CN105445257A - Kit and method for detecting allergen-specific IgE antibody of cat hair - Google Patents
Kit and method for detecting allergen-specific IgE antibody of cat hair Download PDFInfo
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- CN105445257A CN105445257A CN201510788502.XA CN201510788502A CN105445257A CN 105445257 A CN105445257 A CN 105445257A CN 201510788502 A CN201510788502 A CN 201510788502A CN 105445257 A CN105445257 A CN 105445257A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
Abstract
The invention discloses a kit and a method for detecting the allergen-specific IgE antibody of cat hair. The kit comprises a calibration product, a quality control product, a biotin-labeled cat hair allergen solution, an alkaline phosphatase labeled mouse anti-human IgE secondary antibody solution and streptavidin labeled nano magnetic micro particle suspension; the kit is used for detecting the allergen-specific IgE antibody of cat hair. By the mode above, each reagent component in the kit is good in stability, and the period of validity can be more than or equal to one year; the detection flexibility is high, the specificity is good, and variation is tiny. A perfect uniform technology is obtained via the technological optimization of a great amount of experiments, and production is carried out according to the standardized production operation instructions and quality control instructions vigorously. A user can perform standard operation according to the operation specification so as to obtain the reliable result. In the clinical study, the conforming relevance to imported reagents abroad is higher than or equal to 90%, and the expense is only 1/2 that of the imported reagents abroad.
Description
Technical field
The invention belongs to the outer diagnostic field of medicine equipment biological species immune body, particularly relate to a kind of kit and method of cat hair allergenic specific IgE antibody.
Background technology
The type i allergic reaction disease of IgE antibody mediation is quite general, the total incidence of countries in the world allergic disease is about 15%-30%, the people of the nearly 5-10% of China is subject to the harassment of anaphylactogen, the great hygienic problems of our times, by one of the World Health Organization's three large diseases being classified as 21st century keypoint control.
Anaphylactia is that patient sucks, ingesting into or injecting the material containing allergen (is called anaphylactogen or allergen, Allergen) B cell triggering body afterwards produces Serum specificity immunoglobulin E (ImmunoglobulinE, IgE), thus initiation allergic reaction (or claim allergic reaction, Allergy) disease and related symptoms, as allergic asthma, hay fever, nettle rash, allergic rhinitis, eczema, conjunctivitis and intestines and stomach I type anaphylactia and severe allergic reaction etc.The generation of above-mentioned anaphylactia, IgE antibody plays a crucial role.
At home, cat hair allergenic specific IgE antibody (E1) detection is main is clinically applied as master with external import reagent and SABC reagent, and external import reagent price is very expensive, brings very large financial burden to patient, is unfavorable for popularizing in basic unit.Domestic cat hair allergenic specific IgE antibody (E1) the immunochemiluminescence detection kit with independent intellectual property right does not also have at present.
Therefore, the detection technique that also can reduce testing cost to cat hair allergenic specific IgE antibody (E1) detection sensitivity height and reliability to be developed is had.
Summary of the invention
The technical matters that the present invention mainly solves is to provide a kind of kit and the method that detect cat hair allergenic specific IgE antibody, this kit dependable performance, highly sensitive, the range of linearity is wide, full-automatic instrument can be coordinated to use, detection sensitivity and reliability can be improved, and reduce costs, extend the expiration date.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: provide a kind of kit detecting cat hair allergenic specific IgE antibody, the reagent comprised has: calibration object, quality-control product, biotin labeled cat hair allergen solution, the anti-solution of mouse anti human IgE two of alkali phosphatase enzyme mark, the nano magnetic microparticle suspending liquid of marked by streptavidin.
In a preferred embodiment of the present invention, the concentration of described biotin labeled cat hair allergen solution is 0.1 ~ 1.0ug/ml, the concentration of the nano magnetic microparticle suspending liquid of described marked by streptavidin is 0.1 ~ 1.0mg/ml, and the concentration of the anti-solution of mouse anti human IgE two of described alkali phosphatase enzyme mark is 0.1 ~ 2.0ug/ml.
In a preferred embodiment of the present invention, the concentration of described calibration object is: 0IU/ml, 0.35IU/ml, 0.7IU/ml, 3.5IU/ml, 17.5IU/ml, 50IU/ml, 100IU/ml, described calibration object to be IgE albumen and 0.1M, pH value be 7.4 Tris-HCl buffer obtain; The concentration of described quality-control product is: 0.7IU/ml, 17.5IU/ml, described quality-control product to be IgE albumen and 0.1M, pH value be 7.4 Tris-HCl buffer obtain.
