CN106918708A - A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin - Google Patents
A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin Download PDFInfo
- Publication number
- CN106918708A CN106918708A CN201511001059.3A CN201511001059A CN106918708A CN 106918708 A CN106918708 A CN 106918708A CN 201511001059 A CN201511001059 A CN 201511001059A CN 106918708 A CN106918708 A CN 106918708A
- Authority
- CN
- China
- Prior art keywords
- insulin
- reagent
- latex particle
- coated
- concentration
- 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
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- 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
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/537—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody
- G01N33/538—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody by sorbent column, particles or resin strip, i.e. sorbent materials
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
The application is related to a kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin.More specifically, the kit of the disclosure includes reagent 1 and reagent 2, and insulin monoclonal antibody is wherein contained in reagent 1, and the latex particle for being coated with insulin is contained in reagent 2.Monoclonal antibody is combined with insulin in reagent 2, and multiple conjugates are brought together, turbidity increase.When sample to be tested is added, the insulin in sample forms competition with the insulin in reagent 2, causes haze reduction.The content of insulin in sample to be tested is detected by the degree for reducing.The kit sensitivity of the disclosure is high, can detect serum or plasma sample, it is adaptable to which Biochemical Analyzer is easy to the diagnosis of diabetes to improve detection speed.
Description
Technical field
This disclosure relates to clinical examination field；It is more particularly to a kind of competition law latex enhancing immune for detecting insulin saturating
Penetrate than turbid kit.
Background technology
Insulin is a kind of hormone of islet β cell, is made up of 51 amino acid, and molecular weight is about 6000 dongles
, it is topmost hypoglycemic hormone in human body.Insulin is made up of (see Fig. 1) two peptide chains of A, B.Wherein A7 (Cys)-B7
(Cys), the sulfydryl in A20 (Cys)-B19 (Cys) four cysteines forms two disulfide bond, couples together the chain of A, B two.
In addition a disulfide bond is there is also in A chains between A6 (Cys) and A11 (Cys).
Insulin plays important influence power to the metabolic process of the various tissues of body, is to promote body anabolic
Important hormone, plays an important role on the metabolism of energy substances such as regulation body sugar, fat, protein：1) sugar is arranged
Preserve and use：When blood sugar concentration is raised, insulin secretion increases, and accelerates the sugar of blood to enter the tissue such as liver, muscle,
And preserved in the form of glycogen.When blood sugar level declines, insulin secretion is reduced, and the glycogen of storage can be made to come back to blood
For body provides energy in liquid.2) synthesis and storage of fat are helped：Insulin can promote liver synthetic fatty acid, make trigalloyl
Glycerine synthesis increases, and VLDL synthesis speeds.It can also suppress the activity of lipolytic enzyme, so as to suppress dividing for fat
Solution.3) synthesis of protein is helped：Insulin can promote amino acid to enter in histocyte, and synthesizing protein increases.Pancreas
Island element may also suppress the decomposition of protein, and the amino acid for making histocyte be released into blood is reduced.
Blood insulin inspection can be determined that diabetic is 1 type patient or 2 type patients.It is primarily suitable for not making
With the patient of insulin therapy, can be measured in empty stomach and blood drawing in 2 hours after the meal, Fasting insulin level should under normal circumstances
Should be 5-30 μ IU/ml, and level should be higher by 4-5 times than empty stomach after the meal.If the insulin level of patient is substantially reduced, just claim
For definitely shortage, it is seen that in type 1 diabetes；If do not significantly reduced, and blood glucose rise is shown as, be known as lacking relatively
It is weary, because the link that insulin plays a role breaks down, it is common in the diabetes B patient that there is insulin resistance.
The method of current clinical labororatory's detection insulin uses immunization method, and conventional method has radiommunoassay
Method, enzyme-linked immunosorbent assay, luminescent immunoassay.
Immunological method be using polypeptide drugs antigenic determinant position monoclonal or polyclonal antibody specifically
Identification checking matter, then quantified with radiating counting, the method such as colorimetric, will special antigen-antibody reaction be equipped with Sensitive Detection
Method.
A) measuring principle of radio immunoassay (Immunoradiometrec assay, IRMA) is：Measured object first with
Antibody complex formation in fixing phase, then with mark (125I) antibody is combined, and forms sandwich complex.Due to recognizing twice, this
The specificity of method is just considerably increased, is that a kind of sensitivity is high and the low assay method that makes a variation.It is disadvantageous in that to mark
The purity requirement of antibody is very high, while there is radiocontamination；
B) principle and IRMA of enzyme-linked immunosorbent assay (Enzyme-linked immunosorbent assay, ELISA)
Similar, simply second antibody is not to use labelled with radioisotope, but with can with substrate occur chromogenic reaction enzyme come
Mark, the proportionate relationship according to enzymic catalytic reaction product and the area of a room of insulin is come quantitative insulin.Compared with above-mentioned two method,
ELISA has long service life, reproducible, the advantage in radiationless source.But this kind of method used time be more long, the degree of accuracy and repetition
Property is not good enough；
C) luminescent immunoassay (Luminescent immunoassay, LIA) generally can be divided into fluorescence, phosphorescence and change
Learn luminous three kinds.It is coupled as tracer and antibody or antigen by the use of light emitting molecule, the luminous intensity according to light emitting molecule is determined
Tested insulin content.It is disadvantageous in that：Typically light unstable, to ask that disconnected, glitter lights, and in course of reaction
In easily fission, cause reaction result unstable；In addition.Need to be separated to combining phase, free phase during detection, operating procedure
Many, testing cost is high.
Many researchs show different detection methods or detecting system at aspects such as detection sensitivity, specificity and stability
There is significant difference.Clinically diagnosed or treated using such insulin assay result, it will brought deviation.Cause pancreas
The reason for island element testing result difference is many, including in immunological technique difference, normative reference reagent difference, clinical samples
The interference (especially IAA etc.) of Multiple components and the special sex chromosome mosaicism of other samples.It is insulin analog, anti-
Insulin antibody and heterophile antibody etc. differ greatly from the cross reaction of insulin detection antibody in different analysis methods.
