CN109283336A - The ELISA kit of anaphylactogen is passed based on IgD antibody capture gas - Google Patents
The ELISA kit of anaphylactogen is passed based on IgD antibody capture gas Download PDFInfo
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Abstract
The ELISA kit provided by the invention that anaphylactogen is passed based on IgD antibody capture gas is belonged to gas and passes anaphylactogen detection technique field.The kit includes following reagent, is coated with the detection plate of anti-human IgD antibody；Washing buffer；Sample diluting liquid；Standard items；The gas of biotin labeling passes the anti-human IgD antibody preparation of allergen formulations and biotin labeling；The avidin solution of enzyme label；Substrate developing solution；Terminate liquid.The present invention selects IgD to detect gas and pass anaphylactogen, and is the ELISA kit based on prize law preparation, has the characteristics of high sensitivity, favorable reproducibility, and being capable of the large batch of sample of single treatment.Embodiment proves that kit detection sensitivity provided by the invention reaches 7.81ng/ml.
Description
Technical field
The present invention relates to the technical fields of Allergic skin test kit, and in particular to is transmitted through based on IgD antibody capture gas quick
Former kit.
Background technique
Anaphylactia disease incidence accounts for 30% of total world population or more, is classified as the four big of 21 century by the World Health Organization
One of noninfectious disease.The disease incidence of anaphylactia significantly rises in recent years, becomes the public health problem of global concern,
Guo Jia et al. is to anaphylactia epidemiological survey the results show that there is 22% people to suffer from anaphylaxis disease in 1,200,000,000 total populations
Disease, such as allergic rhinitis, asthma, conjunctivitis, eczema, food hypersenstivity, drug allergy and serious allergic reaction etc., serious shadow
Ring people's lives quality and life security.
Gas passes anaphylactogen such as pollen particles, fungal spore, dust mite, grain flour, animal hair and urine, feather of birds etc.
The major incentive of anaphylactia, wherein pollen is the main source that gas passes anaphylactogen, results in 10%~20% anaphylaxis
The generation of disease.Different pollen has different protein ingredients, and antigenicity is also inconsistent, and there may be friendships between different pollen
Fork reactivity.Dust mite mainly has dermatophagoides pteronyssinus, dust mite, tropical flukeless mite etc., causes allergic rhinitis and bronchial asthma
Important anaphylactogen.Cat and dog are the sources of common zoo-anaphylactogen, mouse, cavy, horse, rabbit in some specific working environments
Also it can become important allergenic source, the hair of the above animal, epithelium, scurf, urine, saliva can have very strong sensitization
Property.Such disease is increasing in recent years, becomes common disease, frequently-occurring disease, is that China's health and economic development field need to solve
Significant problem
Based on the definition of anaphylactia " being one group of disease mediated by mast cell/basophilic granulocyte ", for
For susceptible person through sensitization, any a large amount of mast cells/basophilic granulocyte activation substance that can induce can induce allergy
Reaction.In these substances, the allergic reaction that IgE is mediated is undoubtedly most study, understands most deep one, the clinic in more than 40 years
Practice display, the allergic reaction that IgE is mediated are the most important hypotypes in allergic reaction.Traditionally caused by gas transmissibility anaphylactogen
Allergic reaction is also the allergic reaction mediated by IgE, for example, the Chinese patent of Publication No. CN105403692A discloses detection
The kit and method of house dust mite allergen specific IgE antibody, the Chinese patent of Publication No. CN103983789A disclose
A kind of test strips for specific dust mite class anaphylactogen IgE screening.As it can be seen that the research that people pass anaphylactogen to gas at present stops
In the allergic reaction mediated by IgE, this will lead to the generation that cannot timely and accurately determine anaphylactia.