In a preferred embodiment of the present invention, described cat hair anaphylactogen be can with the protein bound antigen of cat hair specific IgE in human body, the preparation process of described biotin labeled cat hair anaphylactogen is: joined by 1:20 mol ratio by cat hair anaphylactogen and be dissolved in the biotin of dimethyl formamide, then obtains with the phosphate buffer dialysis that 0.1M, pH value are 7.4.
In a preferred embodiment of the present invention, the mouse anti human IgE two of described mark resist for can with the antibody of IgE albumen specific bond in human body, the preparation process that the mouse anti human IgE two of described alkali phosphatase enzyme mark is anti-is: the alkaline phosphatase enzyme solutions by concentration being 1.0-5.0mg/mL is anti-in molar ratio for 2-10:1 carries out mixing rear also purifying with mouse anti human IgE two, the mouse anti human IgE two obtaining alkali phosphatase enzyme mark resists, described purifying balances and wash-out with the bicarbonate buffer that pH is 8-9, UV detect and record purifying collection of illustrative plates again, use 0.05M again, pH value be 6.0 MES damping fluid the mouse antihuman IgE antibody of alkali phosphatase enzyme mark is diluted.
In a preferred embodiment of the present invention, the preparation process of the nano magnetic microparticle suspending liquid of described marked by streptavidin is: precipitated by the nano magnetic particulate magnetic separator of marked by streptavidin, again with 0.01M, pH value be 7.4 phosphate buffer resuspended, mixing Magneto separate precipitation, and repeated washing, the nano magnetic microparticulate after cleaning in 0.01M, pH value be the phosphate buffer of 7.4.
There is provided a kind of nanometer Magnetism particulate immuno chemistry luminescence method quantitatively to detect the method for serum cat hair allergenic specific IgE antibody, comprising step is:
(1) magnetic particle reagent, biotin labeled cat hair anaphylactogen and sample to be checked are mixed and reacted, reaction is placed in magnetic field and leaves standstill and remove supernatant, obtains the first solution;
(2) in described first solution, add the anti-mixing of mouse anti human IgE two of alkali phosphatase enzyme mark and react, reaction is placed in magnetic field and leaves standstill and remove supernatant, obtains the second solution;
(3) in described second solution, add luminous substrate and react, surveying luminous intensity;
(4) utilize the calibration object of concentration known to draw luminous intensity standard curve, contrast described typical curve according to the luminous intensity that step (3) obtains, calculate the content of cat hair specific IgE in test serum sample.
In a preferred embodiment of the present invention, the condition of reaction described in step (1) is react 15 minutes at 37 DEG C, and the described standing time is 3 minutes; Also comprise in step (1) and described first solution cleaning fluid is cleaned, be placed in magnetic field and leave standstill and remove supernatant, cleaning repetition 2 times; The condition of reaction described in step (2) is react 15 minutes at 37 DEG C, and the described standing time is 3 minutes; Also comprise in step (2) and described second solution cleaning fluid is cleaned, be placed in magnetic field and leave standstill and remove supernatant, cleaning repetition 2 times; The condition of reaction described in step (3) is react 5 minutes at 37 DEG C, in chemiluminescence analysis/analyzer, measure luminous intensity.
In a preferred embodiment of the present invention, described method is carried out in Full-automatic chemiluminescence apparatus Lumiray series, and described Lumiray series is Lumiray1200, Lumiray1220, Lumiray1260 or Lumiray1280.
In a preferred embodiment of the present invention, the step that described method comprises is:
(1) identify in the agent bin described kit being put into Full-automatic chemiluminescence apparatus;
(2) calibration object is placed in the instrument sample storehouse of described Full-automatic chemiluminescence apparatus, identifies calibration object information, distribute calibration object position;
(3) Quality Control thing or sample to be tested are placed in instrument sample storehouse, editor's Detection Information;
(4) startup optimization program, all calibration objects/Quality Control thing/sample to be tested automatically carries out process and obtains result.
The invention has the beneficial effects as follows: the kit of detection cat hair allergenic specific IgE antibody of the present invention and method, in kit, each reagent component has good stability, and the term of validity can to more than 1 year, and detection sensitivity is high, specific performance good, it is little to make a variation.Through the process optimization of great many of experiments, improve unified technique, and produce in strict accordance with standard production working specification and quality control protocol.User only need carry out standard operation according to operation instructions, just can obtain reliable result.In clinical studies with external import reagent meet correlativity up to more than 90%, and expense be only its 1/2.
