CN107102140A - A kind of glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent and preparation method thereof - Google Patents
A kind of glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent and preparation method thereof Download PDFInfo
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- CN107102140A CN107102140A CN201611018770.4A CN201611018770A CN107102140A CN 107102140 A CN107102140 A CN 107102140A CN 201611018770 A CN201611018770 A CN 201611018770A CN 107102140 A CN107102140 A CN 107102140A
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- glutamic acid
- preparation
- decarboxylase
- acridinium ester
- liquid
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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
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
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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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
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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
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/901—Antibodies with enzymatic activity; e.g. abzymes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2446/00—Magnetic particle immunoreagent carriers
- G01N2446/80—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2458/00—Labels used in chemical analysis of biological material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/04—Endocrine or metabolic disorders
- G01N2800/042—Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
Abstract
The invention discloses a kind of glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent, the kit includes:The coated magnetic particle working solution of glutamate decarboxylase, the glutamate decarboxylase working solution of the purifying of acridinium ester label, glutamic acid decarboxylase antibody calibration product, preexciting liquid, the exciting liquid of purifying.The invention also discloses a kind of preparation method of glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent in addition.Kit of the present invention is compared with available reagent box, with easy to operate, sensitivity height, the features such as detection range is wide.
Description
Technical field
It is specifically, anti-the invention provides a kind of glutamate decarboxylase the present invention relates to in-vitro diagnosis field of immunodetection
Body chemiluminescence immune detection reagent kit and preparation method thereof.
Background technology
Glutamate decarboxylase(GAD)It is synthesis gamma aminobutyric acid(GABA)Rate-limiting enzyme, with insulin in beta Cell of islet
Exist together presence, secretes simultaneously, the function with autocrine and the synthesis of paracrine Signal Regulation β cells and excreting insulin.Paddy ammonia
Acid decarboxylase antibody(GADA)In positive patient, on the one hand due to there is GADA in vivo, GADA have impact on after being combined with GAD
GABA aggregate velocity, disturbs the regulation of insulin synthesis and secretion, the insulin synthesis that pancreas islet β is carefully run is affected,
Cause the insufficient insulin in body-internal-circulation, so as to cause the diabetes that insulin is relied on.On the other hand, the GADA that body is produced
Excite T lymphocytes and cause the destruction of β cells.
Most type 1 diabetes are caused by immune-mediated islet p-cell destruction.This selective destruction with it is thin
Born of the same parents are immune relevant with humoral immunity.In addition to macrophage, lymphocytic infiltration, islet cell antibodies(ICA), glutamate decarboxylase
Antibody(GADA), insulin autoantibodies(IAA)The morbidity of type 1 diabetes can be forecast.In many autoantibodies, GADA goes out
Now the early, duration is long.The 3-5 after the diagnosis of 1 patients with type Ⅰ DM, ICA antibody titers and recall rate are remarkably decreased, 10 years
Only 20% patient is positive afterwards.On the contrary, GADA falls are small, 54% patient remains to detection after type 1 diabetes are diagnosed 10 years
GADA.Meanwhile, compared with ICA, GADA determines simple, and expense is low, it is easy to standardize.Therefore using the high GADA inspections of sensitivity
Survey, type 1 diabetes patient can be diagnosed to be from people at highest risk with as much as possible.
Latent autoimmune diabetes in adults(LADA)Early stage, clinical characters are much like with diabetes B, but short
Insulin therapy is dependent in the short several years, is essentially type 1 diabetes, under-weight index is shown as, ICA and some other
Autoantibodies.The amynologic index that GADA is diagnosed as LADA, is better than ICA.GADA, which is checked, is conducive to early detection
Patient LADA, strengthens follow-up, gives insulin therapy in early days, can protect the islet beta cell function of remaining, reduces in the recent period and remote
Phase complication.
Diabetes B be by insulin resistance and(Or)It is relative to lack what is caused, caused by not immune-mediated inflammation of pancreatic islet.
Therefore diabetes B patient GADA positive rates are less than type 1 diabetes patient, in diabetes B, initially i.e. using the disease of insulin
People and the negative patients of GADA, what its insulin β cell functions attenuation amplitude will be low is more, and the GADA of high titre imply that faster
Islet beta cell function exhaustion.Therefore, GADA state is to predict the ideal indicator of its clinical process during onset diabetes, and it can
So that LADA to be made a distinction with diabetes B.To sum up GADA detection is in the prediction of diabetes, and diagnosis and correct parting aspect are
With important value.
