CN103116030A - Kit and method for detecting autoimmune antibody of type-I diabetes mellitus - Google Patents
Kit and method for detecting autoimmune antibody of type-I diabetes mellitus Download PDFInfo
- Publication number
- CN103116030A CN103116030A CN201310035563XA CN201310035563A CN103116030A CN 103116030 A CN103116030 A CN 103116030A CN 201310035563X A CN201310035563X A CN 201310035563XA CN 201310035563 A CN201310035563 A CN 201310035563A CN 103116030 A CN103116030 A CN 103116030A
- Authority
- CN
- China
- Prior art keywords
- reagent
- antibody
- luciferase
- fusion
- kit
- 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
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a kit and method for detecting an autoimmune antibody of the type-I diabetes mellitus and belongs to the technical field of biochemical medicine detection. The method comprises the following steps of: adding a luciferase as an antibody fusion protein, wherein the luciferase as the antibody fusion protein is generated by 293 cell culture and can be specifically combined with a diabetes mellitus autoantibody in serum of a patient; then, adding protein-A agarose, depositing a fusion protein as an antibody compound, centrifuging, and then, absorbing the uncombined fusion protein from a supernatant liquid; and then, adding a luciferase substrate, and detecting the fluorescence intensity by using a fluorescence detection instrument to finally measure the content of the diabetes mellitus autoantibody in a sample to be detected. Compared with the traditional HPLC (High Performance Liquid Chromatography), a micro-quantitative fluorescence detection instrument used for detecting in the method has the advantages of simplicity in operation, no need of uncovering to block a pollution way, high sensitivity and signal to noise ratio, stable and reliable measured value and capability of ensuring reliable experiment result and safety of operating personnel and meeting the requirements of micro quantity and regent saving.
Description
Technical field
The present invention relates to biochemical medicine inspection technology field, specifically a kind of detection type i diabetes autoimmune antibody kit and detection method thereof, can be used for prediction and the diagnosis of insulin-dependent diabetes mellitus (IDDM).
Background technology
General, type i diabetes is a kind of autoimmune disease, body is infected the factors such as (especially virus infections), poisonous substance and brings out and produce abnormal self body fluid and cellullar immunologic response, cause the beta Cell of islet damage, insulin secretion reduces, in case needing to rely on exogenous insulin, morbidity replenishes to sustain life, so insulin-dependent diabetes mellitus (IDDM) claims again insulin-dependent diabetes.
This paradiabetes is common in children and young patient, but it can occur in the crowd of any age bracket, and the morbidity rate of this paradiabetes accounts for 10% of total diabetes cases.As seen insulin-dependent diabetes mellitus (IDDM) is diagnosed early and the people at highest risk in forecast and have great importance.
The diagnosis of type i diabetes is mainly passed through to detect himself immune antibody, and antibody mainly comprises following several: glutamic acid decarboxylase antibody (GAD), insular cellular antibody (ICA) tyrosine phosphatase enzyme antibody (IA-2A) and insulin autoimmune antibody (IAA).Present mensuration GAD, IA-2, the method that IAA is commonly used have radioimmunology, euzymelinked immunosorbent assay (ELISA); Be used for the main indirect immunofluorescence of using that ICA measures, in addition also useful euzymelinked immunosorbent assay (ELISA) and Immunohistochemical Method.
Radiommunoassay has the unrivaled advantage of many other analytical approachs.It had both had immunoreactive high specific, had again radiometric high sensitivity, and amount of samples is few, often can survey to the picomole amount.This method occur sometimes cross reaction, false positive reaction, tissue sample process rapid not, can not deactivation digestive enzyme and salt, affect sometimes result etc., also have problems such as using the caused radiation of radioactively labelled substance and pollution.
Summary of the invention
Technical assignment of the present invention is to solve the deficiencies in the prior art, and a kind of detection type i diabetes autoimmune antibody kit and detection method thereof are provided.
Technical scheme of the present invention realizes in the following manner, and the present invention utilizes 293 cells to cultivate a kind of luciferase-antigen coalescence protein that produces, can with the patients serum in diabetes autoantibody generation specific binding.Then add the albumin A agarose, precipitation fusion-antibody complex siphons away unconjugated fusion after centrifugal from supernatant.Add afterwards the luciferase substrate, utilize the fluorescence detector fluorescence intensity, thereby measure the content of diabetes autoantibody in testing sample.