Accompanying drawing explanation
In order to be illustrated more clearly in the technical scheme in the embodiment of the present invention, below the accompanying drawing used required in describing embodiment is briefly described, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to these accompanying drawings, wherein:
Fig. 1 is the canonical plotting of the kit detecting cat hair allergenic specific IgE antibody (E1) in embodiments of the invention one;
Fig. 2 is the performance comparison figure of cat hair allergenic specific IgE antibody (E1) detection kit of the present invention and import reagent.
Embodiment
Be clearly and completely described to the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only a part of embodiment of the present invention, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making other embodiments all obtained under creative work prerequisite, belong to the scope of protection of the invention.
Embodiment one:
A kind of kit detecting cat hair allergenic specific IgE antibody (E1) based on nano magnetic particulate Full-automatic chemiluminescence standard measure is provided, comprises following reagent:
(1) biotin labeled cat hair allergen solution, concentration is 0.1-1.0ug/mL;
(2) the nano magnetic microparticle suspending liquid of marked by streptavidin, concentration is 0.1-1.0mg/mL;
(3) the anti-solution of mouse anti human IgE two of alkali phosphatase enzyme mark, concentration is 0.1-2.0ug/mL;
The calibration object of (4) 6 variable concentrations levels: 0IU/ml, 0.35IU/ml, 0.7IU/ml, 3.5IU/ml, 17.5IU/ml, 50IU/ml, 100IU/ml;
The quality-control product of (5) 2 variable concentrations levels: 0.7IU/ml, 17.5IU/ml.
Embodiment two:
The preparation that a kind of nano magnetic particulate Full-automatic chemiluminescence standard measure provided by the invention detects serum cat hair allergenic specific IgE antibody kit comprises the steps:
(1) damping fluid is prepared:
Preparation 0.01M, pH value are the phosphate buffer of 7.4, and 0.1M, pH value are the Tris-HCl damping fluid of 7.4, and 0.05M, pH value are the MES damping fluid of 6.0.
0.01M, pH value are the preparation of the phosphate buffer of 7.4: the beaker of the amount of trying to please 1L, add 800ml deionized water, take 2.56gNa
2hPO
412H
2o, 0.44gNaH
2pO
42H
2o and 9gNaCl, joins in beaker, stirs and makes it fully dissolve, with HCl or NaOH adjust ph to 7.4 ± 0.05, add massfraction be 2% sweet mellow wine, massfraction be the glycerine of 1%, stirring 0.5h to dissolving completely, adding deionized water and being settled to 1L.
0.1M, pH value are the preparation of the Tris-HCl damping fluid of 7.4: the beaker of the amount of trying to please 1L, add 800ml deionized water, take 12.11gTris(trishydroxymethylaminomethane) join in beaker, stirring makes it fully dissolve, with HCl or NaOH adjust ph to 7.4 ± 0.05, add massfraction be 2% BSA, massfraction be the Proclin300 of 0.3%, stirring 0.5h to dissolving completely, adding deionized water and being settled to 1L.
0.05M, pH value are the preparation of the MES damping fluid of 6.0: the beaker of the amount of trying to please 1L, add 800ml deionized water, take 9.76gMES(2-(N-morpholino) ethyl sulfonic acid), 9gNaCl joins in beaker, stirring makes its abundant solution, with HCl or NaOH adjust ph to 6.0 ± 0.05, add massfraction be 1% BSA, massfraction be the Tween-20 of 0.1%, stirring 0.5h to dissolving completely, adding deionized water and being settled to 1L.
(2) biotin labeled cat hair allergen solution is prepared
Being joined with the ratio of 1:20 by cat hair anaphylactogen is dissolved in the biotin of dimethyl formamide, abundant hybrid reaction 30 minutes under room temperature condition, and reacted solution 0.1M, pH value are the phosphate buffer dialysis of 7.4.BCA method measures the concentration of biotin labeled cat hair allergen solution, and adjusts its concentration to 0.1-1.0ug/ml.
