mcf1 and application thereof |
| CN102520194A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same |
Non-Patent Citations (1)
| Title |
|---|
| 蒋佩军: "心肌型脂肪酸结合蛋白在非ST段抬高的心肌梗死早期诊断的临床意义", 《实用医学杂志》, vol. 25, no. 8, 31 December 2009 (2009-12-31), pages 1256 - 1257 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103901216A (en) * | 2014-04-22 | 2014-07-02 | 上海科华生物工程股份有限公司 | Human H-FABP colloidal gold test paper and preparation method thereof |
| CN103901216B (en) * | 2014-04-22 | 2015-10-21 | 上海科华生物工程股份有限公司 | People H-FABP colloidal gold test and preparation method thereof |
| CN108445222A (en) * | 2018-02-01 | 2018-08-24 | 浙江艾明德生物科技有限公司 | A kind of kit and preparation method quantitatively detecting cardic fatty acid binding protein |
| CN111487413A (en) * | 2019-01-29 | 2020-08-04 | 艾维可生物科技有限公司 | Detection kit for quantitatively detecting heart-type fatty acid binding protein by E L ISA method |
| CN111505303A (en) * | 2019-01-31 | 2020-08-07 | 艾维可生物科技有限公司 | Kit for detecting heart-type fatty acid binding protein by chemiluminescence method and use method thereof |
| CN111289758A (en) * | 2020-03-10 | 2020-06-16 | 苏州翊讯生物科技有限公司 | Kit for H-FABP quantitative detection and H-FABP quantitative detection method |
| CN111289758B (en) * | 2020-03-10 | 2024-02-09 | 苏州翊讯生物科技有限公司 | Kit for quantitative detection of H-FABP and method for quantitative detection of H-FABP |
| CN112326636A (en) * | 2020-10-26 | 2021-02-05 | 通山县金瑞生物科技研发中心 | High-sensitivity ECL kit and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1618383B1 (en) | Membrane strip biosensor system for point-of-care testing | |
| Yolken | Enzyme immunoassays for the detection of infectious antigens in body fluids: current limitations and future prospects | |
| CN104634980B (en) | The super quick detection kit of cardiac muscle troponin I and super quick detection method | |
| CN103033627A (en) | Myoglobin enzymatic chemiluminescence immunodetection method and kit | |
| CN102735849A (en) | Enzymatic chemiluminescence immunoassay method for human heart-type fatty acid binding protein and reagent kit | |
| CN204165984U (en) | Cardiac muscle troponin I immunochromatographiassay assay quantitative detection test paper | |
| CN102998467B (en) | β human chorionic gonadotrophin magnetic microparticle chemiluminescence immune quantitative detection reagent box and preparation method thereof | |
| CN104849469A (en) | Kit for detecting NGAL content and preparation method thereof | |
| CN108333357A (en) | A kind of magnetic bead time-resolved fluoroimmunoassay quantitatively detects CK-MB kits | |
| JP2006058295A (en) | Assay | |
| CN107831163A (en) | A kind of chemiluminescence detection kit of thyroglobulin and preparation method thereof | |
| CN102998465B (en) | Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for angiotensin (Ang) I, and preparation method of kit | |
| JP2005510234A (en) | Polypeptide quantification | |
| CN102520195A (en) | Chromogranin A chemiluminescence immunoassay reagent kit and preparation method thereof | |
| CN102998442A (en) | Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for aldosterone, and preparation method of kit | |
| CN102809651A (en) | Creatine phosphokinase isozyme chemiluminescence immune quantitative detection kit and preparation method thereof | |
| CN101571546A (en) | Human adiponectin enzyme-linked immunosorbent detection kit, preparation method and application thereof | |
| CN106198997B (en) | A kind of kit of Quantitative detection troponin and creatine kinase isozyme | |
| CN102998466A (en) | Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for growth hormone (GH), and preparation method of kit | |
| CN109187971A (en) | Neuronspecific enolase chemiluminescence immune detection reagent kit and preparation method thereof | |
| CN101363861A (en) | Hepatitis b virus surface antigen chemiluminescence immune assay determination kit and method for preparing same | |
| CN105628930A (en) | Troponin I detection reagent with high sensitivity through latex enhanced turbidimetric Immunoassay | |
| CN101377509A (en) | III type precollagen N end peptide chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof | |
| CN102998462B (en) | Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for procollagen III (PC III), and preparation method of kit | |
| CN108152486A (en) | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of human soluble ST2 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121017 |