Summary of the invention
In view of this, it is an object of the invention to pass the kit of anaphylactogen, kit tool based on IgD antibody test gas
There are stronger specificity and higher sensitivity.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides the ELISA kits that anaphylactogen is passed based on IgD antibody capture gas, including following components:
It is coated with the detection plate of anti-human IgD monoclonal antibody；
Washing buffer；
Sample diluting liquid；
Standard items；
The gas of biotin labeling passes the anti-human IgD antibody preparation of allergen formulations and biotin labeling；
The avidin solution of enzyme label；
Substrate developing solution；
Terminate liquid.
Preferably, the peridium concentration of the anti-human IgD monoclonal antibody is 0.1~2000 μ g/ml.
Preferably, the washing buffer is PBS buffer solution.
Preferably, the biotin labeling gas passes the matter of the anti-human IgD antibody preparation of allergen formulations and biotin labeling
Amount concentration stands alone as 0.01~8000 μ g/ml.
Preferably, the anti-human IgD antibody preparation of the biotin labeling is the preparation of the only anti-human IgD of biotin labeling.
Preferably, the sample diluting liquid is PBS buffer solution.
Preferably, the component of the PBS buffer solution comprises the following components in parts by weight: 8.0 parts of NaCl, 0.20 part of KCl,
1.16 part Na2HPO4, 0.20 part of KH2PO4With 1000 parts of water；The pH value of the PBS buffer solution is 7.0~7.6.
Preferably, the hydrogen that the sulfuric acid solution or mass concentration that the terminate liquid is 0.5~2.0mol/L are 30%~40%
Sodium hydroxide solution.
Preferably, the avidin solution of the enzyme label is 1~5000 times of working solution when using.
Preferably, it includes dermatophagoides pteronyssinus, powder that the gas of the biotin labeling, which passes the type that gas passes anaphylactogen in allergen formulations,
Dust mite, tropical flukeless mite, Groton bug, American cockroach, dog epithelium, cat epithelium, argy wormwood, artemisia annua, Sievers wormwood, humulus grass, three are split
Leaf artemisiifolia, timothy grass, birch, poplar, willow, elm, Chinese wax, oriental plane tree, pine tree, cypress, soft leaf thorn certain herbaceous plants with big flowers, is handed at short artemisiifolia
Neurospora, aspergillus fumigatus, penicillium notatum, bread mold, aspergillus niger, grain flour, feather of birds, bee venom, latex or lily.
The ELISA kit provided by the invention that anaphylactogen is passed based on IgD antibody capture gas, including following reagent, coating
There is the detection plate of anti-human IgD monoclonal antibody；Washing buffer；Sample diluting liquid；Standard items；Biotin labeling gas passes anaphylactogen
The anti-human IgD antibody preparation of preparation and biotin labeling；The avidin solution of enzyme label；Substrate developing solution；Terminate liquid.The present invention
Gas biography anaphylactogen is detected using IgD monoclonal antibody is selected for the first time, and is the ELISA kit based on prize law preparation,
With detection high specificity, the characteristics of high sensitivity, favorable reproducibility, and being capable of the large batch of sample of single treatment.Implement illustration
Bright kit detection sensitivity provided by the invention reaches 7.81ng/ml.
Further, the ELISA kit provided by the invention that anaphylactogen is passed based on IgD antibody capture gas, standard items are
People's IgD solution.The gas of biotin labeling, which passes allergen protein, can detecte any air-borne transmission in the liquid such as serum, blood plasma
Allergen-induced body generate allergen specificity IgD, have detection gas pass anaphylactogen type it is more, have a wide range of application
Feature.
Detailed description of the invention
Fig. 1 is that kit of the present invention makes and uses flow chart；
Fig. 2 is the standard curve that IgD antibody capture gas passes anaphylactogen in embodiment 1.
Specific embodiment
The present invention provides the ELISA kits that anaphylactogen is passed based on IgD antibody capture gas, including following reagent:
It is coated with the detection plate of anti-human IgD monoclonal antibody；
Washing buffer；
Sample diluting liquid；
Standard items；
The gas of biotin labeling passes the anti-human IgD antibody preparation of allergen formulations and biotin labeling；
The avidin solution of enzyme label；
Substrate developing solution；
Terminate liquid.
