CN105759055A - P1NP detection kit and preparation method thereof - Google Patents
P1NP detection kit and preparation method thereof Download PDFInfo
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- CN105759055A CN105759055A CN201610207318.6A CN201610207318A CN105759055A CN 105759055 A CN105759055 A CN 105759055A CN 201610207318 A CN201610207318 A CN 201610207318A CN 105759055 A CN105759055 A CN 105759055A
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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
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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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
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Abstract
The invention discloses a detection kit for I-type collagen amino terminal extending peptide (P1NP) in human serum or plasma. The detection kit is designed on the basis of double-antibody method principle. The streptavidin coated with each pore of a porous plate pre-coated with streptavidin is 0.24-0.36 mu g; the concentration of biotin labeling mice anti-P1NP monoclonal antibody is 0.8-1.2mu g/ml and the usage is 100ul per pore; the working concentration of HRP labeled mice anti-P1NP monoclonal antibody solution is 1:(2500-3000). Through the detection kit provided by the invention, the quick detection for I-type collagen amino terminal extending peptide in human serum or plasma can be realized, the detection cost is low and the detection sensitivity is high.
Description
Technical field
The invention belongs to medical instruments field, particularly relate to type i collagen ammonia in a kind of human serum or blood plasma
Cardinal extremity extends peptide (P1NP) detection kit and preparation method thereof.
Background technology
Type i collagen propetide (P1NP) is modal ossein, is the important component part of bone matrix, almost
Account for the 90% of total protein part.Collagen in bone is synthesized with precollagenous form by Gegenbaur's cell, front
Cracking generation precollagen amino terminal peptide (P1NP) from procollagen molecule when collagen forms collagenous fibres, therefore,
Measure its level in blood, bon e formation situation can be reflected.
What P1NP reflected is the deposition conditions of type i collagen, is therefore as a bon e formation mark.At I
In the forming process of Collagen Type VI, P1NP is released into ECS and eventually enters into blood.So, in serum
Type i collagen propetide level is reflection Gegenbaur's cell activity and bon e formation within the specific limits and reflects type i collagen
The specific parameters of synthesis rate.
On the other hand, there is many defects in existing detection means: testing cost is higher, in-convenience in use,
Cannot realize quickly detecting, the sensitivity of test also ratio is relatively low.
Summary of the invention
An object of the present invention is to provide type i collagen aminoterminal in a kind of human serum or blood plasma and extends peptide reagent
Box.
Realize above-mentioned purpose technical scheme as follows:
In a kind of human serum or blood plasma, type i collagen aminoterminal extends peptide (P1NP) detection kit, mainly wraps
Include:
1) being coated with the microwell plate of Streptavidin, each micropore coated Streptavidin amount is 0.24-
0.36μg；
2) biotin labeled mouse-anti P1NP monoclonal antibody solution, the wherein concentration of P1NP monoclonal antibody
For 0.8-1.2 μ g/ml, consumption is every micropore 100 μ l；
3) the mouse-anti P1NP monoclonal antibody solution of HRP mark, wherein the mouse-anti P1NP Dan Ke of HRP mark
The working concentration of grand antibody is 1:2500-3000.
Wherein in an embodiment, each micropore coated Streptavidin amount for 0.3 μ g.
Wherein in an embodiment, it is 1.0 μ that biotin labeled mouse-anti P1NP monoclonal antibody obtains concentration
g/ml。
Another object of the present invention is to provide the preparation method of a kind of P1NP detection kit.
The technical scheme realizing above-mentioned purpose is:
The preparation method of a kind of P1NP detection kit, mainly comprises the steps that
1) microwell plate of Streptavidin it is coated with:
In the micropore of each microwell plate, addition 80-120 μ l concentration is the Streptavidin of 3 μ g/ml, 37
± 2 DEG C are dried, and seal and preserve, wash plate, close；
2) biotin labeled mouse-anti P1NP monoclonal antibody solution is prepared: by mouse-anti P1NP antibody PBS
Buffer solution and EDTA are diluted to final concentration of 2mg/ml, are subsequently adding NH2-Reactive Biotin makes volume
For 0.5ml, mixing is placed on Incubation in dark in 37 ± 2 DEG C of insulating boxs gently, centrifugal, then uses 10mmol/L
PBS and the EDTA of 25mol/L be diluted to final concentration of 0.8-1.2 μ g/ml.
