CN103616520B - Highly sensitive method for detecting food allergen and kit thereof - Google Patents
Highly sensitive method for detecting food allergen and kit thereof Download PDFInfo
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- CN103616520B CN103616520B CN201310645509.7A CN201310645509A CN103616520B CN 103616520 B CN103616520 B CN 103616520B CN 201310645509 A CN201310645509 A CN 201310645509A CN 103616520 B CN103616520 B CN 103616520B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
- G01N33/54333—Modification of conditions of immunological binding reaction, e.g. use of more than one type of particle, use of chemical agents to improve binding, choice of incubation time or application of magnetic field during binding reaction
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4716—Complement proteins, e.g. anaphylatoxin, C3a, C5a
Abstract
The invention discloses a kind of highly sensitive method for detecting food allergen and kit thereof.1) food allergen albumen coupling magnetic bead, described food allergen includes but not limited to aquatic product, milk, eggs, meat, cereals, nut fruits, fruits, greengrocery, bean food allergen protein; 2) typical curve is built: IgG antibody standard items and the food allergen albumen coupling magnetic bead of the anti-food allergen of gradient dilution carry out immune response, then utilize anti-human igg two to resist and carry out signal iodine, carry out linear regression according to mean absorbance values and corresponding concentration, build typical curve; 3) examination food allergen, utilizes the serum of food allergen albumen coupling magnetic bead and autopath to carry out immune response, then utilizes anti-human igg two to resist and carries out signal iodine, then determine the concentration of the kind of food allergen and the antibody of correspondence.The range of sensitivity of the present invention, not higher than 2pg/mL, is applicable to the Allergic skin test of enetically modified food.
Description
Technical field
The present invention relates to food security and irritated detection field, particularly relate to a kind of highly sensitive method for detecting food allergen and kit thereof.
Background technology
Anaphylactia affects the population in the whole world nearly 1/4th, by one of the World Health Organization's three large diseases being classified as 21 century keypoint control.Food hypersenstivity medically also claims allergic reaction, after referring to that body is stimulated by antigen (comprising haptens), produce corresponding antibody or sensitized lymphocyte, after again contacting same antigen, cause body fluid or cell immune response in vivo, cause tissue damage or organism physiology dysfunction thus.It is the disease that a kind of recurrence rate is very high.According to food irritability or allergic reaction network statistics, about have the allergic reaction of 90% to be cause (allergy caused with peanut is the most serious) by foods such as peanut, kernel, fish, shellfish, egg, milk, soybean, fishes and shrimps, mushroom and wheats, the allergy of 10% is caused by other foods of kind more than 170.According to epidemiology survey, the allergy incidence of disease is in the trend risen year by year in recent years.West probably has the people of 20-25% to be subject to the harassment of anaphylactogen according to investigations, and the people of the nearly 5-10% of China is subject to the harassment of anaphylactogen.The crowd of anaphylactogen harassment is received more than 10% in the coastlands such as Shanghai, Zhejiang, Guangdong, and along with the fast development of industry, growth in the living standard, the improvement of diet, the crowd harassed by anaphylactogen increases with the speed of 1.5% every year.The U.S. about has the human hair uncooked food of 2% ~ 2.5% irritated every year, the incidence of disease about 5% ~ 8% of children and infant.According to incompletely statistics, the incidence of disease of China is higher than developed country.At present, along with scientific-technical progress and expanding economy, create many new food resources, and food production, circulation, consumption pattern change, the region of traditional food hypersenstivity is broken, and along with a large amount of appearance of genetically modified crops, many new enetically modified food emerge in multitude, food hypersenstivity is also made to have had more for potential danger.Food hypersenstivity endangers the health of allergic human population in sizable degree, and Protection of consumer food security has become the major issue that food safety management department and the food enterprises face.