It includes being coated with the detection plate of anti-human IgD monoclonal antibody that the present invention, which provides ELISA kit,.The anti-human IgD
The peridium concentration of monoclonal antibody is preferably 0.1~2000 μ g/ml, more preferably 1~100 μ g/ml.In the present invention, the inspection
The type of drafting board is preferably transparent polystyrene board.The transparent polystyrene board is used for quantitative detection.
ELISA kit provided by the invention includes washing buffer.In the present invention, the washing buffer is preferably
PBS buffer solution.The solute of the PBS buffer solution preferably includes component 8.0 parts of NaCl, the 0.20 part of KCl of following parts by weight,
1.16 part Na2HPO4With 0.20 part of KH2PO4With 1000 parts of water；The pH value of the PBS buffer solution is 7.0~7.6.
ELISA kit provided by the invention includes that biotin labeling gas passes the anti-human of allergen formulations and biotin labeling
IgD antibody preparation.The quality that the biotin labeling gas passes the anti-human IgD antibody preparation of allergen formulations and biotin labeling is dense
Degree is preferably 0.01~8000 μ g/ml, more preferably 0.1~1000 μ g/ml, most preferably 1~100 μ g/ml.In the present invention,
It includes any kind of mistake sucked through respiratory tract that the biotin labeling gas, which passes gas in allergen formulations and passes the type of anaphylactogen,
Quick original, such as dermatophagoides pteronyssinus, dust mite, tropical flukeless mite, Groton bug, American cockroach, dog epithelium, cat epithelium, argy wormwood, chrysanthemum
Wormwood artemisia, Sievers wormwood, humulus grass, Ambrosia trifida, short artemisiifolia, timothy grass, birch, poplar, willow, elm, Chinese wax, oriental plane tree, pine
Tree, cypress, soft leaf thorn certain herbaceous plants with big flowers, Alternaria, aspergillus fumigatus, penicillium notatum, bread mold, aspergillus niger, grain flour, feather of birds, bee venom,
Latex or lily.
In the present invention, the ELISA kit includes sample diluting liquid.The sample diluting liquid is that pH value is 7.0~7.6
PBS buffer solution.The component of the PBS buffer solution includes following parts by weight of component: 8.0 parts of NaCl, 0.20 part of KCl, and 1.16 parts
Na2HPO4, 0.20 part of KH2PO4With 1000 parts of water.
ELISA kit provided by the invention includes standard items.The standard items behaviour IgD albumen.
ELISA kit provided by the invention includes terminate liquid.The terminate liquid is the sulfuric acid solution of 0.5~2.0mol/L
Or the sodium hydroxide solution that mass concentration is 30%~40%.
ELISA kit provided by the invention includes the avidin solution of enzyme label.The avidin solution of the enzyme label
1~5000 times of working solution when to use.The source of the avidin solution of the enzyme label is not particularly limited, using this field
The avidin solution of the label of enzyme known to technical staff.
ELISA kit provided by the invention includes substrate developing solution.In the present invention, the additive amount of the substrate developing solution
The hole preferably 0.02~0.2ml/, the hole more preferably 0.03~0.16ml/.The substrate developing solution is preferably tetramethyl benzidine
(TMB) solution or 2,2- hydrazine-bis- -3- ethyl benzo thiazole phenanthroline -6- sulfonic acid (ABTS) solution.The matter of the tetramethyl benzidine
Measuring concentration is preferably 0.02~0.05%；The 2,2- hydrazine-bis- -3- ethyl benzo thiazole phenanthroline -6- sulfonic acid mass concentration is excellent
It is selected as 0.05~0.1mg/ml.