Preferably, the amount adding Streptavidin in described each micropore is 100 μ l.
Preferably, described microwell plate wash plate and being closed as: add the next day of Streptavidin, discard in hole
Liquid, adds cleaning solution 280-300ul/ hole, rinses 3 times, each 3min；Add by 200-250ul/ hole
Confining liquid, closes 1-2 hour or closes overnight at 4 DEG C, then discard hole inner sealing liquid for 37 ± 2 DEG C.
Described P1NP detection kit also includes the mouse-anti P1NP monoclonal antibody of HRP mark, this HRP
The preparation manipulation of the mouse-anti P1NP monoclonal antibody antibody of mark is:
1) take mouse-anti P1NP monoclonal antibody freeze-dried powder 0.1g, be the PBS of 10mmol/L with 10ml concentration
Buffer solution mixes；
2) take HRP dry powder deionized water and be diluted to 5mg/ml, use simple Over-voltage protection by HRP with anti-
Body cross-links；
3) take the mouse-anti P1NP monoclonal antibody of the mark of the HRP after preparation, buffer with the PBS of 10mmol/L
The EDTA dilution of liquid and 25mol/L, the work of the mouse-anti P1NP monoclonal antibody of the HRP mark after dilution
Concentration is 1:2500-3500, preserves after adding preservative and stabilizer.
Being prepared as of described mouse-anti P1NP monoclonal antibody: take 6 small white mouses, uses injected s. c to enter
Row immunity inoculation, initial immunity injection 1mg/mL P1NP antigen 0.2ml/ only, the 3rd week, the 4th week and the
5 weeks injection 1mg/mL P1NP antigen is with incomplete Freund's adjuvant equal-volume mixed solution 0.4ml/ only；5th
Take blood examination after week and survey titer, if more than 1:10000, then draw neck to take spleen after putting to death mouse, then use miscellaneous
Knurl technology is handed over to prepare monoclonal antibody.
Described P1NP detection kit uses ELISA, resists solubility P1NP based on anti-P1NP antibody
Former double-antibody method Cleaning Principle designs.Described P1NP detection kit has specifically, sensitivity and precision
Spend the most extraordinary advantage.
Detailed description of the invention
P1NP detection kit of the present invention, its detection method is: add biotin labeling in micropore
Mouse-anti P1NP monoclonal antibody solution hatch at 20-25 DEG C 30 minutes, first it be bound to pre-coated
Streptavidin in micropore.Calibration object, quality-control product and sample are incubated after joining micropore at 20-25 DEG C
Educate 60 minutes and make itself and antibody response, add the mouse-anti P1NP monoclonal antibody of HRP mark, in room temperature
Under hatch 60 minutes, then liquid washing in sucking-off hole.Add chromogenic substrate (TMB) after completing to show
Look.Being eventually adding stop buffer and read absorbance on ELIASA, color intensity is directly proportional to P1NP concentration.
Embodiment 1
P1NP detection kit described in the present embodiment to mainly comprise composition as follows:
1, ELISA Plate: be coated with Streptavidin, 12 x8 holes in micropore.
2, calibration object A-F: the phosphate buffer of the hyclone containing P1NP that can be used directly, 6 ×
0.4ml。
3, quality-control product: the phosphate buffer of the hyclone containing P1NP that can be used directly, 2 × 0.4ml.
4, biotin labeled antibody: the phosphoric acid of the protein stabiliser containing biotin labeled P1NP antibody
Salt buffer, 1 × 0.2ml.
5, enzyme conjugates: containing mouse-anti P1NP monoclonal antibody, enzyme stabilizers and the preservative of HRP mark
Phosphate buffer, 1 × 0.2ml.