A kind of detection type i diabetes autoimmune antibody kit, this kit includes: the fusion of luciferase and diabetes antigen.
A kind of detection type i diabetes autoimmune antibody kit includes: luciferase-GAD65 fusion, luciferase-IA-2 fusion, luciferase-IA-2 beta fusion proteins.
A kind of detection type i diabetes autoimmune antibody kit includes:
Reagent 1:
1A: luciferase-GAD65 fusion,
1B: luciferase-IA-2 fusion,
1C: luciferase-IA-2 beta fusion proteins;
Reagent 2: titer;
Reagent 3:
3A: blank liquid,
3B: Healthy Human Serum contrast liquid;
Reagent 4: albumin A agarose;
Reagent 5:Ranilla fluorogenic substrate.
The preparation method of described reagent 1 is: will be transfected in 293 cells after Ranilla fluorogene and genes of interest fusion, then cultivate 36～60h in containing the DMEM nutrient solution of 10% serum, cell ultrasonic processing 15～30 min in ice bath with results, then with cell pyrolysis liquid in 4 ℃, to get supernatant after centrifugal 15～45 min of 6000～8000rpm rotating speed, obtain fusion, rear packing is lyophilized into the reagent 1 in this kit.
Titer is standard antibody NIBSC97/550.
Damping fluid refers to 0.01mol/L, the pH7.4 phosphate buffer.
The Ranilla luciferase refers to: renilla luciferase refers to that the reporter gene of luciferase comes from sea pansy.Just as firefly luciferase.
The Ranilla fluorogene comes from sea pansy.
Genes of interest refers to the diabetes antigen gene.With expressing after these two kinds of gene fusion, can produce fusion, be respectively luciferase-GAD-65 fusion, luciferase-IA-2 fusion, luciferase-IA-2 beta fusion proteins in this patent.
A kind of step that detects the detection method of type i diabetes autoimmune antibody kit is:
Kit:
Reagent 1:
1A: luciferase-GAD65 fusion,
1B: luciferase-IA-2 fusion,
1C: luciferase-IA-2 beta fusion proteins;
Reagent 2: titer;
Reagent 3:
3A: blank liquid,
3B: Healthy Human Serum contrast liquid;
Reagent 4: albumin A agarose;
Reagent 5:Ranilla fluorogenic substrate;
Before experiment, kit and damping fluid are at room temperature placed 30min at least;
(1) reagent 1 use damping fluid is carried out the gradient dilution of certain multiple, guarantee that antigen has certain concentration, requiring to reach in every microlitre has 10
7Individual flat fluorescent;
(2) reagent 2 titers, Healthy Human Serum contrast liquid that add patients serum, the gradient dilution of 15～30uL and arrange N test tube as parallel group (concentration gradient of mark song can according to the different set of antibody) in N centrifuge tube;
(3) to every reagent 1 dilution that adds 40～80uL in by all means, add in addition two blank pipes and be used for surveying the fluorescence total intensity;
(4) with the every arm of vortex oscillation device mixing and close the lid, do not comprise that two are surveyed fluorescence total intensity blank pipes;
(5) at room temperature, hatch 3～5h, allow diabetes autoantibody be combined with antigentic specificity;
(6) hatch after, to every albumin A agarose that adds 40～80 uL in by all means, do not comprise and survey the total fluorescence intensity blank pipe, come the capture antigen antibody complex;
(7) with the every arm of vortex oscillation device mixing and close the lid, do not comprise and survey the total fluorescence intensity blank pipe;
(8) slowly shake the antigen-antibody mixture overnight or slowly shake antigen-antibody potpourri 1～2h in room temperature in 4 ℃, make antibody and albumin A agarose coupled,
(9) hatch after, to every damping fluid that adds 2～8 ℃ of refrigerations of 1～2 mL in by all means, do not comprise and survey the total fluorescence intensity blank pipe, the centrifugal 15～30min of 1500～3000rpm under 4 ℃ then, the antigen-antibody complex that the albumin A agarose is coupled is centrifugal to managing the end;
(10) after centrifugal, supernatant is carefully sucked, precipitation has been hanged centrifuge washing 2～5 times with PBS with albumin A agarose-antigen-antibody complex;
(11) then add 50 uL Ranilla fluorogenic substrates in centrifuge tube, measure immediately fluorescence intensity;
(12) analysis result:
Take the concentration of titer as X-axis, to do typical curve in conjunction with rate as Y-axis, then according to patients serum's fluorescence intensity, the calculations incorporated rate reads the concentration of autoantibody the patients serum from curve.