Latex intensified transmission immunological turbidimetry detection (PETIA) technology is to develop to set up on the basis of latex agglutination qualitative test
A kind of on-radiation Advances in Homogeneous Immunoassay.The method first combines antibody (or antigen) and latex particle, when antigen is (or anti-
Body) when being reacted with the latex particle for combining antibody (or antigen), form Ag-Ab-latex particle (or antibody-antigene-
Latex particle) compound, so as to produce turbidity, calculate the content of antigen (or antibody) in sample.Carried out using Biochemical Analyzer
Turbidimetric assay so that whole analysis process only needs a few minutes.Compared with above-mentioned three kinds of detection methods, PETIA has taken into account higher
The advantage of the aspect such as sensitivity, repeatability, correlation, detection speed be fast, is adapted to Emergency call and detects, clinic popularization and application.
The content of the invention
Therefore, according to some implementation methods of the disclosure, there is provided a kind of Insulin Kit that can be detected in blood sample,
In particular to a kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin.The kit is adapted to detect for
Blood sample, including serum, blood plasma and whole blood.
In some embodiments, there is provided a kind of competition law turbid reagent of latex enhancing immune transmittance for detecting insulin
Box, it includes reagent 1 and reagent 2, and wherein reagent 1 contains insulin monoclonal antibody, the poly- agent of rush, buffer solution；Reagent 2 contains bag
There are latex particle, polymer, the buffer solution of insulin.
In some implementation methods of the disclosure, insulin monoclonal antibody is selected from：Rabbit-anti people, chicken are anti-human, the anti-human list of mouse
Clonal antibody.In a detailed embodiment, insulin monoclonal antibody is mouse anti-human monoclonal's antibody.The reagent of the disclosure
Box is not limited to the strain of specific monoclonal, any appropriate commercially available insulin monoclonal antibody or by approach well known
The monoclonal antibody of preparation is used equally to the disclosure.In some embodiments, insulin monoclonal antibody concentration is in reagent 1
0.01-0.2mg/ml, preferably 0.05-0.15mg/ml, more preferably 0.08-0.15mg/ml.In a detailed embodiment, try
Insulin monoclonal antibody concentration is 0.08mg/ml in agent 1；In another embodiment, insulin Dan Ke in reagent 1
Grand AC is 0.15mg/ml.
In some embodiments, the insulin antigen being coated with latex particle in reagent 2 is selected from：Pork insulin, ox
Insulin, semi-synthetic insulin, biosynthetic insulin.In a detailed embodiment, it is biosynthetic insulin.It is biological
Insulin synthesis refer to by the artificial synthesized insulin of biotechnology.Any appropriate commercially available biosynthetic insulin can use
In the disclosure.
In some embodiments, the latex particle for insulin being coated with described in reagent 2 is to be made by the steps
's：
Containing 0.1 to 1mg/ml (preferably 0.5mg/ml) EDAC and 0.1 to 1mg/ml (preferably 0.5mg/ml) S-NHS's
In 10 to 100mM (preferably 30 to 50mM) HEPES buffer solution (pH 7.2), 1% latex particle (w/v) is activated into 10- in 37 DEG C
30 minutes (preferably 20 minutes),
To insulin is added in the solution of said mixture, make its concentration for 0.05-0.8mg/ml, preferably 0.25 to
0.5mg/ml,
In 37 DEG C oscillating reactions 1-4 hours, preferably 3 hours, insulin is coated with to latex particle,
Supernatant is removed in centrifugation, and results are coated with the latex particle of insulin.
With the resuspended latex for being coated with insulin of Tris buffer solutions (pH 6.0) of 10-100mM (preferably 30 to 50mM)
Grain, ice-bath ultrasonic 30 minutes.
In some embodiments, insulin is made to be coated with to latex particle using chemical crosslink technique.In some embodiment party
In formula, insulin is set to be coated with to latex particle using crosslinking agent EDC and S-NHS, and latex particle is the polyphenyl of carboxyl modified
Ethene particle.In some embodiments, the concentration range of latex particle is 0.2-1.0% (w/v) during coating.
In some embodiments, the particle size range of latex particle is 100-250nm.In a detailed embodiment,
The particle diameter of latex particle is 200nm.
In some embodiments, the polymer in reagent 2 be selected from PEG2000, PEG4000, PEG6000, TWEEN20,
TWEEN40 and combinations thereof.In some embodiments, the polymer concentration range in reagent 2 is 1.0-10% (w/v).One
In individual specific embodiment, polymer concentration range is 1.0-2.0%.
In some embodiments, the poly- agent of rush in reagent 1 be selected from PEG2000, PEG4000, PEG6000, TWEEN20,
TWEEN40 and combinations thereof；Concentration range is 1.0-10% (w/v), preferably 1.0-2% (w/v).
In some embodiments, the buffer solution in reagent 1 and reagent 2 can be with identical, it is also possible to different.In some implementations
In mode, the buffer solution in reagent 1 and reagent 2 is independently selected from MES, Tris, MOPS and combinations thereof.In a specific embodiment party
In formula, buffer solution is MES.In another embodiment, buffer solution is Tris.In some embodiments, buffer solution
Concentration is 0.10-0.3mol/L.In some embodiments, buffer concentration is 0.15-0.25mol/L.In some embodiment party
In formula, the pH scopes of reagent 1 are 5.0-8.0, preferably 7.0-7.5；The pH scopes of reagent 2 are 6.0-8.5, preferably 6.0-7.0.
In some embodiments, also contain preservative in reagent 1 or reagent 2, one kind selected from Sodium azide and PC300 or
It is various.Concentration of preservatives scope is in 0.02-0.5% (w/v), preferably 0.05 to 0.15%, 0.1- in some embodiments
0.2%.