6, incubation buffer: containing the phosphate buffer of protein stabiliser, 1 × 30ml.
7, washing lotion: the 50 × Tris salt buffer containing tween, 1 × 20ml.
8, substrate solution (TMB): tetramethyl benzidine (TMB) substrate that can be used directly (is dissolved in acidic buffer
In), 1 × 12ml.
9, stop buffer: 0.3M dilute sulfuric acid, 1 × 12ml.
10, shrouding film.
The preparation method of described P1NP detection kit is as follows:
1, the pre-coated microwell plate of Streptavidin:
Streptavidin outsourcing, is diluted to 3 μ g/ml with the PBS of 0.02M pH 7.4.
1) it is coated
In each micropore, add the solution of streptavidin that 0.1ml has diluted, 37 DEG C of dry 2h, add a cover for 4 DEG C
Seal and preserve.
2) plate is washed
Discard liquid in hole next day, blotting paper cramps to remove residual liquid gently, add cleaning solution (about
280-300ul/ hole), rinse 3 times, each 3min (cleaning solution: containing the PBS of 0.05%Tween 20).
3) close
Add confining liquid by 200-250ul/ hole, close 1-2 hour for 37 DEG C, it is possible to close overnight, so for 4 DEG C
After discard hole inner sealing liquid (confining liquid: 1%BSA, formulated with 10mM PBS).
2, biotin labeling mouse-anti P1NP monoclonal antibody solution is prepared
1) prepared by antibody
Take 6 small white mouses, use injected s. c to carry out immunity inoculation, initial immunity injection 1mg/mL P1NP
Antigen (outer purchased from nm crystallite bio tech ltd, Guangzhou) 0.2ml/ only, the 3rd week, the 4th week and
With incomplete Freund's adjuvant equal-volume mixed solution 0.4ml/ only within 5th week, inject 1mg/mL P1NP antigen, the
Take blood examination after 5 weeks and survey titer, if more than 1:10000, then draw neck to take spleen after putting to death mouse, then use
Hybridoma technology prepares monoclonal antibody.
2) biotin labeled mouse-anti P1NP monoclonal antibody
By PBS and the EDTA dilution of 25mol/L of mouse-anti P1NP monoclonal antibody 10mmol/L
To final concentration of 2mg/ml, it is subsequently adding 13.0ul NH2-Reactive Biotin makes volume be 0.5ml,
Mixing is placed on Incubation in dark 30min in 37 DEG C of insulating boxs gently.12,000xg is centrifuged 10min.
With dilution (10mmol/L PBS, 25mol/L EDTA), biotin labeled mouse-anti monoclonal is resisted
Body is diluted to 0.1 μ g/ml, 1.0 μ g/ml, 2.0 μ g/ml, 4.0 μ g/ml, 6.0 μ g/ml, 8.0 μ
G/ml, 10.0 μ g/ml six concentration, through integration test and assessment, obtain the OD value antibody when 1.0
Concentration is as working concentration.Finally, the concentration of the biotin labeling mouse-anti P1NP antibody determined is: 0.8-
1.2 μ g/ml, the optium concentration of the present embodiment is 1.0 μ g/ml.
3, enzymic-labelled antibody is prepared
Take mouse-anti P1NP antibody freeze-dried powder 0.1g, mix with the PBS that 10ml concentration is 10mmol/L；
Take HRP dry powder deionized water and be diluted to 5mg/ml, use simple Over-voltage protection by HRP with antibody linked；
Take the HRP-mouse-anti P1NP antibody after preparation, with PBS and the EDTA of 25mol/L of 10mmol/L
Dilution, the working concentration of HRP labelled antibody is 1:2500-3500, dispenses after adding preservative and stabilizer
Preserve.
The selection of enzymic-labelled antibody working concentration: first the standard items of concentration 3 μ g/ml are added in micropore and react,
Add after washing a series of different dilution factor (1:1000,1:2000,1:3000,1:4000,1:5000,1:
6000:, 1:7000) enzyme conjugates hatch, add after washing substrate colour developing, add stop buffer terminate reaction.