In conjunction with rate=(sample determination value-3B measured value)/total fluorescence intensity
The relative multiple of the bent read value * of antibody concentration=mark
Negative :≤1.0 U/mL
Positive: ﹥ 1.0 U/mL
As long as three kinds of antibody have a kind of test positive, can determine that sample contains diabetes autoantibody, the sample source human body suffers from I type diabetes.
The beneficial effect that the present invention compared with prior art produces is:
The present invention uses the fluorescein fusion thing that serves as a mark on the basis of radioimmunoassay technique, make kit, and the application of sample program is simple, once can analyze a large amount of samples, and has higher specificity and sensitivity.
The present invention uses micro-fluorescence detector to detect, and compares with traditional HPLC, and simple to operate, directly Acceptable criterion 0.2mL centrifuge tube is isolated mensuration, need not to uncap to intercept pollution channel, guarantees the reliable and operator safety of experimental result.Can adopt in addition to be not less than the 25uL sample size and to measure, reach the requirement that trace is economized reagent, and high sensitivity, high s/n ratio, measured value is reliable and stable.
Embodiment
The below is described in detail below a kind of detection type i diabetes autoimmune antibody kit of the present invention and detection method thereof.
A kind of luciferase-antigen coalescence protein that the present invention cultivate to produce with 293 cells, can with the patients serum in diabetes autoantibody generation specific binding.Then add the albumin A agarose, precipitation fusion-antibody complex siphons away unconjugated fusion after centrifugal from supernatant.Add afterwards the luciferase substrate, utilize the fluorescence detector fluorescence intensity, thereby measure the content of diabetes autoantibody in testing sample.
A kind of step that detects the detection method of type i diabetes autoimmune antibody kit of the present invention is:
(1) Ranilla fluorogene and genes of interest are transfected in 293 cells, then cultivate 55h in containing the DMEM nutrient solution of 10% serum; With results cell ultrasonic processing 1 min in ice bath, then with cell pyrolysis liquid in 4 ℃, to get supernatant after centrifugal 20 min of 6000～8000rpm rotating speed, rear packing is lyophilized into detection kit reagent 1;
Kit:
Reagent 1:
1A: luciferase-GAD65 fusion,
1B: luciferase-IA-2 fusion,
1C: luciferase-IA-2 beta fusion proteins;
Reagent 2: titer;
Reagent 3:
3A: blank liquid,
3B: Healthy Human Serum contrast liquid;
Reagent 4: albumin A agarose;
Reagent 5:Ranilla fluorogenic substrate;
Titer is standard antibody NIBSC97/550.
Damping fluid refers to 0.01mol/L, the pH7.4 phosphate buffer.
The Ranilla luciferase refers to: renilla luciferase refers to that the reporter gene of luciferase comes from sea pansy.Just as firefly luciferase.
The Ranilla fluorogene comes from sea pansy.
Genes of interest refers to the diabetes antigen gene.With expressing after these two kinds of gene fusion, can produce fusion, be respectively luciferase-GAD-65 fusion, luciferase-IA-2 fusion, luciferase-IA-2 beta fusion proteins in this patent.
The reagent of kit forms and pre-service:
Reagent numbering: 1A
Reagent name: luciferase-GAD65 fusion
Reagent numbering: 1B
Reagent name: luciferase-IA-2 fusion
Reagent numbering: 1C
Reagent name: luciferase-IA-2 beta fusion proteins
Pretreatment mode: freeze-dried powder: add the damping fluid of certain volume to dissolve mixing in every bottle, 2-8 ℃ of preservation is stand-by.
Reagent numbering: 2
Reagent name: titer
Pretreatment mode: utilize progressively gradient dilution titer, take fluorescence intensity as ordinate, take the concentration of standard antibody NIBSC97/550 as horizontal ordinate, do typical curve.
Reagent numbering: 3A
Reagent name: blank liquid
Reagent numbering: 3B
Reagent name: Healthy Human Serum contrast liquid
Pretreatment mode: 2-8 ℃ of preservation is stand-by.
Reagent numbering: 4
Reagent name: albumin A agarose
Pretreatment mode: freeze-dried powder, add the damping fluid of certain volume to dissolve mixing in every bottle, 2-8 ℃ of preservation is stand-by.