In some embodiments, as needed, the kit of the disclosure may be fitted with calibration object.Technical staff manages
Solution, calibration object is generally used for reagent calibration.With the kits calibration object of the application, obtain readings and draw standard curve.
When sample is measured, it is possible to which the readings of sample is found into corresponding insulin content on this curve.Therefore, it is any commercially available
Appropriate individual event or multinomial calibration object can be used in the disclosure, it is also possible to voluntarily prepare calibration object.In some embodiments, school
Quasi- product contain the insulin of concentration known.At least only need two points to be assured that straight line in fact, technical staff know.
But in some embodiments, calibration object is preferably directed at least 5 different concentration.In a detailed embodiment, when
For detecting during serum, calibration object be serum matrix, and containing concentration be respectively 400uIU/ml, 150uIU/ml, 70uIU/ml,
The Sodium azide of the insulin of 20uIU/ml, 10uIU/ml, 0uIU/ml and (w/v) 0.01-0.5%.
In a specific embodiment, there is provided a kind of competition law turbid examination of latex enhancing immune transmittance for detecting insulin
Agent box, it includes reagent 1, reagent 2 and calibration object,
Reagent 1 is included：
0.08mg/ml monoclonal antibodies, it is humanized murine antibodies, and potency is more than 1:30000, with high-affinity,
Tris buffer solution 0.20mol/L pH=7.5,
Tween 40 1.5% (w/v),
NaN30.2% (w/v),
Reagent 2 is included：
Tris buffer solution 0.25mol/L pH=6.0,
Be coated with the latex particle 200nm of insulin, 0.45% (w/v), wherein insulin equivalent to 0.35mg/ml, its
It is biosynthetic insulin,
PEG4000 2.0% (w/v)
Calibration object：Serum matrix, concentration is respectively 400,150,70,20,10,0uIU/ml.
In another embodiment, there is provided a kind of competition law latex enhancing immune transmittance for detecting insulin is turbid
Kit, it includes reagent 1, reagent 2 and calibration object,
Reagent 1 is included：
MES buffer solution 0.15mol/L pH=7.0,
Monoclonal antibody 0.15mg/ml, it is humanized murine antibodies, and potency is more than 1:20000, with high-affinity,
PEG2000 1.0% (w/v),
NaN30.1% (w/v),
Reagent 2 is included：
MES buffer solution 0.25mol/L pH=6.0,
The latex particle 250nm, wherein 0.90% (w/v), the concentration of insulin for being coated with insulin are 0.55mg/ml,
It is biosynthetic insulin,
TWEEN20 2.0% (w/v)
Calibration object：Serum matrix, concentration is respectively 400,150,70,20,10,0uIU/ml.
In some embodiments, there is provided a kind of method for preparing the latex particle for being coated with insulin, including step：
1) 0.1 to 1mg/ml (preferably 0.5mg/ml) EDAC and 0.1 to 1mg/ml (preferably 0.5mg/ml) S-NHS is being contained
10 to 100mM HEPES buffer solutions in by latex particle in 37 DEG C activate 10 to 30 minutes,
2) to step 1) obtained by solution in add insulin, make its concentration for 0.05-0.8mg/ml,
3) in 37 DEG C of oscillating reactions preferably 3 hours 1 to 4 hour, insulin is made to be coated with to the latex particle,
4) supernatant is removed in centrifugation, and results are coated with the latex particle of insulin.
In some embodiments, described insulin monoclonal antibody is selected from：Rabbit-anti people, chicken are anti-human, the anti-human Dan Ke of mouse
Grand antibody；It is preferred that insulin monoclonal antibody is mouse anti-human monoclonal's antibody.In some embodiments, insulin is selected from：Pig
Insulin, bovine insulin, semi-synthetic insulin, biosynthetic insulin；It is preferred that biosynthetic insulin.In some implementation methods
In, described insulin monoclonal antibody potency is more than 1:30000.In some embodiments, the particle diameter of latex particle is
100nm to 250nm, preferably 200nm.In some embodiments, during coating reaction, the concentration range of latex particle is based on w/v
It is 0.2 to 1.0%, preferably 1%.
In some embodiments, there is provided a kind of latex particle for being coated with insulin, it is by disclosed method
Obtained by preparation.
In some embodiments, there is provided the disclosure be coated with the latex particle of insulin prepare detection reagent in
Purposes.In some embodiments, detection reagent is the turbid reagent of latex enhancing immune transmittance.In some embodiments, examine
Test agent is the reagent for detecting insulin.
Brief description of the drawings
Fig. 1：Insulin structure.
Fig. 2：The calibration curve of disclosure Insulin Kit 1.
Fig. 3：Disclosure Insulin Kit 1 is schemed to external the related of chemical luminescence reagent kit detection serum measurement result.
Abscissa is uIU/ml；Ordinate is absorbance.
Fig. 4：Disclosure Insulin Kit 1 is schemed to external the related of chemical luminescence reagent kit detection plasma assay results.
Abscissa is uIU/ml；Ordinate is absorbance.
Specific embodiment
The disclosure will be further illustrated by following non-limiting examples below, it is as well known to those skilled in the art, not
In the case of disclosure spirit, many modifications can be made to the disclosure, such modification also falls into the scope of the present disclosure.
Embodiment
Embodiment 1：It is coated with the preparation method of the latex particle of insulin
In the 50mM of the 10ml containing 0.5mg/ml EDAC (being purchased from Merck) and 0.5mg/ml S-NHS (being purchased from Merck)
By the latex particle of 1% concentration, (particle diameter 200nm or 250nm, purchased from JSR, surface carboxyl groups are repaiied in HEPES buffer solution (pH 7.2)
Decorations) activated 20 minutes in 37 DEG C,
To insulin (being purchased from Fitzgerald) is added in above-mentioned solution, its concentration is set to reach 0.25mg/ml,
At 37 DEG C, oscillating reactions 3 hours makes insulin be coated with to latex particle,
Supernatant is removed in centrifugation, and results are coated with the latex particle of insulin.