ELIASA measures its OD value, selects that enzyme conjugates dilution factor of OD value >=1.0 to be enzyme conjugates
Working concentration.The enzyme labelled antibody best effort concentration finally determining the present embodiment is 1:3000.
4, calibration object and the preparation of quality-control product
1) source: outsourcing P1NP hyclone Han 1mg/ml.
2) prepared by quality-control product
Preparation 500ml quality-control product: press 70.18ng/ml and 406.95 with the P1NP hyclone containing 1mg/ml
Prepared by ng/ml concentration.
A) 1mg/ml P1NP solution 35.1 μ l+500ml calibration object buffer solution is taken.
B) 1mg/ml P1NP solution 0.203ml+499.79ml calibration object buffer solution is taken.
3) prepared by calibration object
The calibration object of each concentration all prepares 500ml, concrete process for preparation following (joining from high concentration).
A: take containing 1mg/mlP1NP hyclone 0.78ml+999.22ml calibration object buffer solution；
B: take A calibration object 250ml+250ml calibration object buffer solution；
C: take A calibration object 125ml+375ml calibration object buffer solution；
D: take A calibration object 62.5ml+437.5ml calibration object buffer solution；
E: take A calibration object 31.25ml+468.75ml calibration object buffer solution；
F:500ml calibration object buffer solution.
The concentration of each calibration object of gained is respectively A:780ng/ml, B:390ng/ml, C:195ng/ml, D:97.5
Ng/ml, E:48.75ng/ml, F:0ng/ml.
Embodiment 2: use the detection kit described in embodiment 1 that sample is detected
Before using, by all solution equilibrias to room temperature, test at 20-25 DEG C.
Determine the required microwell plate quantity of experiment.It addition, often wheel experiment needs 16 holes for standard items and Quality Control altogether
Product.An appropriate number of microwell plate is placed on plastic frame.By untapped microwell plate together with drier close
It is encapsulated in Fresco Bag.
Concrete operation step is as follows:
1) biotin labeled mouse-anti P1NP monoclonal antibody that 100 μ l diluted is added in requirement
In ELISA Plate micropore, cover shrouding film, ELISA Plate oscillator (300 revs/min) is incubated at 20-25 DEG C
Educate 30 minutes.
2) plate is washed 5 times by the washing lotion diluted.
A) plate is automatically washed: arrange and wash trigger distribution every hole at least 250 μ l washing lotion, be repeated 5 times.
B) plate is washed by hand: pour out thing in hole rapidly.Every hole adds 250 μ l flushing liquors, gets rid of rapidly hole
Interior washing lotion, is inverted in plate on blotting paper and firmly pats to remove the washing lotion of residual.Repeat this step 4 time.
3) 50 μ l calibration objects, quality-control product and sample are added in corresponding ELISA Plate micropore, the most again with moving
Liquid device add in all micropores 50 μ l incubation buffer (note: this operation should complete in 15 minutes with
Reduce error).
4) cover shrouding film, ELISA Plate oscillator (300 revs/min) hatches 60 points at 20-25 DEG C
Clock.
5) plate is washed 5 times with the dcq buffer liquid diluted.
6) 100 μ l freshly prepared enzyme conjugates solution is added.
7) cover shrouding film, ELISA Plate oscillator (300 revs/min) hatches 60 points at 20-25 DEG C
Clock.
8) plate is washed 5 times with the dcq buffer liquid diluted.
9) in every hole, 100 μ l tmb substrates are added with Multi-channel liquid transfer device.
10) cover shrouding film, ELISA Plate oscillator (300 revs/min) hatches 15 points at 20-25 DEG C
Clock.
11) in every hole, 100 μ l stop buffers are added with Multi-channel liquid transfer device.
12), after adding stop buffer, read at ELIASA 450nm (with reference to 650nm) wavelength in 30 minutes and inhale
Luminosity.
[reference value (term of reference)]
The term of reference of various people is exemplified below.
[explanation of assay]
Use quadratic equation curve matching: y=a+b*x+c*x^2；
With absorbance as ordinate, P1NP concentration is abscissa, draws calibration curve.Count from calibration curve
Calculate the P1NP concentration value of unknown sample.