In the present invention, the preparation method of the ELISA kit of anaphylactogen is passed based on IgD antibody capture gas, preferably includes to wrap
The preparation method for the detection plate for being had anti-human IgD monoclonal antibody.
In the present invention, the preparation method of the detection plate for being coated with anti-human IgD antibody preferably includes following steps:
1) anti-human IgD monoclonal antibody is diluted with coating buffer, obtains coating buffer；
2) coating buffer that the step 1) obtains is added in the reacting hole of detection plate, 4 DEG C overnight；
3) next day adds confining liquid, washes again after incubation in step 2) described in once washing after overnight detection plate hole
It washs, obtains the detection plate for being coated with anti-human IgD monoclonal antibody.
The present invention dilutes anti-human IgD monoclonal antibody with coating buffer, obtains coating buffer.In the present invention, the dilution
Method be not particularly limited, using diluted technical solution well-known to those skilled in the art.It is described in the present invention
Being coated with buffer is preferably carbonate buffer solution.The concentration of the carbonate buffer solution is preferably 0.01~0.2mol/L.It is described
The pH value of carbonate buffer solution is preferably 8.5~9.5.
In the present invention, the concentration of anti-human IgD monoclonal antibody is preferably 0.1~2000 μ g/ml in the coating buffer, more excellent
It is selected as 1~100 μ g/ml.
After obtaining coating buffer, the coating buffer is added in the reacting hole of detection plate by the present invention, and 4 DEG C overnight.The present invention
In, the volume of coating buffer is preferably 0.02~0.2ml coating buffer in each reacting hole.
In the present invention, described 4 DEG C preferably carry out under conditions of standing overnight, and it is firm to be conducive to anti-human IgD monoclonal antibody
Be coated in detection plate.
Next day after overnight detection plate hole described in once washing, adds confining liquid, washs again after incubation.The present invention couple
There is no limit using the technical solution of washing well-known to those skilled in the art for the method for the once washing.This hair
In bright, the additive amount of the washing buffer is preferably at least the volume of coating buffer；The number of the once washing is preferably 2~
6 times；The time of the single wash is preferably every time 0~5min (similarly hereinafter).
After the once washing, confining liquid is added in the detection plate after once washing by the present invention, is washed again after incubation.This
In invention, the confining liquid is preferably the PBS buffer solution for the balf serum albumin that mass concentration is 1%~5%.The closing
The additive amount of liquid is preferably at least the volume of coating buffer.In the present invention, the time of the incubation is preferably 10~200min.It is described
The temperature of incubation is preferably 10~40 DEG C, and more preferably 15~35 DEG C.
After the incubation, the present invention will test plate and be washed again, in the present invention, the method washed again with
The method of once washing described in above-mentioned technical proposal is identical, and details are not described herein.
It is described wash again after, after the present invention preferably drains the moisture in detection plate naturally, with vacuum bag sealing detection plate,
Obtain the detection plate for being coated with anti-human IgD antibody.
The detection method of the ELISA kit of anaphylactogen is passed based on IgD antibody capture gas, comprising the following steps:
1) sample to be tested, standard items, negative controls are added to the detection plate for being coated with anti-human IgD monoclonal antibody
In, it carries out carrying out first time washing after being once incubated for；
2) gas that biotin labeling is added in the detection plate into the step 1) after washing passes allergen formulations, secondary to incubate
It carries out washing for second after educating；
3) avidin solution that enzyme marks is added in the detection plate into the step 2) after washing, is carried out after being incubated for three times
Third time is washed；
4) to substrate developing solution will be added in the detection plate after washing in the step 3), after four times are incubated for, to detection plate
Middle addition terminate liquid, the detection plate to be developed the color；
5) the color developing detection plate obtained according to the step 4), obtains quantitative result.
Sample to be tested is added in the detection plate for being coated with anti-human IgD monoclonal antibody by the present invention, is once incubated for laggard
Row washs for the first time.In the present invention, the sample to be tested includes the liquid containing IgD such as serum or blood plasma.The serum or blood
The dilution of the liquid such as slurry is 1~32000 times.