Reagent numbering: 5
Reagent name: Ranilla fluorogenic substrate
Pretreatment mode: 2-8 ℃ of preservation is stand-by.
(2) titer, contrast liquid that add patients serum, the gradient dilution of 20uL and arrange N test tube as parallel group in N centrifuge tube;
(3) to every fusion dilution that adds 50uL in by all means, add in addition two blank pipes and be used for surveying the fluorescence total intensity;
(4) with the every arm of vortex oscillation device mixing and close the lid, do not comprise and survey the total fluorescence intensity blank pipe;
(5) at room temperature, hatch 5h, allow diabetes autoantibody be combined with antigentic specificity;
(6) hatch after, to every albumin A agarose that adds 50 uL in by all means, do not comprise and survey the total fluorescence intensity blank pipe, come the capture antigen antibody complex;
(7) with the every arm of vortex oscillation device mixing and close the lid, do not comprise and survey the total fluorescence intensity blank pipe;
(8) 4 ℃ are slowly shaken antigen-antibody mixture overnight or room temperature 1h, and antibody is combined with the albumin A agarose;
(9) hatch after, to every damping fluid that adds 2～8 ℃ of refrigerations of 1 mL in by all means, do not comprise and survey the total fluorescence intensity pipe, then at 4 ℃ of centrifugal 30min of lower 1500rpm, the antigen-antibody complex that the albumin A agarose is coupled is centrifugal to managing the end;
(10) after centrifugal, supernatant is carefully sucked, precipitation has been hanged centrifuge washing 2～5 times with PBS with albumin A agarose-antigen-antibody complex;
(11) then add 50 uL Ranilla fluorogenic substrates in centrifuge tube, measure immediately fluorescence intensity;
(12) analysis result:
Take the concentration of titer as X-axis, to do typical curve in conjunction with rate as Y-axis, then according to patients serum's fluorescence intensity, the calculations incorporated rate reads the concentration of autoantibody the patients serum from curve.
In conjunction with rate=(sample determination value-3B measured value)/total fluorescence intensity
The relative multiple of the bent read value * of antibody concentration=mark
Negative :≤1.0 U/mL
Positive: ﹥ 1.0 U/mL
As long as three kinds of antibody have a kind of test positive, can determine that sample contains diabetes autoantibody, the sample source human body suffers from type i diabetes.
Respectively 107 Healthy Peoples and 98 serum that have been diagnosed as the patient of type i diabetes are detected by above-described embodiment, its result is as follows:
The testing result of 1:107 position Healthy Human Serum
Antibody: GAD-65 negative rate: 100%
Antibody: IA-2 negative rate: 99%
Antibody: IA-2 β negative rate: 99%.
2:98 position type i diabetes patients serum's testing result
Antibody: GAD-65 positive rate: 75%
Antibody: IA-2 positive rate: 53%
Antibody: IA-2 β positive rate: 56%.
3: utilize 2005 double antibody solid phase method research the method, specificity and sensitivity result are as follows:
The specificity of the method and sensitivity result:
Antibody: GAD-65 specificity (n=100): 96% sensitivity (n=50): 87%
Antibody: IA-2 specificity (n=100): 100% sensitivity (n=50): 73%
Antibody: IA-2 β specificity (n=100): 98% sensitivity (n=50): 74%.
Claims (6)
1. one kind is detected type i diabetes autoimmune antibody kit, it is characterized in that this kit includes: the fusion of luciferase and diabetes antigen.
2. one kind is detected type i diabetes autoimmune antibody kit, it is characterized in that this kit includes: luciferase-GAD65 fusion, luciferase-IA-2 fusion, luciferase-IA-2 beta fusion proteins.
3. one kind is detected type i diabetes autoimmune antibody kit, it is characterized in that this kit includes:
Reagent 1:
1A: luciferase-GAD65 fusion,
1B: luciferase-IA-2 fusion,
1C: luciferase-IA-2 beta fusion proteins;
Reagent 2: titer;
Reagent 3:
3A: blank liquid,
3B: Healthy Human Serum contrast liquid;
Reagent 4: albumin A agarose;
Reagent 5:Ranilla fluorogenic substrate.