With the resuspended latex particle for being coated with insulin of Tris buffer solutions (pH 6.0) of 50mM, ice-bath ultrasonic is after 30 minutes
Store for future use.
Embodiment 2：The preparation of Insulin Kit 1
1. the preparation of reagent 1：
According to following composition reagent preparation 1：
Monoclonal antibody 0.08mg/ml, mouse is anti-human, potency>1:30000 (are purchased from：Fitzgerald)
Tris buffer solution 0.20mol/L pH=7.5,
Tween 40 1.5% (w/v),
NaN30.2% (w/v).
2. the preparation of reagent 2：
According to following composition reagent preparation 2：
Tris buffer solution 0.25mol/L, pH=6.0,
Latex particle 0.45% (w/v) prepared by embodiment 1
PEG4000 2.0% (w/v),
3. the preparation of calibration object：
Human serum matrix, addition insulin (be purchased from Fitzgerald) concentration not Wei 400,150,70,20,10,0uIU/
Ml, and 0.09% (w/v) Sodium azide.
The preparation of the Insulin Kit 2 of embodiment 3.
1. the preparation of reagent 1：
According to following composition reagent preparation 1：
Monoclonal antibody 0.15mg/ml, mouse is anti-human, potency>1:30000 (are purchased from：Fitzgerald)
MES buffer solution 0.15mol/L, pH=7.0,
PEG2000 1.0% (w/v),
NaN30.1% (w/v)；
2. the preparation of reagent 2：
According to following composition reagent preparation 2：
MES buffer solution 0.25mol/L, pH=6.0,
Latex particle 0.90% (w/v) prepared by embodiment 1
TWEEN 20 2.0% (w/v).
3. the preparation of calibration object：
Normal human serum matrix, addition insulin (be purchased from Fitzgerald) concentration not Wei 400,150,70,20,10,
0uIU/ml, and 0.09% (w/v) Sodium azide.
Test case
The drafting of the standard curve of test case 1.
Kit 1 uses Olympus AU400 calibration curves, as a result sees Fig. 2.
Table 1：Absorbance
| Concentration (uIU/ml) | Absorbance A |
| 400 | 685 |
| 150 | 1500 |
| 70 | 3125 |
| 20 | 5613 |
| 10 | 6872 |
| 0 | 7900 |
Test case 2：The sensitivity behaviour evaluation of the application kit 1
Instrument：Olympus AU400, Chemiluminescence Apparatus.
Table 2：Parameter setting
Contrast agents box：Abbott Laboratories' insulin detection reagent (chemoluminescence method)
Cleaning Principle：The measure of insulin is 2 sandwich immunoassays of a kind of direct chemiluminescence and double antibody
The assay method being combined.First body is referred to as labeled antibody, is formed by monoclonal AIA mark acridinium ester；The
Two antibody are referred to as immobilized antibody, are to be combined shape with paramagnetic particle in the way of covalent bond by monoclonal AIA
Into.Insulin in serum produces light quantum after there is immune response with corresponding antibodies, its light quantity subnumber number and serum
The concentration of middle insulin is directly proportional, and the concentration level of serum insulin can be tried to achieve through standard curve.
Detection process：Serum is taken in specimen cup, is put on Full-automatic chemiluminescence immunoassay analysis meter, adjust detection ginseng
Number：The μ l of 25 the first reagents of μ l+ of serum 50 are incubated 5min in 37 DEG C, plus the μ l of the second reagent 250 are incubated 2.5min in 37 DEG C, with distillation
Water is separated to reaction cup, aspirated, cleaned, respectively plus 300 μ l acid reagents and base reagent are anti-to carrying out in reaction system finally
Should light, read light quantity subnumber, further according to standard curve, calculate result.
It is prepared by standard curve：Using two-point potentionmetric, standard serum 0.0uIU/ml and 138.0uIU/ml are placed in instrument
On the scaling position of device, mensuration program is incorporated into, upper machine determines automatic printing and goes out standard curve.
Test case 3：Sensitivity evaluation
The suitable serum of concentration is diluted with deionized water, a certain concentration is limited close to sensitivity, detected daily twice,
Continue 10 days, record initial data, such as following table is calculated.
(1) the sensitivity behaviour evaluation of kit 1, the results are shown in Table 3.
Table 3：The sensitivity measured value of kit 1
| Number of days | Concentration one | Concentration two | Concentration three |
| 1 | 1.63 | 2.27 | 3.61 |
| 2 | 1.19 | 2.38 | 3.49 |
| 3 | 1.42 | 2.11 | 3.68 |
| 4 | 1.29 | 2.24 | 3.29 |
| 5 | 1.33 | 2.41 | 3.56 |
| 6 | 1.54 | 2.33 | 3.62 |
| 7 | 1.46 | 2.24 | 3.49 |
| 8 | 1.39 | 2.01 | 3.67 |
| 9 | 1.55 | 2.15 | 3.46 |
| 10 | 1.22 | 2.03 | 3.34 |
| Average | 1.40 | 2.22 | 3.52 |
| SD | 0.15 | 0.14 | 0.13 |
| CV (%) | 10.43 | 6.28 | 3.77 |
(2) contrast agents box sensitivity evaluation, the results are shown in Table 4.
Table 4：Contrast agents box sensitivity measured value
| Number of days | Concentration one | Concentration two | Concentration three |
| 1 | 1.02 | 2.09 | 3.72 |
| 2 | 1.21 | 1.91 | 3.38 |
| 3 | 1.29 | 1.89 | 3.67 |
| 4 | 1.01 | 2.23 | 3.41 |
| 5 | 0.95 | 2.01 | 3.39 |
| 6 | 1.54 | 2.11 | 3.64 |
| 7 | 1.69 | 2.31 | 3.51 |
| 8 | 0.93 | 2.35 | 3.38 |
| 9 | 1.55 | 2.46 | 3.69 |
| 10 | 1.34 | 2.03 | 3.26 |
| Average | 1.25 | 2.14 | 3.51 |
| SD | 0.28 | 0.19 | 0.16 |
| CV (%) | 21.97 | 8.97 | 4.65 |
Evaluation criterion：In the daytime CV is less than 20%
Conclusion：The sensitivity of the detection kit 1 of the detection insulin of the disclosure is 1.40uIU/ml, contrast agents box
Sensitivity is 2.14uIU/ml, the sensitivity of the sensitivity better than contrast agents box of disclosure kit 1.