In the present invention, in order to keep testing result accurately credible, blank control wells, negative control hole, standard are preferably also set up
Sample wells, and guarantee that the amount of liquid of each reacting hole addition is consistent.Sample diluting liquid is preferably added in the blank well；The feminine gender
The liquid samples such as healthy serum or the blood plasma without allergy are preferably added in control wells；The standard sample wells preferably adds people's IgD egg
It is white.
In the present invention, the temperature being once incubated for is preferably 10~40 DEG C, and more preferably 15~35 DEG C.It is described once to incubate
The time educated is preferably 10~200min.
In the present invention, the first time wash temperature is preferably 10~40 DEG C, and more preferably 15~35 DEG C；The washing is slow
The additive amount of fliud flushing is preferably at least the volume of coating buffer；The number of the once washing is preferably 2~6 times；The single is washed
The incubation time washed is preferably every time 0~5min (similarly hereinafter).
The gas that biotin labeling is added into the detection plate after first time washing passes allergen formulations or biotin mark
The anti-human IgD antibody preparation of note, carries out washing for second after secondary incubation.In the present invention, the gas of the biotin labeling is transmitted through
The additive amount of quick original preparation is preferably the hole 0.02~0.2ml/, the hole more preferably 0.03~0.16ml/.
In the present invention, the temperature of the secondary incubation is preferably 10~40 DEG C, and more preferably 15~35 DEG C.It is described secondary to incubate
The time educated is preferably 10~200min.
In the present invention, the method for second of washing is not particularly limited, and use is well-known to those skilled in the art
Washing methods.
After described second is washed, the Avidin of enzyme label or the chain of enzyme label are added into obtained detection plate by the present invention
Mould avidin solution carries out third time washing after being incubated for three times.In the present invention, the Avidin of the enzyme label is using preceding progress
Dilution, preferably 1~5000 times of the diluted multiple.In the present invention, the Avidin of the enzyme label or Streptavidin are molten
The additive amount of liquid is preferably the hole 0.02~0.2ml/, the hole more preferably 0.03~0.16ml/.
In the present invention, the temperature being incubated for three times is preferably 10~40 DEG C, and more preferably 15~35 DEG C.It is described to incubate three times
The time educated is preferably 10~200min.
In the present invention, the method for the third time washing is not particularly limited, and use is well-known to those skilled in the art
Washing methods.
After washing three times, substrate developing solution is added into obtained detection plate by the present invention, after four times are incubated for, into detection plate
Add terminate liquid, the detection plate to be developed the color.
In the present invention, the additive amount of the substrate developing solution is preferably the hole 0.02~0.2ml/, more preferably 0.03~
The hole 0.16ml/.
In the present invention, the temperature of four incubations is preferably 10~40 DEG C, and more preferably 15~35 DEG C.The incubation
Time is preferably 1~45min, more preferably 10~30min.
In the present invention, the sulfuric acid solution or mass concentration that the concentration of the terminate liquid is 0.5~2.0mol/L be 30%~
40% sodium hydroxide solution.The additive amount of the terminate liquid is preferably the hole 0.02~0.2ml/, more preferably 0.03~
The hole 0.16ml/.
The detection plate of the colour developing obtained described in observation or detection, obtains quantitative result.
In the present invention, the quantitative result is judged by the OD value of solution in detection detection plate reacting hole, with blank
The OD value in each hole is surveyed after control wells zeroing, and carries out quantitative detection based on standard curve.It is described detection preferably when with
ABTS colour developing, then Detection wavelength is set as 405nm；When TMB colour developing, then Detection wavelength is set as 450nm.