The common methods of clinical detection glutamic acid decarboxylase antibody have IIF, enzyme linked immunosorbent assay, but
These methods are all existed in place of some shortcomings.
First, IIF
The general principle of the method is to form antigen antibody complex with after the antigen binding in specific antibody and slide, after
Combined with fluorescence antibody with antigen antibody complex, form antigen-antibody fluorescent composition.Under fluorescence microscope, according to compound
The luminous situation of thing determines detected antibody.This method is evaluated:Because the fluorescence combined on antigen antibody complex resists
Body increases, and the fluorescent brightness sent is strong, thus its sensitiveness is strong.But its deficiency is also apparent:
(1) operate relative complex, it is necessary to which the fluorescence microscope of price costly, is difficult to promote, also not in many basic hospitals
It is applied to very much the more laboratory of specimen amount;
(2) background in fluoremetry is higher, can only carry out qualitative detection, immunofluorence technic, which is used for quantitative determination, to be had necessarily
It is difficult;
(3) result judgement needs experienced professional, and the objectivity of analysis result is not enough;
2nd, enzyme linked immunosorbent assay
Enzyme linked immunosorbent assay (ELISA) is widely used, but there is also following weak points for this method:
(1) 12 × 8 types, 6 × 8 types, 8 × 12 types or the hole Special micro porous plate of complete plate 96 is used to be used as antigen coat apparatus and anti-
Container is answered, 12 batches, 6 batches, 8 batches or whole plate first use can only be divided into when in use, it is impossible to independent, single part is carried out
Detection;
(2) reagent type used in quantitative determining is more, and each detection reagent will be contained with reagent bottle, and often be made
It is required for changing imbibition nozzle during with a kind of reagent being filled into the micropore of microwell plate respectively, not only reagent bottle species is more, filling
The operation of reagent is also extremely cumbersome;
(3) the corresponding mark to detection information is lacked, can only be by checking that the mark of kit external packing box just will appreciate that or know
The product batch number and term of validity information of detection reagent are known, and the information known is uncontrolled in detection process, with very big
Randomness;
(4) detection reagent easily causes the cross pollution between various reagents and shadow in open space in detection process
Ring the accuracy of testing result;
(5) use hand-manipulated detection process, the dosage of reagent or sample is not bery accurate, operating process is extremely cumbersome and multiple more
It is miscellaneous, easily occur bust, the degree of accuracy of testing result and precision are poor.
The content of the invention
Current glutamic acid decarboxylase antibody detection technique has the following disadvantages:Testing cost is high, detection sensitivity is low, detection
The range of linearity is narrow, reappearance is low, can not quantify, complex operation etc..
The present invention discloses that a kind of testing cost is low, sensitivity is high, detection is linear precisely in order to overcome shortcoming described above
Scope is wide, reappearance is high, can quantify, glutamic acid decarboxylase antibody kit simple to operate and preparation method thereof.The present invention
Chemical luminescence immune analysis reagent box is prepared first, is mainly included:The coated magnetic particle of glutamate decarboxylase, glutamate decarboxylase
Coated acridinium ester and glutamic acid decarboxylase antibody calibrate product;Then using Full-automatic chemiluminescence immunoassay analysis meter to calibration
Product are detected, draw standard curve, are built in computer software, test actual sample, and it is dense to calculate sample according to sample luminous value
Degree;Performance finally is carried out to glutamic acid decarboxylase antibody automatic chemiluminescence immunoassay system(Sensitivity, linear, precision
Degree, interference)Evaluation.