4. a kind of detection type i diabetes autoimmune antibody kit according to claim 3, the preparation method who it is characterized in that described reagent 1 is: will be transfected in 293 cells after Ranilla fluorogene and genes of interest fusion, then cultivate 36～60h in containing the DMEM nutrient solution of 10% serum, cell ultrasonic processing 15～30 min in ice bath with results, then with cell pyrolysis liquid in 4 ℃, to get supernatant after centrifugal 15～45 min of 6000～8000rpm rotating speed, obtain fusion, rear packing is lyophilized into the reagent 1 in this kit.
5. detection method that detects type i diabetes autoimmune antibody kit is characterized in that the step of this detection method is:
Kit:
Reagent 1:
1A: luciferase-GAD65 fusion,
1B: luciferase-IA-2 fusion,
1C: luciferase-IA-2 beta fusion proteins;
Reagent 2: titer;
Reagent 3:
3A: blank liquid,
3B: Healthy Human Serum contrast liquid;
Reagent 4: albumin A agarose;
Reagent 5:Ranilla fluorogenic substrate;
Before experiment, kit and damping fluid are at room temperature placed 30min at least;
(1) reagent 1 use damping fluid is carried out the gradient dilution of certain multiple, guarantee that antigen has certain concentration, requiring to reach in every microlitre has 10
7Individual flat fluorescent;
(2) reagent 2 titers, Healthy Human Serum contrast liquid that add patients serum, the gradient dilution of 15～30uL and arrange N test tube as parallel group (concentration gradient of mark song can according to the different set of antibody) in N centrifuge tube;
(3) to every reagent 1 dilution that adds 40～80uL in by all means, add in addition two blank pipes and be used for surveying the fluorescence total intensity;
(4) with the every arm of vortex oscillation device mixing and close the lid, do not comprise that two are surveyed fluorescence total intensity blank pipes;
(5) at room temperature, hatch 3～5h, allow diabetes autoantibody be combined with antigentic specificity;
(6) hatch after, to every albumin A agarose that adds 40～80 uL in by all means, do not comprise and survey the total fluorescence intensity blank pipe, come the capture antigen antibody complex;
(7) with the every arm of vortex oscillation device mixing and close the lid, do not comprise and survey the total fluorescence intensity blank pipe;
(8) slowly shake the antigen-antibody mixture overnight or slowly shake antigen-antibody potpourri 1～2h in room temperature in 4 ℃, make antibody and albumin A agarose coupled,
(9) hatch after, to every damping fluid that adds 2～8 ℃ of refrigerations of 1～2 mL in by all means, do not comprise and survey the total fluorescence intensity blank pipe, the centrifugal 15～30min of 1500～3000rpm under 4 ℃ then, the antigen-antibody complex that the albumin A agarose is coupled is centrifugal to managing the end;
(10) after centrifugal, supernatant is carefully sucked, precipitation has been hanged centrifuge washing 2～5 times with PBS with albumin A agarose-antigen-antibody complex;
(11) then add 50 uL Ranilla fluorogenic substrates in centrifuge tube, measure immediately fluorescence intensity;
(12) analysis result.
6. detection method that detects type i diabetes autoimmune antibody kit is characterized in that the step of this detection method is:
(1) Ranilla fluorogene and genes of interest are transfected in 293 cells, then cultivate 55h in containing the DMEM nutrient solution of 10% serum; With results cell ultrasonic processing 1 min in ice bath, then with cell pyrolysis liquid in 4 ℃, to get supernatant after centrifugal 20 min of 6000～8000rpm rotating speed, rear packing is lyophilized into detection kit reagent 1;
Kit:
Reagent 1:
1A: luciferase-GAD65 fusion,
1B: luciferase-IA-2 fusion,
1C: luciferase-IA-2 beta fusion proteins;
Reagent 2: titer;
Reagent 3:
3A: blank liquid,
3B: Healthy Human Serum contrast liquid;
Reagent 4: albumin A agarose;
Reagent 5:Ranilla fluorogenic substrate;
(2) titer, contrast liquid that add patients serum, the gradient dilution of 20uL and arrange N test tube as parallel group in N centrifuge tube;
(3) to every fusion dilution that adds 50uL in by all means, add in addition two blank pipes and be used for surveying the fluorescence total intensity;