The kit provided by the invention for passing anaphylactogen based on IgD detection gas is carried out specifically below with reference to embodiment
It is bright, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
Coating: it is 2.0 μ g/ that specific anti-human IgD monoclonal antibody, which is diluted to protein content, with carbonate buffer solution
The coating buffer of ml.0.05ml coating buffer is separately added into each reacting hole of polystyrene board, 4 DEG C overnight.Next day discards in hole
0.35ml washing buffer is added in solution, every hole, washs 3 times, is incubated for 1.5min (referred to as washing, similarly hereinafter) every time；
Closing: the PBS buffer solution that 0.05ml contains 2% balf serum albumin is separately added into each reacting hole, pH value:
7.2 (confining liquids), 15~35 DEG C of incubation 120min, discard solution in hole, washing is same as above；
Be added gradient dilution people IgD albumen (standard sample wells, concentration be shown in Table 1), it is PBS buffer solution (blank control wells), non-
Allergy volunteers sera (negative control hole) and sample to be tested, that is, suspicious dust mite and mugwort pollen allergic patients sera (sample well,
Serum dilutes 5 times with PBS buffer solution, is shown in Table 2)；15~35 DEG C of incubation 60min, are washed out；
Add detection antibody: the anti-human IgD Dan Ke of standard sample wells and the every hole addition 0.05ml biotin labeling of blank control wells
Grand antibody (concentration 2000ng/ml), negative control hole and the every hole of sample well are separately added into the family dirt of corresponding biotin labeling
The Sievers wormwood pollen allergens crude extract of mite allergen crude extract and biotin labeling, 15~35 DEG C of incubation 60min, is then washed
It washs；
The avidin solution of enzyme label: the Avidin that every hole is separately added into 0.05ml horseradish peroxidase-labeled is molten
Liquid, 15~35 DEG C of incubation 30min, is washed out；
Add substrate solution to develop the color: every hole is separately added into the TMB solution 0.05ml of Fresh, and 15~35 DEG C are protected from light incubation
25min；
Terminate reaction: every hole is separately added into 2mol/L sulfuric acid solution 0.05ml；
Detection OD value: ELISA Plate is placed in microplate reader, the absorbance value (OD at Detection wavelength 450nm450), with blank
The OD value (table 1) in each hole is surveyed after control wells zeroing, and draws standard curve.The each Kong Jun of this experiment does multiple holes, and experiment repeats 3
It is secondary.
1 people's IgD antibody capture gas of table passes the standard concentration and its OD value of the ELISA kit of anaphylactogen
| Serial number | Standard items (ng/ml) | OD450It is worth (mean value) |
| 1 | 1000 | 1.3671 |
| 2 | 500 | 1.2386 |
| 3 | 250 | 0.8659 |
| 4 | 125 | 0.4987 |
| 5 | 62.5 | 0.2484 |
| 6 | 31.25 | 0.1988 |
| 7 | 15.63 | 0.1761 |
| 8 | 7.81 | 0.1606 |
| 9 | 3.91 | 0.1582 |
| 10 | 0 | 0.1577 |
Standard curve is drawn according to the concentration of 1,2,3,4,5,6,7,8 and 10 IgD standard items and OD Value Data, such as Fig. 2,
Four parameter Logistic Fitting curve equations: Y=(A-D)/[1+ (X/C) are obtained according to standard curveB]+D；Wherein A=
1.43680；B=-1.94693；C=218.931；D=0.16209；R2=0.99940.
Available by embodiment 1: (1) it is 7.81 that IgD antibody capture gas, which passes the ELISA kit detection range of anaphylactogen,
~1000ng/ml；(2) detection sensitivity of IgD antibody is 7.81ng/ml；(3) gas of quantitative detection sample to be tested passes anaphylactogen
The concentration of specificity IgD simultaneously judges: if the concentration that the gas of sample to be tested passes anaphylactogen specificity IgD is higher than negative control
The concentration in hole can be judged to passing anaphylactogen allergy to the gas；If the gas of sample to be tested passes the concentration of anaphylactogen specificity IgD
Concentration less than or equal to negative control hole can be judged to passing the gas anaphylactogen not allergy.