It is of the invention compared with current technology, with advantages below:
1st, present invention selection acridinium ester is as marker material, and applied to chemiluminescence immunoassay system, and the luminescence system is
Direct chemiluminescence, compared with traditional enzyme-catalyzed chemical luminescence, the reaction does not need the participation of enzyme, more cost-effective;
2nd, the acridinium ester chemiluminescent immunoassay system detection sensitivity that the present invention is selected is high, can reach 0.11 IU/mL,
10 times are at least improved compared to other glutamic acid decarboxylase antibody detection method sensitivity;
3rd, the acridinium ester chemiluminescent immunoassay system range of linearity that the present invention is selected is wide, can reach 10-1500 IU/mL, its
The inspection range of linearity of its glutamic acid decarboxylase antibody chemistry hair detection method is 20-1000 IU/mL;
4th, the acridinium ester chemiluminescent immunoassay system repeatability that the present invention is selected is high, criticizes interior and difference between batch within 5%,
This is that other chemiluminescence immunoassay systems are unapproachable;
5th, chemiluminescence immunoassay system of the invention has realized quantifying for sample, soft to testing by built-in standard curve
Part, only needs test sample to directly obtain the concentration value of sample;
6th, chemiluminescence immunoassay system of the invention has realized full-automation, and the addition of reagent and sample has instrument complete entirely
Into operation is easier, reduces artificial error.
Brief description of the drawings
Fig. 1 is the glutamic acid decarboxylase antibody canonical plotting that embodiment 3 is obtained.
Embodiment
Embodiment 1:Glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent preparation method
(1)It is prepared by the coated nanometer magnetic bead of glutamate decarboxylase:
Take the magnetic particle of 50mg carboxylated(Particle diameter is 0.05-1um)Suspension, Magneto separate removes supernatant, and it is 5.5 to use 0.02 M, pH
MES buffer solutions are resuspended, and add the EDC aqueous solution for 10 mg/mL that 0.5-2mL is newly configured, and activated magnetic beads surface carboxyl groups add 3-5
Mg glutamic acid decarboxylases, suspension 2-10 h, Magneto separate, remove supernatant at room temperature, are 8.0 with the 0.1 M pH containing 2% BSA
Tris buffer solutions are resuspended to 1mg/mL, obtain the coated magnetic particle of glutamic acid decarboxylase antibody monoclonal antibody, every bottle of 5mL packing
Be stored in 4 DEG C it is standby.
(2)The preparation of the acridinium ester of glutamate decarboxylase mark:
50 uL 2.5mg/mL glutamate decarboxylase is taken, the carbonate for adding 150 uL 0.1-0.2 M pH 9.0-9.5 delays
Fliud flushing, is mixed, and the acridinium ester for then adding the mg/mL of 1-2 uL 5 is mixed, and lucifuge is reacted at room temperature, is taken out after 1-2 h, with 2
Handled respectively with pure water and TBS buffer solutions first in mL zeba centrifugation desalting column desalting processings, desalination processes, finally
The acridine ester solution of obtained glutamate decarboxylase note is added, liquid to the preservation collected in centrifuge tube is in control glutamic acid decarboxylase
The acridinium ester of enzyme mark, every bottle of 5 mL packing be stored in 4 DEG C it is standby.
(3)Glutamic acid decarboxylase antibody calibrates the preparation of product:
Use standard items buffer solution(40 mM Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0)By glutamic acid decarboxylase antibody
Concentration is configured to for 5 IU/mL, 20 IU/mL, 100 IU/mL, 400 IU/mL, 800 IU/mL, 2000 IU/mL, every bottle
0.5 mL packing is lyophilized, and 4 DEG C save backup.
(4) preparation of chemiluminescence preexciting liquid:
1.0 liters of purified waters are measured, the hydrogen peroxide (H that 80uL mass fractions are 20% is sequentially added2O2), 1.0 grams of sodium azide, 1.5
Gram polysorbas20, shakes up rear lucifuge storage.
(5) preparation of chemiluminescence exciting liquid:
1.0 liters of purified waters are measured, 0.6 gram of sodium hydroxide, 0.5 gram of PC300,0.5g sodium azide, 1.5 grams of Triton are sequentially added
X-405, shakes up rear lucifuge storage.
Embodiment 2:Glutamic acid decarboxylase antibody chemical luminescence immunologic detection method:
The present invention is using Full-automatic chemiluminescence immunoassay analysis meter as detection instrument, and methodology pattern of the invention is double antigens sandwich
Method, i.e. instrument sequentially add 25 uL sample, 50 the uL coated magnetic particle of glutamate decarboxylase and 50 uL glutamic acid
After the coated acridinium ester of decarboxylase antibody, 10 min of reaction, Magneto separate is carried out, reactant mixture is sent into darkroom by instrument, successively
Add 50uL chemiluminescence preexcitings liquid, 50uL chemiluminescences exciting liquid and carry out luminescence-producing reaction, luminous intensity is finally recorded, from mark
Directrix curve calculates the glutamic acid decarboxylase antibody content of sample.