Many people, after direct or these anaphylactogens of indirect contact, can suffer from the diseases such as bronchial astehma, upper respiratory tract conjunctivitis, eczema, dermatitis because of irritated, sometimes even produce life-threatening anaphylactic shock.Therefore go out hiding anaphylactogen in the urgent need to examination from numerous food, allow Yi Minzhe avoid the food containing allergen and to treat allergic reaction, get rid of the mistaken diagnosis of similar allergic symptom chronic disease simultaneously.Autopath is due to after the anaphylactoid food allergen albumen of absorption initiation, and in body, basophil and mast cell degranulation cause specific IgG precipitating antibody to enter blood, and this specific IgG antibodies level in blood is improved.This specific IgG antibodies can identify the antigen of its correspondence in vitro, i. e. allergen protein, and combines with it.
For the detection of allergenicity, vivo experiment method can provide the most directly evidence, as the provocative test of double blinding food and skin puncture test, but due to the consideration of the aspects such as safety factor, only carry out in hospital in unavoidable situation, and somewhat expensive, to have a big risk; Ill vitro test method has the advantage of convenience, safety in contrast to this, but there is the shortcoming of poor accuracy.Along with the development of science and technology, following food allergen detects will to accurate, safety, economy, quick, high flux, highly sensitive vitro detection technique direction development.
The kind of food allergen diagnostic reagent is a lot, and the factor affecting reagent quality is also a lot, for guarantee reagent quality needs to develop skill as much as possible index.Sensitivity, as an important technical indicator, has great importance to the evaluation of kit.Because few quantitative analysis thing may to defining morbid state, disorder in screening is significant.The antibody produced in body in food allergen detects is very micro-, and on market, prior art can only reach nanogram level mostly, in detection lower than just becoming more difficult during this concentration, and is difficult to accomplish the end in view.
Summary of the invention
In order to overcome the deficiencies in the prior art, the object of this invention is to provide a kind of highly sensitive method for detecting food allergen and kit thereof.
A kind of highly sensitive method for detecting food allergen, step is as follows:
1) food allergen albumen coupling magnetic bead, described food allergen includes but not limited to aquatic product, milk, eggs, meat, cereals, nut fruits, fruits, greengrocery, bean food allergen protein;
2) typical curve is built: IgG antibody standard items and the food allergen albumen coupling magnetic bead of the anti-food allergen of gradient dilution carry out immune response, then utilize anti-human igg two to resist and carry out signal iodine, carry out linear regression according to mean absorbance values and corresponding concentration, build typical curve;
3) examination food allergen, the serum of food allergen albumen coupling magnetic bead and autopath is utilized to carry out immune response, then utilize that anti-human igg two is anti-carries out signal iodine, then according to the signal obtained and step 2) kind of typical curve determination food allergen of gained and the concentration of corresponding antibody.
Described signal iodine is chromogenic reaction.
Determine in described step 3) that the cut-off value of the kind of food allergen is set as that the value of antibody concentration 1ng/mL is at absorbance corresponding to typical curve.
In described step 1), concrete grammar is as follows:
1.1) centrifuge tube getting a 5mL adds the NHS solution of the phosphate buffer of 3.84mL 50mM pH4.0, the magnetic bead of 1mL 5%, the EDAC solution of 60uL 50mg/mL and 100uL 100mg/mL wherein respectively, centrifuge tube is placed on shaking table, under room temperature condition, activates 30min;
1.2) centrifuge tube is placed on magnetic frame holds magnetic bead, discard clear liquid, and add the phosphate buffer of 5mL 50mM pH6.5 in centrifuge tube; Repeated washing 3 times, adds the phosphate buffer of 4.499 mL 50mM pH6.5 for the last time;
1.3) add the food allergen albumen 500uL of 5ug/mL more wherein, 1% bovine serum albumin 1uL, is placed in centrifuge tube on shaking table, reacts 45min under 40 DEG C of conditions;
1.4) repeat 1.2) washing step;
1.5) add phosphate buffer and the 10uL 1%BSA solution of 4.990mL 50mM pH6.5, under 4 DEG C of conditions, Seal and preservation is stand-by.