(4) with the every arm of vortex oscillation device mixing and close the lid, do not comprise and survey the total fluorescence intensity blank pipe;
(5) at room temperature, hatch 5h, allow diabetes autoantibody be combined with antigentic specificity;
(6) hatch after, to every albumin A agarose that adds 50 uL in by all means, do not comprise and survey the total fluorescence intensity blank pipe, come the capture antigen antibody complex;
(7) with the every arm of vortex oscillation device mixing and close the lid, do not comprise and survey the total fluorescence intensity blank pipe;
(8) 4 ℃ are slowly shaken antigen-antibody mixture overnight or room temperature 1h, and antibody is combined with the albumin A agarose;
(9) hatch after, to every damping fluid that adds 2～8 ℃ of refrigerations of 1 mL in by all means, do not comprise and survey the total fluorescence intensity pipe, then at 4 ℃ of centrifugal 30min of lower 1500rpm, the antigen-antibody complex that the albumin A agarose is coupled is centrifugal to managing the end;
(10) after centrifugal, supernatant is carefully sucked, precipitation has been hanged centrifuge washing 2～5 times with PBS with albumin A agarose-antigen-antibody complex;
(11) then add 50 uL Ranilla fluorogenic substrates in centrifuge tube, measure immediately fluorescence intensity;
(12) analysis result.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310035563XA CN103116030A (en) | 2013-01-30 | 2013-01-30 | Kit and method for detecting autoimmune antibody of type-I diabetes mellitus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310035563XA CN103116030A (en) | 2013-01-30 | 2013-01-30 | Kit and method for detecting autoimmune antibody of type-I diabetes mellitus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103116030A true CN103116030A (en) | 2013-05-22 |
Family
ID=48414457
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310035563XA Pending CN103116030A (en) | 2013-01-30 | 2013-01-30 | Kit and method for detecting autoimmune antibody of type-I diabetes mellitus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103116030A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103969234A (en) * | 2014-04-17 | 2014-08-06 | 山东东兴汇智生物科技有限公司 | Method capable of detecting four kinds of type I diabetes autoimmune antibodies simultaneously |
| CN105579853A (en) * | 2013-10-17 | 2016-05-11 | 奥古斯特·皮·松耶尔生物医学研究所 | Diagnostic method for detecting a GABA(A) related autoimmune disease and related subject-matter |
| CN107884371A (en) * | 2017-04-28 | 2018-04-06 | 南方医科大学 | Luciferase immuno absorbence method for high flux antibody quick detection |
| CN108318689A (en) * | 2018-04-09 | 2018-07-24 | 北京大学深圳研究生院 | A kind of diagnostic method of Huppert's disease |
| CN108593912A (en) * | 2018-04-09 | 2018-09-28 | 北京大学深圳研究生院 | A kind of detection method of solubility CD38 concentration |
| CN109752547A (en) * | 2018-12-17 | 2019-05-14 | 杭州京北生物科技有限公司 | A kind of breast cancer autoimmune antibody detection kit and the preparation method and application thereof |
| CN111175518A (en) * | 2020-01-08 | 2020-05-19 | 杭州北角医学检验所有限公司 | Combined antibody detection kit for type I diabetes and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101057976A (en) * | 2002-08-06 | 2007-10-24 | 洛马林达大学 | Substances for preventing and treating autoimmune diseases |
| CN101277722A (en) * | 2005-08-06 | 2008-10-01 | 王庆华 | Composition and method for prevention and treatment of type I diabetes |
| CN101825637A (en) * | 2009-03-03 | 2010-09-08 | 中国人民解放军军事医学科学院基础医学研究所 | Composition for immunologically diagnosing type I diabetes |
-
2013
- 2013-01-30 CN CN201310035563XA patent/CN103116030A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101057976A (en) * | 2002-08-06 | 2007-10-24 | 洛马林达大学 | Substances for preventing and treating autoimmune diseases |
| CN101277722A (en) * | 2005-08-06 | 2008-10-01 | 王庆华 | Composition and method for prevention and treatment of type I diabetes |
| CN101825637A (en) * | 2009-03-03 | 2010-09-08 | 中国人民解放军军事医学科学院基础医学研究所 | Composition for immunologically diagnosing type I diabetes |
Non-Patent Citations (2)
| Title |
|---|
| PETER D.BURBELO, ET AL.: "A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2", 《DIABETES CARE》, vol. 31, no. 9, 5 June 2008 (2008-06-05), pages 1824 - 1826 * |