The correlation (serum sample) of the application kit 1 of test case 4.
Reagent, calibration object, parameter are ibid；Contrast agents are Abbott Laboratories' insulin detection reagent (chemoluminescence method)；Sample
It is serum.
Table 5：Detection data
Data to being obtained carry out correlation mapping, as a result see Fig. 3.Contrast agents detect the blood of below 2.14uIU/ml
This when of final proof, is it sometimes appear that negative value；Self-made reagent is more than 0.99 with the coefficient correlation of import reagent, with good correlation.
Test case 5：The correlation (plasma sample) of the application kit 1
Reagent, calibration object, parameter are ibid；Contrast agents are Abbott Laboratories' insulin detection reagent (chemoluminescence method)；Sample
It is blood plasma.
Table 6：Detection data
| Sample number | Kit 1 | Contrast agents | Sample number | Kit 1 | Contrast agents |
| 1 | 31 | 28 | 21 | 33 | 29 |
| 2 | 17 | 16 | 22 | 22 | 26 |
| 3 | 2.33 | 1.56 | 23 | 26 | 24 |
| 4 | 124 | 120 | 24 | 16 | 14 |
| 5 | 112 | 121 | 25 | 28 | 31 |
| 6 | 24 | 22 | 26 | 7.31 | 5.97 |
| 7 | 17 | 15 | 27 | 87 | 79 |
| 8 | 6.58 | 6.79 | 28 | 54 | 51 |
| 9 | 24 | 21 | 29 | 19 | 22 |
| 10 | 31 | 34 | 30 | 10 | 11 |
| 11 | 5 | 5 | 31 | 5.57 | 5.01 |
| 12 | 2 | 1.51 | 32 | 54 | 51 |
| 13 | 38 | 36 | 33 | 21 | 25 |
| 14 | 67 | 65 | 34 | 2.58 | 1.25 |
| 15 | 33 | 32 | 35 | 14 | 13 |
| 16 | 3.25 | 1.89 | 36 | 21 | 22 |
| 17 | 56 | 55 | 37 | 2.36 | 1.25 |
| 18 | 24 | 23 | 38 | 1.58 | -0.25 |
| 19 | 96 | 97 | 39 | 8.52 | 8.12 |
| 20 | 28 | 25 | 40 | 9.25 | 9.25 |
Data to being obtained carry out correlation mapping, as a result see Fig. 4.Contrast agents detect the blood of below 2.14uIU/ml
It sometimes appear that negative value during slurry sample；Self-made reagent is more than 0.99 with the coefficient correlation of import reagent, with good correlation.
Test case 6：The component and its test result of the application Insulin Kit 2
Kit 2 is detected according to the method for above-mentioned test case 3-5, as a result shows it in sensitivity, correlation
Energy aspect is similar with kit 1.
Table 7：The sensitivity measured value of kit 2
| Number of days | Concentration one | Concentration two | Concentration three |
| 1 | 1.23 | 2.35 | 3.65 |
| 2 | 1.29 | 2.28 | 3.58 |
| 3 | 1.22 | 2.24 | 3.68 |
| 4 | 1.18 | 2.25 | 3.44 |
| 5 | 1.28 | 2.44 | 3.67 |
| 6 | 1.44 | 2.18 | 3.49 |
| 7 | 1.52 | 2.25 | 3.45 |
| 8 | 1.29 | 2.18 | 3.47 |
| 9 | 1.45 | 2.06 | 3.58 |
| 10 | 1.32 | 2.07 | 3.38 |
| Average | 1.32 | 2.23 | 3.54 |
| SD | 0.11 | 0.12 | 0.11 |
| CV (%) | 8.45 | 5.21 | 3.02 |
Table 8：Virus monitory data
| Sample number | Kit 2 | Contrast agents | Sample number | Kit 2 | Contrast agents |
| 1 | 28 | 27 | 21 | 28 | 29 |
| 2 | 17 | 16 | 22 | 23 | 24 |
| 3 | 14 | 15 | 23 | 28 | 28 |
| 4 | 25 | 24 | 24 | 15 | 16 |
| 5 | 38 | 26 | 25 | 30 | 30 |
| 6 | 6 | 5 | 26 | 6.58 | 6.54 |
| 7 | 93 | 92 | 27 | 2.05 | 1.01 |
| 8 | 107 | 94 | 28 | 128 | 120 |
| 9 | 65 | 60 | 29 | 117 | 123 |
| 10 | 100 | 102 | 30 | 22 | 25 |
| 11 | 3 | 1.51 | 31 | 18 | 18 |
| 12 | 22 | 24 | 32 | 6.59 | 6.55 |
| 13 | 51 | 54 | 33 | 4.02 | 4 |
| 14 | 78 | 79 | 34 | 3.55 | 2.22 |
| 15 | 254 | 249 | 35 | 3.07 | 1.89 |
| 16 | 365 | 334 | 36 | 1.07 | -0.09 |
| 17 | 4.17 | 4.16 | 37 | 2.04 | -0.02 |
| 18 | 13 | 12 | 38 | 1.05 | -1.23 |
| 19 | 27 | 27 | 39 | 1.08 | -0.07 |
| 20 | 30 | 30 | 40 | 0.58 | -0.28 |
Table 9：Blood plasma detection data
| Sample number | Kit 2 | Contrast agents | Sample number | Kit 2 | Contrast agents |
| 1 | 30 | 28 | 21 | 32 | 29 |
| 2 | 18 | 16 | 22 | 25 | 26 |
| 3 | 2.45 | 1.56 | 23 | 28 | 24 |
| 4 | 118 | 120 | 24 | 14 | 14 |
| 5 | 118 | 121 | 25 | 30 | 31 |
| 6 | 22 | 22 | 26 | 6.58 | 5.97 |
| 7 | 15 | 15 | 27 | 78 | 79 |
| 8 | 6.66 | 6.79 | 28 | 52 | 51 |
| 9 | 22 | 21 | 29 | 20 | 22 |
| 10 | 35 | 34 | 30 | 9 | 11 |
| 11 | 6 | 5 | 31 | 5.65 | 5.01 |
| 12 | 2.05 | 1.51 | 32 | 52 | 51 |
| 13 | 35 | 36 | 33 | 22 | 25 |
| 14 | 68 | 65 | 34 | 2.68 | 1.25 |
| 15 | 31 | 32 | 35 | 13 | 13 |
| 16 | 3.58 | 1.89 | 36 | 22 | 22 |
| 17 | 54 | 55 | 37 | 2.58 | 1.25 |
| 18 | 23 | 23 | 38 | 1.44 | -0.25 |
| 19 | 95 | 97 | 39 | 8.68 | 8.12 |
| 20 | 25 | 25 | 40 | 9.14 | 9.25 |
The disclosure uses competitive immunoreaction, due to making monoclonal antibody using latex intensified method and having been integrated into latex