Embodiment 3:Glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent performance evaluation
Detection curve is shown in accompanying drawing 1.
The detection of sensitivity for analysis:
With reference to CLSI EP17-A file recommendation experimental programs, glutamic acid decarboxylase antibody chemical luminescence immunologic function test reagent is calculated
The sensitivity for analysis of box, the sensitivity for analysis tried to achieve is 0.11 IU/mL.
Linear detection:
It is 5 IU/mL, 20 IU/mL, 100 IU/mL, 400 IU/mL, 800 IU/mL, 2000 IU/mL standards to concentration
Product do linear analysis, calculate linearly dependent coefficient, r=0.9996, in addition, the kit is examined to glutamic acid decarboxylase antibody sample
The range of linearity of survey is 5-2000 IU/mL.
Precision is determined:
It is two glutamic acid decarboxylase antibody samples of 20 IU/mL and 400 IU/mL to take concentration, and each each concentration of sample is respectively done
3 parallel, is detected with three batches of kits, and calculating kit criticizes interior and difference between batch, as a result shows in the kit batch and criticizes
Between difference be respectively less than 5%.
Interference is tested:
Taking pooled serum to add chaff interference respectively includes:Bilirubin, hemoglobin, ascorbic acid, glyceride, adding proportion according to
1:20 are carried out, and are determined pooled serum respectively and be with the addition of the measured value of pooled serum after various chaff interferences, calculate therebetween
Deviation, with ± 10% for tolerance interval.As a result show, interference reaches NCCLS file standard, available for clinical real
Test room glutamic acid decarboxylase antibody accurate evaluation.
Embodiment 4:The sensitivity for analysis contrast experiment of glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent point
Concentration is not examined for 0 IU/mL Sample dilution with chemical luminescence detection method and traditional enzyme linked immunosorbent assay
Survey, replication 20 times draws the RLU values of 20 measurement results(Relative light unit), calculate its average value(M)And standard deviation
(SD), M+2SD is drawn, the luminous value is substituted into calibration curve calculating obtains corresponding concentration value.Using chemiluminescence detection side
The concentration value that method is obtained is 0.11 IU/mL, relative to traditional IU/mL of enzyme linked immunosorbent assay sensitivity for analysis 0.73, is carried
It is high about 7 times.
Claims (10)
1. a kind of glutamic acid decarboxylase antibody chemical luminescence immunity detection reagent, the kit includes:The glutamic acid of purifying
The coated magnetic particle working solution of decarboxylase, the glutamate decarboxylase working solution of the purifying of acridinium ester label, glutamate decarboxylase resist
Body calibration product, preexciting liquid, exciting liquid.
2. kit according to claim 1, it is characterised in that the coated solid phase carrier of glutamate decarboxylase is magnetic
Particulate.
3. kit according to claim 1, it is characterised in that the coated solid phase carrier of glutamate decarboxylase is carboxylic
The particle diameter of base is 0.05-1um magnetic particles.
4. kit according to claim 1, it is characterised in that the chemiluminescent labels are acridinium ester, acridinium ester
Sulfonamide, acridinium ester toluenesulfonamide, acridinium ester are to methylsulfonamides, acridinium ester trimethyl fluoride sulfonyl amine.
5. kit according to claim 1, it is characterised in that the preferred acridinium ester of chemiluminescent labels.
6. kit according to claim 1, it is characterised in that the Chemoluminescent substrate is excited including chemiluminescence
Liquid, chemiluminescence preexciting liquid.
7. kit according to claim 1, it is characterised in that the chemiluminescence preexciting liquid is mass fraction
0.005% ~ 0.5% hydrogen peroxide (H2O2) solution, exciting liquid is 0.005mol/L ~ 0.025mol/L sodium hydroxide
(NaOH) solution.
8. kit according to claim 1, it is characterised in that the glutamic acid decarboxylase antibody calibration product are to use standard
Savor buffer solution by glutamic acid decarboxylase antibody be configured to concentration for 5 IU/mL, 15 IU/mL, 35 IU/mL, 120 IU/mL,
250 IU/mL, 2000 IU/mL, packing are lyophilized, and 4 DEG C save backup.
9. kit according to claim 1, it is characterised in that the preparation method of the kit, it is characterised in that bag
Include the preparation, the preparation of the acridinium ester of glutamate decarboxylase mark, chemical luminous substrate of the coated magnetic particle of glutamate decarboxylase
The preparation of product is calibrated in the preparation of liquid, glutamic acid decarboxylase antibody.