2 people's IgD antibody capture gas of table passes the case where anaphylactogen specificity IgD antibody
As seen from the above embodiment, the ELISA kit provided by the invention that anaphylactogen is passed based on IgD antibody capture gas,
IgD is selected to detect gas and pass anaphylactogen, and the ELISA kit based on prize law preparation, has detection sensitivity strong, accurately
The characteristics of degree is high, favorable reproducibility, and being capable of the large batch of sample of single treatment.Embodiment proves kit inspection provided by the invention
It surveys sensitivity and reaches 7.81ng/ml.The type for passing anaphylactogen with detection gas is more, the characteristics of having a wide range of application.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. passing the ELISA kit of anaphylactogen based on IgD antibody capture gas, which is characterized in that including following components:
It is coated with the detection plate of anti-human IgD monoclonal antibody；
Washing buffer；
Sample diluting liquid；
Standard items；
The gas of biotin labeling passes the anti-human IgD antibody preparation of allergen formulations and biotin labeling；
The avidin solution of enzyme label；
Substrate developing solution；
Terminate liquid.
2. kit according to claim 1, which is characterized in that the peridium concentration of the anti-IgD monoclonal antibody is 0.1
~2000 μ g/ml.
3. kit according to claim 1, which is characterized in that the washing buffer is PBS buffer solution.
4. kit according to claim 1, which is characterized in that the biotin labeling gas passes allergen formulations and biology
The mass concentration of the anti-human IgD antibody preparation of element label stands alone as 0.01~8000 μ g/ml.
5. kit according to claim 1, which is characterized in that the anti-human IgD antibody preparation of the biotin labeling is
The preparation of the only anti-human IgD antibody of biotin labeling.
6. kit according to claim 1, which is characterized in that the sample diluting liquid is PBS buffer solution.
7. the kit according to claim 3 or 6, which is characterized in that the PBS buffer solution includes the group of following parts by weight
Point: 8.0 parts of NaCl, 0.20 part of KCl, 1.16 parts of Na2HPO4, 0.20 part of KH2PO4With 1000 parts of water；The pH of the PBS buffer solution
Value is 7.0~7.6.
8. kit according to claim 1, which is characterized in that the terminate liquid is that the sulfuric acid of 0.5~2.0mol/L is molten
The sodium hydroxide solution that liquid or mass concentration are 30%~40%.
9. kit according to claim 1, which is characterized in that the concentration of the avidin solution of the enzyme label is to use
When 1~5000 times of working solution.
10. kit according to claim 1, which is characterized in that the biotin labeling gas passes gas in allergen formulations
The type for passing anaphylactogen includes dermatophagoides pteronyssinus, dust mite, tropical flukeless mite, Groton bug, American cockroach, dog epithelium, cat epithelium, Chinese mugwort
Wormwood artemisia, artemisia annua, Sievers wormwood, humulus grass, Ambrosia trifida, short artemisiifolia, timothy grass, birch, poplar, willow, elm, Chinese wax, France
Chinese parasol tree, pine tree, cypress, soft leaf thorn certain herbaceous plants with big flowers, Alternaria, aspergillus fumigatus, penicillium notatum, bread mold, aspergillus niger, bee venom, grain flour,
Feather of birds, latex or lily.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811106725.3A CN109283336A (en) | 2018-09-21 | 2018-09-21 | The ELISA kit of anaphylactogen is passed based on IgD antibody capture gas |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811106725.3A CN109283336A (en) | 2018-09-21 | 2018-09-21 | The ELISA kit of anaphylactogen is passed based on IgD antibody capture gas |
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| CN112710837A (en) * | 2020-11-03 | 2021-04-27 | 浙江大学 | Method for quantifying nsLTP allergen in artemisia pollen |
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