(3) the nano magnetic microparticle suspending liquid of marked by streptavidin is prepared
The nano magnetic particulate magnetic separator of marked by streptavidin is precipitated out, with 0.01M, pH value be 7.4 phosphate buffer resuspended, abundant mixing after 15 minutes Magneto separate settle out, supernatant discarded, again with 0.01M, pH value be 7.4 phosphate buffer resuspended, repeated washing like this 5 times, the nano magnetic microparticulate after cleaning is the phosphate buffer of 7.4 in 0.01M, pH value, and concentration is 0.1-1.0mg/ml.
(4) the anti-solution of mouse anti human IgE two of alkali phosphatase enzyme mark is prepared;
By 0.05M, pH value be 6.0 MES damping fluid be diluted to 0.1-2.0ug/ml by anti-for the mouse anti human IgE two of alkali phosphatase enzyme mark.
(5) calibration object is prepared
Get IgE albumen, with the Tris-HCl buffer that 0.1M, pH value are 7.4 become 0,0.35,0.7,3.5,17.5,50, the calibration point of 100IU/ml.
(6) quality-control product is prepared
Get IgE albumen, with the Tris-HCl buffer that 0.1M, pH value are 7.4 become 0.7, the quality-control product of 17.5IU/ml.
(7) kit assembling
The anti-solution of mouse anti human IgE two of the nano magnetic microparticle suspending liquid of biotin labeled cat hair allergen solution, marked by streptavidin, alkali phosphatase enzyme mark, calibration object and quality-control product are assembled into kit.
The reagent component (such as the damping fluid etc. of cleaning fluid, some necessity) do not mentioned in detail in kit of the present invention, the external packing of kit and the independent packaging container etc. of each reagent component all can carry out according to the routine operation in affiliated field, meet relevant industries and specify.
Embodiment three:
Provide the detection method that a kind of nano magnetic particulate Full-automatic chemiluminescence standard measure detects serum cat hair allergenic specific IgE antibody kit, detect and carry out in Full-automatic chemiluminescence apparatus Lumiray series (Lumiray1200, Lumiray1220, Lumiray1260, Lumiray1280 etc.), concrete instrumentation step is as follows:
(1) kit is put into Full-automatic chemiluminescence analysis/analyzer agent bin relevant position, kit information inputs instrument system by barcode scanner, also sets by instrument software kit.
(2) calibration object is placed in instrument sample storehouse.By barcode scanner identification calibration object information, and in instrument system, distribute calibration object position.
(3) Quality Control thing/sample to be tested is placed in instrument sample storehouse, by the corresponding Detection Information of instrument software kit editor.
(4) startup optimization program, all calibration objects/Quality Control thing/sample processing steps to be checked will automatically perform.
A kind of nano magnetic particulate Full-automatic chemiluminescence provided by the invention quantitatively detects serum cat hair allergenic specific IgE antibody kit and can also realize on semi-automatic Chemiluminescence Apparatus, and concrete pattern detection step is as follows:
(1) in detector tube, add the nano magnetic particulate of 40ul test serum or IgE calibration object, 50ul biotin labeled cat hair anaphylactogen, 50ul marked by streptavidin successively, after mixing, 15 minutes are hatched altogether at 37 DEG C, in sample, cat hair specific IgE antibody is captured, and anchors at nano magnetic microparticle surfaces.
(2) under externally-applied magnetic field effect, magnetic particle settles down, and removes supernatant, adds 500ul cleaning fluid, and after removing magnetic field, concussion makes magnetic particle resuspended; Repeat this cleaning step 3 times to remove unconjugated biotin labeled mouse antihuman IgE antibody.
(3) add the anti-solution of mouse anti human IgE two of 100ul alkali phosphatase enzyme mark, mixing, hatches 15 minutes at 37 DEG C.
(4) under externally-applied magnetic field effect, magnetic particle settles down, and removes supernatant, adds 500ul cleaning fluid, and after removing magnetic field, concussion makes magnetic particle resuspended; Repeat this cleaning step 3 times to remove unconjugated enzyme labelled antibody and other impurity.
(5) add 150ul enzyme-catalyzed chemical luminescence substrate, remove magnetic field, fully after mixing, 37 ° of C hatch 5 minutes, measure luminous value.
(6) process of data: by concentration value and the luminous value detected of calibration object, obtain typical curves by four parametrical nonlinearity matchings, the luminous value of sample is substituted into the typical curve shown in Fig. 1 and obtains corresponding concentration value.