10. the preparation method of kit according to claim 1 and claim 9, it is characterised in that comprise the following steps:
1)The preparation of the coated magnetic particle of glutamate decarboxylase:
The nanometer magnetic bead suspension of carboxylated is taken, Magneto separate removes supernatant, and MES buffer solutions are resuspended, and add the EDC aqueous solution, activate magnetic
Bead surface carboxyl, adds glutamate decarboxylase, at room temperature suspension 2-10 h, Magneto separate, removes supernatant, and Tris buffer solutions are resuspended,
Obtain the coated magnetic particle of glutamate decarboxylase;Optionally, a diameter of 0.1 μm ~ 2.0 μm of carboxylated nanometer magnetic bead;MES is buffered
Liquid concentration is 10mM ~ 100mM, pH 5.5 ~ 8.5;
2)The preparation of the acridinium ester of glutamate decarboxylase mark:
Glutamate decarboxylase is taken, carbonate buffer solution is added, mixed, acridinium ester is then added and mixes, lucifuge is reacted at room temperature, 1-
Taken out after 2 h, centrifuge and handled respectively with pure water and TBS buffer solutions first in desalting column desalting processing, desalination processes, most
The acridine ester solution of obtained glutamate decarboxylase mark is added afterwards, and liquid to the preservation collected in centrifuge tube is in control glutamic acid
The acridinium ester of decarboxylase mark;
3)Glutamic acid decarboxylase antibody calibrates the preparation of product:
With standard items buffer solution by glutamic acid decarboxylase antibody be configured to concentration for 5 IU/mL, 20 IU/mL, 100 IU/mL,
400 IU/mL, 800 IU/mL, 2000 IU/mL, packing are lyophilized, and 4 DEG C save backup;
4)The preparation of chemiluminescence preexciting liquid:
1.0 liters of purified waters are measured, the hydrogen peroxide (H that 0.5 ~ 100uL mass fractions are 20% is sequentially added2O2), 0.5 ~ 5 gram prevent
Rotten agent, 0.5 ~ 5 gram of surfactant, shake up rear lucifuge storage;Optionally, preservative be commercialization sodium azide, PC300,
Surfactant is polysorbas20, Tween 80, Triton X-100, Triton 405;
5)The preparation of chemiluminescence exciting liquid:
1.0 liters of purified waters are measured, 0.2 ~ 1 gram of sodium hydroxide, 0.5 ~ 5 gram of preservative, 0.5 ~ 5 gram of surface is sequentially added and lives
Property agent, shake up the storage of rear lucifuge;Optionally, preservative is commercialization sodium azide, PC300, and surfactant is polysorbas20, told
Temperature 80, Triton X-100, TritonX- 405.
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108414766A (en) * | 2018-01-29 | 2018-08-17 | 上海良润生物医药科技有限公司 | Kit for quantitatively detecting diabetes autoantibody and its application |
WO2020082720A1 (en) * | 2018-10-25 | 2020-04-30 | 成都博奥新景医学科技有限公司 | Method and system for single-part chemiluminescence immunoassay on basis of magnetic rod method |
CN114924079A (en) * | 2022-03-29 | 2022-08-19 | 北京世纪沃德生物科技有限公司 | Anti-glutamate decarboxylase antibody determination kit and detection method |
-
2016
- 2016-11-21 CN CN201611018770.4A patent/CN107102140A/en active Pending
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108414766A (en) * | 2018-01-29 | 2018-08-17 | 上海良润生物医药科技有限公司 | Kit for quantitatively detecting diabetes autoantibody and its application |
WO2020082720A1 (en) * | 2018-10-25 | 2020-04-30 | 成都博奥新景医学科技有限公司 | Method and system for single-part chemiluminescence immunoassay on basis of magnetic rod method |
CN114924079A (en) * | 2022-03-29 | 2022-08-19 | 北京世纪沃德生物科技有限公司 | Anti-glutamate decarboxylase antibody determination kit and detection method |
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