Result is illustrated:
If the absorbance of sample to be tested is higher than calibration A, it is proposed that by calibration object F diluted sample and reanalyse.
Embodiment 3: properties of product analysis
1) sensitivity
Use the P1NP detection kit described in embodiment 1, according to the detection method described in embodiment 2, enter
Row properties of product are analyzed.
Take calibration object F, by operation instructions requirement Parallel testing 10 times and record OD value, calculate average and
SD value, uses averageOD value, calculate from dosage-response curve correspondence dosage, this dosage
Value is the LDL of the method.Testing result such as following table.
According to data above, calculate mean valueBeing 0.005, standard deviation S D is 0.001, finally calculates
Low detection limit is less than 5ng/mL.
2) range of linearity
Diluted concentration is 780ng/ml, 390ng/ml, 195ng/ml, 97.5ng/ml, 48.75ng/ml, 0ng/ml
Calibration object, each calibration object detects 2 times, draws calibration curve according to kit standard method, and calculates line
Property scope.
According to data above, the range of linearity of the calibration curve measured is more than 0.99, and measurement scope is set to
5-780ng/mL is rational.
3) specific
The medicine of following variable concentrations is joined in the serum of 3 different PINP levels, carry out double detection.
Institute's mark directrix curve and maximum concentration drug test OD value are as shown in the table.
Above-mentioned experimental data shows, said medicine is the most noiseless to the rate of recovery of PINP, does not interferes with kit
Specific.
Embodiment described above is merely to illustrate a kind of detailed description of the invention when the present invention is directed to actual application,
It describes more concrete and detailed, but therefore can not be interpreted as the restriction to the scope of the claims of the present invention.Should
When it is noted that for the person of ordinary skill of the art, without departing from the inventive concept of the premise,
The some deformation made and improvement broadly fall into protection scope of the present invention.
Claims (9)
1. a people P1NP detection kit, is characterized in that, specifically includes that
(1) being coated with the microwell plate of Streptavidin, each micropore coated Streptavidin amount is 0.24
-0.36 μ g；
(2) biotin labeled mouse-anti P1NP monoclonal antibody solution, the most biotin labeled mouse-anti
The concentration of P1NP monoclonal antibody is 0.8-1.2 μ g/ml, and consumption is every micropore 100 μ l；
(3) the mouse-anti P1NP monoclonal antibody solution of HRP mark, wherein the mouse-anti P1NP of HRP mark
The working concentration of monoclonal antibody is 1:2500-3000.
P1NP detection kit the most according to claim 1, is characterized in that, described each micropore is coated
The amount of Streptavidin is 0.3 μ g.
P1NP detection kit the most according to claim 1, is characterized in that, described biotin labeled mouse
The concentration of anti-P1NP monoclonal antibody is 1.0 μ g/ml.
4. according to the P1NP detection kit described in any one of claims 1 to 3, it is characterized in that, described HRP
The working concentration of the mouse-anti P1NP monoclonal antibody of mark is 1:3000.
5. a preparation method for P1NP detection kit, is characterized in that, mainly comprises the steps that
(1) microwell plate of Streptavidin it is coated with: in the micropore of each microwell plate, add 80-120 μ l
Concentration is the Streptavidin of 3 μ g/ml, and 37 ± 2 DEG C are dried, and seal and preserve, wash plate, close；
(2) biotin labeled mouse-anti P1NP monoclonal antibody solution is prepared: by mouse-anti P1NP monoclonal
Antibody PBS and EDTA dilution, be subsequently adding NH2-Reactive Biotin, gently after mixing
It is placed in Incubation in dark in 37 ± 2 DEG C of insulating boxs, centrifugal, then it is diluted to final concentration with PBS and EDTA
For 0.8-1.2 μ g/ml.
Preparation method the most according to claim 5, is characterized in that, adds strepto-parent in described each micropore
It is 100 μ l with the amount of element.