Principle of work of the present invention: the nano magnetic particulate of test serum sample and biotin labeled cat hair anaphylactogen, marked by streptavidin is hatched altogether, in test serum sample, cat hair specific IgE antibody is combined with biotin labeled cat hair allergen specificity, simultaneously by biotin-Streptavidin amplification system, in serum sample, cat hair allergenic specific IgE is attached on solid phase carrier nano magnetic particulate; Under externally-applied magnetic field effect, unnecessary biotin labeled cat hair anaphylactogen is removed in Magneto separate washing; The mouse anti human IgE two adding alkali phosphatase enzyme mark resists, and forms the immunocomplex formation that biotin labeled cat hair anaphylactogen-cat hair specific IgE-anti-IgE bis-is anti-; Under externally-applied magnetic field effect, repeatedly unconjugated enzyme labelled antibody and other impurity are removed in Magneto separate washing; Add enzyme-catalyzed chemical luminescence substrate, measure luminous intensity; Use the calibration object typical curve of concentration known, according to luminous intensity reference standard curve, calculate the content of cat hair specific IgE antibody in sample to be tested;
Embodiment four:
The Performance Evaluation of described kit:
1, cat hair allergenic specific IgE antibody (E1) detection kit (nano magnetic particulate Full-automatic chemiluminescence method) analytical performance and stability, specifically see the following form.
2, Fig. 2 is referred to, the performance comparison of cat hair allergenic specific IgE antibody (E1) detection kit (nano magnetic particulate Full-automatic chemiluminescence method) and import reagent.
3, cat hair allergenic specific IgE antibody (E1) detection kit (nano magnetic particulate Full-automatic chemiluminescence method) clinical reference value scope, specifically sees the following form, using bilateral 95 percentile of normal person's pattern detection value as reference value.
| 95% calculates base region (mIU/mL) |
| normal non-allergy is reacted into single anaphylactogen IgE antibody concentration in the serum of people and is less than 0.35 IU/mL. |
| measured value between 0.35 ~ 0.7 IU/mL be this anaphylactogen 1 grade of allergy; |
| measured value between 0.7 ~ 3.5 IU/mL be this anaphylactogen 2 grades of allergy; |
| measured value between 3.5 ~ 17.5 IU/mL be this anaphylactogen 3 grades of allergy; |
| measured value between 17.5 ~ 50 IU/mL be this anaphylactogen 4 grades of allergy; |
| measured value between 50 ~ 100 IU/mL be this anaphylactogen 5 grades of allergy; |
| measured value is greater than 100 IU/mL for this anaphylactogen 6 grades of allergy. |
The invention has the beneficial effects as follows:
One, the present invention is with alkaline phosphatase (AP) for marker enzyme, by chemical reaction labelled antibody, and uses gel chromatography to be separated unreacted enzyme, antibody or antigen, improves the sensitivity of reaction;
Two, with immune magnetic particle for solid phase, with Streptavidin conjugated magnetic microballoon, as general separation agent, not only make immune response more easily mix and be separated, and substantially increasing reaction velocity;
Three, with new chemical luminous substrate AMPPD for substrate, this substrate is wide variety of glow-type substrate, and reaches plateau fast, is conducive to the detection of signal, improves sensitivity and the specificity performance of final kit; To go forward side by side one-step optimization chemiluminescent enhancement system, ensure that the high and good stability of the signal sensitivity of finished product, make a variation little;
Four, the advantage of AMPPD luminous substrate is adopted to be highly sensitive and platform is long for stationary phase.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize description of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical field, be all in like manner included in scope of patent protection of the present invention.
Claims (10)
1. one kind is detected the kit of cat hair allergenic specific IgE antibody, it is characterized in that, the reagent comprised has: calibration object, quality-control product, biotin labeled cat hair allergen solution, the anti-solution of mouse anti human IgE two of alkali phosphatase enzyme mark, the nano magnetic microparticle suspending liquid of marked by streptavidin.
2. kit according to claim 1, it is characterized in that, the concentration of described biotin labeled cat hair allergen solution is 0.1 ~ 1.0ug/ml, the concentration of the nano magnetic microparticle suspending liquid of described marked by streptavidin is 0.1 ~ 1.0mg/ml, and the concentration of the anti-solution of mouse anti human IgE two of described alkali phosphatase enzyme mark is 0.1 ~ 2.0ug/ml.
3. kit according to claim 1, it is characterized in that, the concentration of described calibration object is: 0IU/ml, 0.35IU/ml, 0.7IU/ml, 3.5IU/ml, 17.5IU/ml, 50IU/ml, 100IU/ml, described calibration object to be IgE albumen and 0.1M, pH value be 7.4 Tris-HCl buffer obtain; The concentration of described quality-control product is: 0.7IU/ml, 17.5IU/ml, described quality-control product to be IgE albumen and 0.1M, pH value be 7.4 Tris-HCl buffer obtain.