The conjugate volume that insulin response on particle is formed is larger, and absorbance being capable of significant change at a particular wavelength.Serum or
The presence of insulin forms competition in blood plasma, causes the insulin for being coated in latex particle that reduction is combined with monoclonal antibody, special
Standing wave lower haze reduction long, the slight change of insulin to be measured leads to the significant change of absorbance.So this kit
Another distinguishing feature is sensitivity high, can sensitively reflect the content of test serum or plasma insulin.
In the market has the kit of panimmunity transmittance purifying method, but may not apply to Biochemical Analyzer, during detection
Between it is relatively long, influence testing result promptness.The disclosure uses competition law Latex-enhanced immunoturbidimetric assay, it is adaptable to
Biochemical Analyzer.So a feature of this kit is to shorten detection time, detection speed is improved.
Claims (14)
1. a kind of turbid kit of latex enhancing immune transmittance for detecting insulin, it includes reagent 1 and reagent 2,
Wherein：
Reagent 1 contains：Insulin monoclonal antibody, the poly- agent of rush, buffer solution；
Reagent 2 contains：It is coated with latex particle, polymer, the buffer solution of insulin.
2. kit according to claim 1, wherein：
The latex particle for being coated with insulin is made by the steps：
Latex particle is activated 10-30 minutes in the 10-100mM HEPES buffer solutions containing crosslinking agent,
Insulin is added, makes its concentration for 0.05-0.8mg/ml,
Oscillating reactions 1-4 hours, preferably 3 hours, insulin is set to be coated with to latex particle,
Supernatant is removed in centrifugation, and results are coated with the latex particle of insulin；
It is preferred that, the concentration range of latex particle is calculated as 0.2 to 1.0% by w/v during coating；
The particle diameter of latex particle is 100 to 250nm, preferably 200nm.
3. kit according to claim 1, wherein：
Insulin monoclonal antibody concentration is 0.01-0.2mg/ml, more preferably preferably 0.05-0.15mg/ml, 0.08- in reagent 1
0.15mg/ml；
Insulin monoclonal antibody is selected from：Rabbit-anti people, chicken are anti-human, mouse anti-human monoclonal's antibody, and insulin monoclonal antibody is preferred
It is mouse anti-human monoclonal's antibody；
The insulin being coated with latex particle is selected from：Pork insulin, bovine insulin, semi-synthetic insulin, biosynthesis pancreas islet
Element；It is preferred that biosynthetic insulin.
4. kit according to claim 1, wherein the poly- agent of described rush be selected from PEG2000, PEG4000, PEG6000,
TWEEN 20, TWEEN 40 and combinations thereof, concentration is calculated as 0.5 to 5%, preferably 1 to 2% by w/v.
5. kit according to claim 2, wherein：
Insulin is set to be coated with to latex particle using chemical crosslink technique；Crosslinking agent be 0.1 to 1mg/ml EDC and 0.1 to
The S-NHS of 1mg/ml；Latex particle is the granules of polystyrene of carboxyl modified.
6. kit according to claim 1, wherein：
Polymer is selected from PEG2000, PEG4000, PEG6000, TWEEN20 and TWEEN40 and combinations thereof, and preferred concentration range is pressed
W/v is calculated as 1.0-10%.
7. kit according to claim 1, wherein：
Buffer solution in reagent 1 or reagent 2 is each independently selected from MES, Tris, MOPS and combinations thereof,
The pH scopes of reagent 1 are 5.0-8.0, preferably 7.0-7.5；
The pH scopes of reagent 2 are 6.0-8.5, preferably 6.0-7.0；
Preferred buffer concentration is 0.10-0.3mol/L, more preferably 0.15-0.25mol/L.
8. kit according to claim 1, wherein：
Also contain preservative in reagent 1 or reagent 2, described preservative is selected from Sodium azide, PC300 and combinations thereof, preferred concentration
0.02-0.5% is calculated as by w/v.
9. kit according to claim 1, it also includes calibration object, and described calibration object contains the pancreas islet of concentration known
Element, preferably at least 5 insulin of various concentrations；
It is further preferred that calibration object is respectively 400uIU/ml, 150uIU/ml, 70uIU/ml, 20uIU/ comprising serum matrix, concentration
The Sodium azide of the insulin of ml, 10uIU/ml, 0uIU/ml and the 0.01-0.5% based on w/v.