| PETER D.BURBELO, ET AL.: "Comparison of Radioimmunoprecipitation With Luciferase Immunoprecipitation for Autoantibodies to GAD65 and IA-2β", 《DIABETES CARE》, vol. 33, no. 4, 19 January 2010 (2010-01-19), pages 754 - 756 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105579853A (en) * | 2013-10-17 | 2016-05-11 | 奥古斯特·皮·松耶尔生物医学研究所 | Diagnostic method for detecting a GABA(A) related autoimmune disease and related subject-matter |
| CN103969234A (en) * | 2014-04-17 | 2014-08-06 | 山东东兴汇智生物科技有限公司 | Method capable of detecting four kinds of type I diabetes autoimmune antibodies simultaneously |
| CN103969234B (en) * | 2014-04-17 | 2017-02-15 | 山东东兴汇智生物科技有限公司 | Luciferase- poly-antigen fusion protein and protein A agarose-fusion protein-antibody complex |
| CN107884371A (en) * | 2017-04-28 | 2018-04-06 | 南方医科大学 | Luciferase immuno absorbence method for high flux antibody quick detection |
| CN108318689A (en) * | 2018-04-09 | 2018-07-24 | 北京大学深圳研究生院 | A kind of diagnostic method of Huppert's disease |
| CN108593912A (en) * | 2018-04-09 | 2018-09-28 | 北京大学深圳研究生院 | A kind of detection method of solubility CD38 concentration |
| CN109752547A (en) * | 2018-12-17 | 2019-05-14 | 杭州京北生物科技有限公司 | A kind of breast cancer autoimmune antibody detection kit and the preparation method and application thereof |
| CN111175518A (en) * | 2020-01-08 | 2020-05-19 | 杭州北角医学检验所有限公司 | Combined antibody detection kit for type I diabetes and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103116030A (en) | Kit and method for detecting autoimmune antibody of type-I diabetes mellitus | |
| Hage | Immunoassays | |
| JP2505979B2 (en) | Immunoassay device | |
| CN102023211A (en) | Immunochromatographic test strip for full quantitative detection of C-reactive protein and preparation method thereof | |
| CN104198725A (en) | Anti-cyclic citrullinated peptide (CCP) antibody detection kit | |
| CN108152512A (en) | Heparin-binding protein detection kit and preparation method thereof | |
| CN104655846A (en) | Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof | |
| CN102841206B (en) | TnT (Troponin-T, TNT) measures kit | |
| CN103454431A (en) | Immunoturbidimetric kit for detecting beta2-microglobulin and preparation method thereof | |
| CN103063851A (en) | Free triiodothyronine nanometer magnetic particle chemiluminescence assay kit and preparation method thereof and detection method thereof | |
| CN108318690A (en) | A kind of immunofluorescence chromatographic test paper and its preparation method and application | |
| CN103969234B (en) | Luciferase- poly-antigen fusion protein and protein A agarose-fusion protein-antibody complex | |
| CN105785041A (en) | Test strip for quantitatively detecting calprotectin, preparation method thereof and determining method for calprotectin concentration | |
| CN106324251B (en) | The preparation method and beta 2-microglobulin detecting kit of small fragment BMG antibody | |
| EP3243076A1 (en) | Methods for detecting a marker for active tuberculosis | |
| CN204989196U (en) | Qualitative test paper strip of immunity chromatography is united to six antinuclear antibodiess | |
| CN104897864A (en) | ELISA (enzyme linked immunosorbent assay) kit for detecting amantadine and application of ELISA kit | |
| CN103048471A (en) | Method for quantitatively detecting protein acetylation level | |
| CN105785012A (en) | Enzyme linked immunosorbent assay kit for detecting ribavirin and application thereof | |
| CN103645321A (en) | Test paper for screening procalcitonin and preparation method of test paper | |
| CN109298178A (en) | Cardiac myosin binding protein C(cMyBP-C based on immunomagnetic beads) time-resolved fluoroimmunoassay kit | |
| CN103048477A (en) | Nanometer magnetic particle chemiluminescence detection kit for triiodothyronine as well as preparation method and detecting method of same | |
| CN106546729B (en) | Novel process method for removing serum matrix effect in dry immunofluorescence quantitative detection | |
| CN113030456A (en) | Detection test strip, detection card and kit for anti-cyclic citrullinated peptide antibody | |
| CN206038688U (en) | Immunity chromatography detection test strip's fluorescent quantitation spectral detection system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20130522 |