In described step 3), concrete grammar is as follows:
3.1) get the centrifuge tube of some 2mL, add the magnetic bead of 300uL coupling food allergen albumen respectively, hold magnetic bead with magnetic frame, discard clear liquid, then add the phosphate buffer of 300uL pH7.4, so washing 3 times.Finally blot;
3.2) in the magnetic bead after washing, add 120uL, containing 20% glycerine, the antibody of gradient dilution, hatches 40min under room temperature condition;
3.3) after hatching end, hold magnetic bead with magnetic frame, discard clear liquid, then add the PBS cleansing solution of 300uL, whirlpool shakes, then holds magnetic bead with magnetic frame, and so washing 5 times, finally blots liquid;
3.4) in the magnetic bead after washing, add two anti-120uL of the horseradish peroxidase-labeled of 100ng/mL, containing 20% glycerine, under room temperature condition, hatch 40min;
3.5) step 3.3 is repeated);
3.6) in the magnetic bead after washing, the TMB nitrite ion of 100uL is added, lucifuge reaction 15min;
3.7), after lucifuge reaction, add the hydrochloric acid solution of 100uL 1moL/L wherein, cessation reaction, and liquid rotating is moved on in ELISA Plate;
3.8) absorbance is read by microplate reader;
3.9) result calculates.
The surface of described magnetic bead has carboxyl, amino or without group, particle size is at 100nm ~ 3um.
Food allergen assay kit, it includes but not limited to wrap one or more magnetic Nano microsphere in the aquatic product of quilt, milk, eggs, meat, cereals, nut fruits, fruits, greengrocery, bean food allergen protein, human IgG titer, horseradish peroxidase-labeled two anti-, TMB, sample diluting liquid, cleansing solution and stop buffer, and the range of sensitivity is not higher than 2pg/mL.
Described kit, is applicable to the Allergic skin test of enetically modified food.
Beneficial effect of the present invention: using magnetic bead as solid phase carrier, the ELISA Plate micropore in replacing ELISA to detect.This solid phase carrier has larger area load amount compared to the ELISA Plate micropore of common ELISA, and stronger inrichment can be had to target substance, a concentrated effect can be played to object by the process of externally-applied magnetic field and washing, finally improve sensitivity that food allergen detects, accuracy and the range of linearity.The range of sensitivity, not higher than 2pg/mL, is applicable to the Allergic skin test of enetically modified food.
Accompanying drawing explanation
Fig. 1 is the result schematic diagram of reagent Linear Experiment in embodiment four;
Fig. 2 is the result schematic diagram of embodiment reagent Linear Experiment.
Embodiment
Below in conjunction with drawings and Examples, the present invention is described further.
Food allergen assay kit of the present invention, its composition comprise wrap quilt aquatic product, milk, eggs, meat, cereals, nut fruits, fruits, greengrocery, bean food allergen protein magnetic Nano microsphere, human IgG titer, horseradish peroxidase-labeled two resist, TMB, sample diluting liquid, cleansing solution and stop buffer.
1, food allergen albumen coupling magnetic bead
1. the centrifuge tube getting a 5mL adds the NHS solution of the phosphate buffer of 3.84mL 50mM pH4.0, the magnetic bead of 1mL 5%, the EDAC solution of 60uL 50mg/mL and 100uL 100mg/mL wherein respectively, centrifuge tube is placed on shaking table, under room temperature condition, activates 30min.
2. centrifuge tube is placed on magnetic frame and holds magnetic bead, discard clear liquid, and add the phosphate buffer of 5mL 50mM pH6.5 in centrifuge tube; Repeated washing like this 3 times, adds the phosphate buffer of 4.499mL 50mM pH6.5 for the last time.
3. add the food allergen albumen 500uL of 5ug/mL more wherein, 1% bovine serum albumin 1uL, is placed in centrifuge tube on shaking table, reacts 45min under 40 DEG C of conditions.
4. washing step is 2. repeated.
5. phosphate buffer and the 10uL 1%BSA solution of 4.990mL 50mM pH6.5 is added, Seal and preservation under 4 DEG C of conditions.Stand-by.
2, the using method of food allergen assay kit
1. get the centrifuge tube of some 2mL, add the magnetic bead of 300uL coupling food allergen albumen respectively, hold magnetic bead with magnetic frame, discard clear liquid, then add the phosphate buffer of 300uL pH7.4, so washing 3 times.Finally blot.
2. in the magnetic bead after washing, 120uL(is added containing 20% glycerine) antibody of gradient dilution, hatch 40min under room temperature condition.