10. a kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin, it contains：
Reagent 1, it contains：
Reagent 2, it contains：
Tris buffer solution 0.25mol/L, pH=6.0,
The latex particle 200nm of insulin, based on w/v 0.45%, wherein insulin 0.35mg/ml are coated with,
PEG4000 based on w/v 2.0%；
Calibration object, it contains：
Serum matrix and concentration are respectively 400,150,70,20,10, the insulin of 0uIU/ml；
Wherein, the described latex particle for being coated with insulin is made by the steps：
1) latex particle is activated 10-30 minutes in the 10-100mM HEPES buffer solutions containing crosslinking agent,
2) insulin is added, makes its concentration for 0.05-0.8mg/ml,
3) oscillating reactions 1-4 hours, preferably 3 hours, insulin is made to be coated with to latex particle,
4) supernatant is removed in centrifugation, and results are coated with the latex particle of insulin；
The insulin monoclonal antibody is humanized murine antibodies, and potency is more than 1:30000；
The insulin is biosynthetic insulin.
A kind of 11. competition law turbid kits of latex enhancing immune transmittance for detecting insulin, it contains：
Reagent 1, it contains：
Reagent 2, it contains：
MES buffer solution 0.25mol/L, pH=6.0,
The latex particle 250nm based on w/v 0.90% of insulin is coated with, the wherein concentration of insulin is 0.55mg/ml,
TWEEN 20 based on w/v 2.0%；
Calibration object, it contains：
Serum matrix, insulin concentration is respectively 400,150,70,20,10,0uIU/ml；
Wherein, the latex particle for being coated with insulin is made by the steps：
1) latex particle is activated 10-30 minutes in the 10-100mM HEPES buffer solutions containing crosslinking agent,
2) insulin is added, makes its concentration for 0.05-0.8mg/ml,
3) oscillating reactions 1-4 hours, preferably 3 hours, insulin is made to be coated with to latex particle,
4) supernatant is removed in centrifugation, and results are coated with the latex particle of insulin；
The insulin monoclonal antibody is humanized murine antibodies, and potency is more than 1:30000；
The insulin is biosynthetic insulin.
A kind of 12. preparation methods of the latex particle for being coated with insulin, it includes step：
1) by latex in 10 to the 100mM HEPES buffer solutions containing 0.1 to 1mg/ml EDC and 0.1 to 1mg/ml S-NHS
Particle is activated 10 to 30 minutes,
2) to step 1) obtained by solution in add insulin, make its concentration for 0.05-0.8mg/ml,
3) oscillating reactions preferably 3 hours 1 to 4 hour, make insulin be coated with to the latex particle,
4) supernatant is removed in centrifugation, and results are coated with the latex particle of insulin,
Wherein described insulin monoclonal antibody is selected from：Rabbit-anti people, chicken are anti-human, mouse anti-human monoclonal's antibody；It is preferred that insulin
Monoclonal antibody is mouse anti-human monoclonal's antibody；
Insulin is selected from：Pork insulin, bovine insulin, semi-synthetic insulin, biosynthetic insulin；It is preferred that biosynthesis pancreas islet
Element；
Described insulin monoclonal antibody potency is more than 1:30000；
The particle diameter of latex particle be 100nm to 250nm, preferably 200nm,
The concentration range of latex particle is calculated as 0.2 to 1.0% by w/v during coating.
A kind of 13. latex particles for being coated with insulin, it is as obtained by prepared by the method described in claim 12.
Purposes of the latex particle of insulin in detection reagent is prepared is coated with described in 14. claims 13.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511001059.3A CN106918708A (en) | 2015-12-28 | 2015-12-28 | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511001059.3A CN106918708A (en) | 2015-12-28 | 2015-12-28 | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106918708A true CN106918708A (en) | 2017-07-04 |
Family
ID=59455059
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201511001059.3A Pending CN106918708A (en) | 2015-12-28 | 2015-12-28 | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106918708A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107741493A (en) * | 2017-09-30 | 2018-02-27 | 湖南海源医疗科技股份有限公司 | A kind of kit using Immune competition turbidimetry for Determination microdose urine protein |
| CN108362895A (en) * | 2018-02-12 | 2018-08-03 | 北京九强生物技术股份有限公司 | Folic acid detection kit and preparation method thereof |
| CN108776231A (en) * | 2018-09-06 | 2018-11-09 | 长沙文瀚生物技术有限责任公司 | A kind of Alb in human urine latex intensified secondary antibody competition immunoturbidimetry detection kit and its making and use method |
| CN109813908A (en) * | 2018-12-29 | 2019-05-28 | 宁波普瑞柏生物技术股份有限公司 | Eliminate the method and kit of hook effect in cystatin C detection |
| CN109975550A (en) * | 2018-12-29 | 2019-07-05 | 宁波普瑞柏生物技术股份有限公司 | Eliminate the method and kit of hook effect in β2-microglobulin detection |
| CN112946254A (en) * | 2021-01-18 | 2021-06-11 | 上海云泽生物科技有限公司 | Latex-enhanced competitive immunoturbidimetric assay method and kit |
| CN112969922A (en) * | 2018-11-09 | 2021-06-15 | 积水医疗株式会社 | Method for preventing abnormal detection in immunoassay of automatic analyzer, and immunoassay reagent |
| CN114047338A (en) * | 2021-11-10 | 2022-02-15 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102246039A (en) * | 2009-07-21 | 2011-11-16 | 积水医疗株式会社 | Insulin measurement method |
| CN102628868A (en) * | 2011-12-30 | 2012-08-08 | 北京九强生物技术股份有限公司 | Latex enhanced immunoturbidimetry kit for detection of asymmetric dimethylarginine content |
| CN102680712A (en) * | 2012-06-18 | 2012-09-19 | 王钊 | Competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and preparation method thereof |
| CN103328509A (en) * | 2009-12-25 | 2013-09-25 | 积水医疗株式会社 | Method for measuring human insulin and measurement reagent |