4. kit according to claim 1, it is characterized in that, described cat hair anaphylactogen be can with the protein bound antigen of cat hair specific IgE in human body, the preparation process of described biotin labeled cat hair anaphylactogen is: joined by 1:20 mol ratio by cat hair anaphylactogen and be dissolved in the biotin of dimethyl formamide, then obtains with the phosphate buffer dialysis that 0.1M, pH value are 7.4.
5. kit according to claim 1, it is characterized in that, the mouse anti human IgE two of described mark resist for can with the antibody of IgE albumen specific bond in human body, the preparation process that the mouse anti human IgE two of described alkali phosphatase enzyme mark is anti-is: the alkaline phosphatase enzyme solutions by concentration being 1.0-5.0mg/mL is anti-in molar ratio for 2-10:1 carries out mixing rear also purifying with mouse anti human IgE two, the mouse anti human IgE two obtaining alkali phosphatase enzyme mark resists, described purifying balances and wash-out with the bicarbonate buffer that pH is 8-9, UV detect and record purifying collection of illustrative plates again, use 0.05M again, pH value be 6.0 MES damping fluid the mouse antihuman IgE antibody of alkali phosphatase enzyme mark is diluted.
6. kit according to claim 1, it is characterized in that, the preparation process of the nano magnetic microparticle suspending liquid of described marked by streptavidin is: precipitated by the nano magnetic particulate magnetic separator of marked by streptavidin, again with 0.01M, pH value be 7.4 phosphate buffer resuspended, mixing Magneto separate precipitation, and repeated washing, the nano magnetic microparticulate after cleaning in 0.01M, pH value be the phosphate buffer of 7.4.
7. provide a kind of nanometer Magnetism particulate immuno chemistry luminescence method quantitatively to detect the method for serum cat hair allergenic specific IgE antibody, it is characterized in that, comprising step is:
(1) magnetic particle reagent, biotin labeled cat hair anaphylactogen and sample to be checked are mixed and reacted, reaction is placed in magnetic field and leaves standstill and remove supernatant, obtains the first solution;
(2) in described first solution, add the anti-mixing of mouse anti human IgE two of alkali phosphatase enzyme mark and react, reaction is placed in magnetic field and leaves standstill and remove supernatant, obtains the second solution;
(3) in described second solution, add luminous substrate and react, surveying luminous intensity;
(4) utilize the calibration object of concentration known to draw luminous intensity standard curve, contrast described typical curve according to the luminous intensity that step (3) obtains, calculate the content of cat hair specific IgE in test serum sample.
8. method according to claim 7, is characterized in that, the condition of reaction described in step (1) is react 15 minutes at 37 DEG C, and the described standing time is 3 minutes; Also comprise in step (1) and described first solution cleaning fluid is cleaned, be placed in magnetic field and leave standstill and remove supernatant, cleaning repetition 2 times; The condition of reaction described in step (2) is react 15 minutes at 37 DEG C, and the described standing time is 3 minutes; Also comprise in step (2) and described second solution cleaning fluid is cleaned, be placed in magnetic field and leave standstill and remove supernatant, cleaning repetition 2 times; The condition of reaction described in step (3) is react 5 minutes at 37 DEG C, in chemiluminescence analysis/analyzer, measure luminous intensity.
9. method according to claim 7, is characterized in that, described method is carried out in Full-automatic chemiluminescence apparatus Lumiray series, and described Lumiray series is Lumiray1200, Lumiray1220, Lumiray1260 or Lumiray1280.
10. method according to claim 9, is characterized in that, the step comprised is:
(1) identify in the agent bin described kit being put into Full-automatic chemiluminescence apparatus;
(2) calibration object is placed in the instrument sample storehouse of described Full-automatic chemiluminescence apparatus, identifies calibration object information, distribute calibration object position;
(3) Quality Control thing or sample to be tested are placed in instrument sample storehouse, editor's Detection Information;
(4) startup optimization program, all calibration objects/Quality Control thing/sample to be tested automatically carries out process and obtains result.
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| CN115541891A (en) * | 2022-11-25 | 2022-12-30 | 济南德亨医学科技有限公司 | Allergen-specific IgE antibody detection kit and preparation method thereof |
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