3. after hatching end, hold magnetic bead with magnetic frame, discard clear liquid, then add the PBS cleansing solution of 300uL, whirlpool shakes, then holds magnetic bead with magnetic frame, and so washing 5 times, finally blots liquid.
4. in the magnetic bead after washing, two anti-120uL(of the horseradish peroxidase-labeled of 100ng/mL are added containing 20% glycerine), hatch 40min under room temperature condition.
5. step is repeated 3..
6. in the magnetic bead after washing, the TMB nitrite ion of 100uL is added, lucifuge reaction 15min.
7., after lucifuge reaction, the hydrochloric acid solution of 100uL 1moL/L is added wherein, cessation reaction.And liquid rotating is moved on in ELISA Plate.
8. absorbance is read by microplate reader.
9. result calculates
Get the absorbance values of parallel three times of repeating hole; Carry out linear regression with the mean absorbance values of series standard solution and corresponding concentration, build typical curve; That absorbance that the mean absorbance values of calculation sample repeating hole is greater than cut-off value (absorbance that 1ng/mL is corresponding) calculates the content of food allergen antibody from typical curve, and this content back has answered the degree of allergic reaction of patient.
embodiment one
1 prawn allergen protein coupled bead
1. the centrifuge tube getting a 5mL adds the NHS solution of the phosphate buffer of 3.84mL 50mM pH4.0, the magnetic bead of 1mL 5%, the EDAC solution of 60uL 50mg/mL and 100uL 100mg/mL wherein respectively, centrifuge tube is placed on shaking table, under room temperature condition, activates 30min.
2. centrifuge tube is placed on magnetic frame and holds magnetic bead, discard clear liquid, and add the phosphate buffer of 5mL 50mM pH6.5 in centrifuge tube; Repeated washing like this 3 times, adds the phosphate buffer of 4.499mL 50mM pH6.5 for the last time.
3. the prawn allergen protein 500uL(adding 5ug/mL is more wherein the acidoglycoprotein that a kind of relative molecular mass is about 36Kd to shrimp allergen, belongs to the tropomyosin in muscle.Protein concentration 1.5mg/mL, is dissolved in PBS, and tire >=1:1000-5000.), 1% bovine serum albumin 1uL, is placed in centrifuge tube on shaking table, reacts 45min under 40 DEG C of conditions.
4. washing step is 2. repeated.
5. phosphate buffer and the 10uL 1%BSA solution of 4.990mL 50mM pH6.5 is added, Seal and preservation under 4 DEG C of conditions.Stand-by.
The detection method of 2 prawn anti-allergen antibodies
1. get the centrifuge tube of 3 2mL, add the magnetic bead of 300uL coupling prawn allergen protein respectively, hold magnetic bead with magnetic frame, discard clear liquid, then add the phosphate buffer of 300uL pH7.4, so washing 3 times.Finally blot.
2. in the magnetic bead after washing, add 120uL(containing 20% glycerine) little mouse-anti prawn allergen protein monoclonal antibody (be dissolved in PBS, in pH7.4, protein concentration 1mg/mL, containing stabilizing agents such as BSA, ELISA (1:500), IHC (1:200), IP (1:50), WB (1:300).), hatch 40min under room temperature condition.
3. after hatching end, hold magnetic bead with magnetic frame, discard clear liquid, then add the PBS cleansing solution of 300uL, whirlpool shakes, then holds magnetic bead with magnetic frame, and so washing 5 times, finally blots liquid.
4. in the magnetic bead after washing, two anti-120uL(of the horseradish peroxidase-labeled of 100ng/mL are added containing 20% glycerine), hatch 40min under room temperature condition.
5. step is repeated 3..
6. in the magnetic bead after washing, the TMB nitrite ion of 100uL is added, lucifuge reaction 15min.
7., after lucifuge reaction, the hydrochloric acid solution of 100uL 1moL/L is added wherein, cessation reaction.And liquid rotating is moved on in ELISA Plate.
8. absorbance is read by microplate reader.