| CN104237498A (en) * | 2013-06-13 | 2014-12-24 | 绍兴圣康生物科技有限公司 | Preparation method of insulin determination kit employing latex enhanced turbidimetric immunoassay |
-
2015
- 2015-12-28 CN CN201511001059.3A patent/CN106918708A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102246039A (en) * | 2009-07-21 | 2011-11-16 | 积水医疗株式会社 | Insulin measurement method |
| CN103328509A (en) * | 2009-12-25 | 2013-09-25 | 积水医疗株式会社 | Method for measuring human insulin and measurement reagent |
| CN102628868A (en) * | 2011-12-30 | 2012-08-08 | 北京九强生物技术股份有限公司 | Latex enhanced immunoturbidimetry kit for detection of asymmetric dimethylarginine content |
| CN102680712A (en) * | 2012-06-18 | 2012-09-19 | 王钊 | Competitive latex-particle-enhanced immunoturbidimetric assay kit for BNP (B-type natriuretic peptide) and preparation method thereof |
| CN104237498A (en) * | 2013-06-13 | 2014-12-24 | 绍兴圣康生物科技有限公司 | Preparation method of insulin determination kit employing latex enhanced turbidimetric immunoassay |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107741493A (en) * | 2017-09-30 | 2018-02-27 | 湖南海源医疗科技股份有限公司 | A kind of kit using Immune competition turbidimetry for Determination microdose urine protein |
| CN108362895A (en) * | 2018-02-12 | 2018-08-03 | 北京九强生物技术股份有限公司 | Folic acid detection kit and preparation method thereof |
| CN108362895B (en) * | 2018-02-12 | 2020-05-08 | 北京九强生物技术股份有限公司 | Folic acid detection kit and preparation method thereof |
| CN108776231A (en) * | 2018-09-06 | 2018-11-09 | 长沙文瀚生物技术有限责任公司 | A kind of Alb in human urine latex intensified secondary antibody competition immunoturbidimetry detection kit and its making and use method |
| CN112969922A (en) * | 2018-11-09 | 2021-06-15 | 积水医疗株式会社 | Method for preventing abnormal detection in immunoassay of automatic analyzer, and immunoassay reagent |
| CN109813908A (en) * | 2018-12-29 | 2019-05-28 | 宁波普瑞柏生物技术股份有限公司 | Eliminate the method and kit of hook effect in cystatin C detection |
| CN109975550A (en) * | 2018-12-29 | 2019-07-05 | 宁波普瑞柏生物技术股份有限公司 | Eliminate the method and kit of hook effect in β2-microglobulin detection |
| CN112946254A (en) * | 2021-01-18 | 2021-06-11 | 上海云泽生物科技有限公司 | Latex-enhanced competitive immunoturbidimetric assay method and kit |
| CN112946254B (en) * | 2021-01-18 | 2024-03-15 | 上海云泽生物科技有限公司 | Latex-enhanced competitive immune turbidimetry detection method and kit |
| CN114047338A (en) * | 2021-11-10 | 2022-02-15 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
| CN114047338B (en) * | 2021-11-10 | 2024-03-12 | 上海捷门生物技术有限公司 | Urine transferrin detection kit and detection method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106918708A (en) | A kind of competition law turbid kit of latex enhancing immune transmittance for detecting insulin | |
| US11959912B2 (en) | Fluorescence immunochromatographic detection card and a preparation method therefor and use thereof | |
| CN102759631B (en) | The latex enhancing immune of a kind of quantitative detection Procalcitonin PCT is than turbid kit | |
| US10472400B2 (en) | Cardiac troponin I ultra-sensitive detection reagent kit, and ultra-sensitive detection method therefor | |
| CN102798725B (en) | Diagnostic kit for determination of serum total IgE, preparation method and application method | |
| CA2348379C (en) | Immunoassay for measuring human c-peptide and kit therefor | |
| DK164944B (en) | PROCEDURE, FITTING AND ANALYSIS OF DETERMINING CONCENTRATIONS OF MULTIPLE ANALYTES IN A FLUID | |
| CN102662061A (en) | Kit for determination of human alpha-fetoprotein content by latex-enhanced immunoturbidimetry | |
| CN108445230B (en) | Procalcitonin chemiluminescence detection reagent based on nano antibody and detection method | |
| CN107543932A (en) | The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of calcitonin | |
| CN103018464A (en) | Reagent for determining procalcitonin and preparation method of reagent | |
| CN102901812A (en) | Magnetic particle chemiluminescence immunoassay kit and assay method for human thyroglobulin antibodies (TGAb) | |
| CN109444437A (en) | Miao's Le Shi pipe hormone determination kit based on restoring method label attached luminescent indicator | |
| CN111239421A (en) | Latex-enhanced immunoturbidimetry kit for quantitatively detecting adiponectin ADPN and preparation and application methods thereof | |
| CN109001472A (en) | Human thyrotropin receptor antibody chemical luminescence detection kit and preparation method thereof and application method | |
| CN112698041A (en) | Compound, growth differentiation factor 15 detection kit thereof and application | |
| CN110790836A (en) | Monoclonal antibody of human inhibin B, application and kit thereof | |
| CN109187971A (en) | Neuronspecific enolase chemiluminescence immune detection reagent kit and preparation method thereof | |
| CN202916286U (en) | Latex enhanced turbidimetric immunoassay kit for quantitatively detecting procalcitonin (PCT) | |
| CN105067599A (en) | Chemiluminescence immune detection kit for detecting pro-gastrin-releasing peptide | |
| CN103969447A (en) | Vascular endothelial growth factor quantitative determination kit and light-emitting substrate solution thereof | |
| JP4197393B2 (en) | Test method for IgA nephropathy | |
| CN102818903A (en) | Double-particle compounded Lp-a detection kit | |
| TW201802472A (en) | Anti-human hemoglobin monoclonal antibody or antibody kit, insoluble carrier particle to which anti-human hemoglobin monoclonal antibody is immobilized, and measurement reagent and measurement method using same | |
| CN114113637A (en) | Thyroid stimulating hormone receptor antigen reagent and thyroid stimulating hormone receptor antibody quantitative determination kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170704 |