Preparation method the most according to claim 5, is characterized in that, described microwell plate wash plate and being closed as:
Adding the next day of Streptavidin, discard liquid in hole, add cleaning solution, rinsing, by 200-250ul/
Hole adds confining liquid, closes 1-2 hour or closes overnight at 4 DEG C, then discard hole inner sealing liquid for 37 ± 2 DEG C.
Preparation method the most according to claim 5, is characterized in that, described P1NP detection kit also includes
There are the mouse-anti P1NP monoclonal antibody that HRP marks, the system of the mouse-anti P1NP monoclonal antibody of this HRP mark
Standby operation is:
(1) take mouse-anti P1NP monoclonal antibody freeze-dried powder, mix with PBS；
(2) take HRP dry powder, dilute by deionized water, use simple Over-voltage protection by HRP and mouse-anti
P1NP monoclonal antibody cross-links, and obtains the mouse-anti P1NP monoclonal antibody of HRP mark；
(3) take the mouse-anti P1NP monoclonal antibody of the mark of the HRP after preparation, dilute with PBS and EDTA,
The mouse-anti P1NP monoclonal antibody of the HRP mark after must diluting, the mouse-anti of the HRP mark after described dilution
P1NP monoclonal antibody working concentration is 1:2500-3500.
9. according to the preparation method described in any one of claim 5 to 8, it is characterized in that, described mouse-anti P1NP is mono-
Being prepared as of clonal antibody: using injected s. c to carry out immunity inoculation small white mouse, initial immunity is injected
P1NP antigen, the 3rd week, the 4th week and the 5th week injection P1NP antigen and incomplete Freund's adjuvant equal-volume
Mixed solution, takes blood examination and surveys titer after the 5th week, after immunity produces a desired effect, then after drawing neck to put to death mouse
Take spleen, then use hybridoma technology to prepare monoclonal antibody.
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| CN110317271A (en) * | 2019-07-02 | 2019-10-11 | 武汉云克隆科技股份有限公司 | Weight/the light chain variable region and encoding gene and recombinant antibodies of a kind of PINP recombinant antibodies |
| CN111007269A (en) * | 2019-11-15 | 2020-04-14 | 迪瑞医疗科技股份有限公司 | Total I type collagen amino-terminal extension peptide chemiluminescence immunoassay kit and preparation method thereof |
| CN111239384A (en) * | 2020-01-15 | 2020-06-05 | 中国人民解放军陆军军医大学 | Tetanus toxin detection kit |
| CN112326972A (en) * | 2020-10-30 | 2021-02-05 | 安徽理工大学 | Chemiluminescence quantitative detection kit for detecting complete PINP in serum |
| CN113933497A (en) * | 2021-11-03 | 2022-01-14 | 辽宁中医药大学附属医院 | I type collagen amino-terminal extension peptide magnetic particle chemiluminescence immunoassay kit and detection method thereof |
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| CN109061176A (en) * | 2018-06-29 | 2018-12-21 | 江南大学 | A kind of total Type I collagen aminoterminal of detection serum extends the kit and application method of peptide |
| CN110317271A (en) * | 2019-07-02 | 2019-10-11 | 武汉云克隆科技股份有限公司 | Weight/the light chain variable region and encoding gene and recombinant antibodies of a kind of PINP recombinant antibodies |
| CN111007269A (en) * | 2019-11-15 | 2020-04-14 | 迪瑞医疗科技股份有限公司 | Total I type collagen amino-terminal extension peptide chemiluminescence immunoassay kit and preparation method thereof |
| CN111239384A (en) * | 2020-01-15 | 2020-06-05 | 中国人民解放军陆军军医大学 | Tetanus toxin detection kit |
| CN112326972A (en) * | 2020-10-30 | 2021-02-05 | 安徽理工大学 | Chemiluminescence quantitative detection kit for detecting complete PINP in serum |
| CN113933497A (en) * | 2021-11-03 | 2022-01-14 | 辽宁中医药大学附属医院 | I type collagen amino-terminal extension peptide magnetic particle chemiluminescence immunoassay kit and detection method thereof |
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