9. result calculates
Get the absorbance values of parallel three times of repeating hole; Carry out linear regression with the mean absorbance values of series standard solution and corresponding concentration, build typical curve; That absorbance that the mean absorbance values of calculation sample repeating hole is greater than cut-off value (absorbance that 1ng/mL is corresponding) calculates the content of prawn anti-allergen antibodies from typical curve.Specificity reaches 91.3%.
embodiment two: food allergen Screening tests
Blood is extracted to 35 routine food allergy patients, is prepared serum, carry out examination food allergen.Detect absorbance, calculating antibody concentration, its intermediate value is greater than the judgement of cut-off value (absorbance that 1ng/mL is corresponding) for positive.Its testing result is as shown in table 1 below:
table 1
Testing result shows, adopt kit of the present invention can easy, rapid, sensitive, detect food allergen accurately.
embodiment three: Precision Experiment
By the using method of above-mentioned food allergen assay kit, the sample getting two variable concentrations in sensing range measures, and each sample measures 7 times, and by 7 testing result calculating mean values, standard deviation and the coefficient of variation, result is as shown in table 2 below.
Table 2
embodiment four: reagent Linear Experiment
By the using method of above-mentioned food allergen assay kit, get the sample of 5 kinds of concentration known, each sample measures 3 times, averages, and result is as shown in table 3 below.
Table 3
R can be found out according to Fig. 1
2=0.9872
embodiment five: reagent Linear Experiment
By the using method of above-mentioned food allergen assay kit, get the sample of 5 kinds of concentration known, each sample measures 3 times, averages, and result is as shown in table 4 below.
Table 4
R can be found out according to Fig. 2
2=0.9964
embodiment six: accuracy validation
By the using method of above-mentioned food allergen assay kit, get the standard items with traceability, detect with reagent, detect 7 times, calculating mean value contrasts with target value, and result is as shown in table 5 below.
Table 5
embodiment seven: sensitivity test
By the using method of above-mentioned food allergen assay kit, measure the sample of 7 kinds of different antibodies concentration, each sample surveys 7 times, by calculating the coefficient of variation (CV%), the point that definition starts to be greater than 20% is the sensitivity of this kit, the sensitivity of detection kit of the present invention is 0.5pg/mL, and result is as shown in table 6 below.
Table 6
embodiment eight: stability test
Under 2 ~ 8 DEG C of storage requirement, respectively at 0 month, in April, in August, Dec and 14 months measure the antibody of same concentration, and each sample surveys 5 times, gets average.Result shows, April, August, Dec, and within 14 months, the difference recorded is very little compared with 0 month, and detection kit of the present invention Absorbable organic halogens 1 year under 2 ~ 8 DEG C of storage requirement is described, result is as shown in table 7 below.
Table 7
Behind reagent Kaifeng, in 30 days, measure the absorbance average of the antibody sample of same concentration, maximal value, minimum value, standard deviation and the absorbance coefficient of variation for 30 times, this kit reagent is behind Kaifeng as seen, and Absorbable organic halogens more than one month, result is as shown in table 8.
Table 8
| Absorbance average | Absorbance minimum value | Absorbance maximum | Absorbance standard is poor | Coefficient of variation CV (%) |
| 0.433 | 0.421 | 0.456 | 4.33 | 5.0 |
Claims (4)
1. a highly sensitive method for detecting food allergen, is characterized in that, step is as follows:
1) food allergen albumen coupling magnetic bead, described food allergen includes but not limited to aquatic product, milk, eggs, meat, cereals, nut fruits, fruits, greengrocery, bean food allergen protein;
2) typical curve is built: IgG antibody standard items and the food allergen albumen coupling magnetic bead of the anti-food allergen of gradient dilution carry out immune response, then utilize anti-human igg two to resist and carry out signal iodine, carry out linear regression according to mean absorbance values and corresponding concentration, build typical curve;
3) examination food allergen, the serum of food allergen albumen coupling magnetic bead and autopath is utilized to carry out immune response, then utilize that anti-human igg two is anti-carries out signal iodine, then according to the signal obtained and step 2) kind of typical curve determination food allergen of gained and the concentration of corresponding antibody;
Described signal iodine is chromogenic reaction;
Determine in described step 3) that the cut-off value of the kind of food allergen is set as that the value of antibody concentration 1ng/mL is at absorbance corresponding to typical curve;
In described step 1), concrete grammar is as follows:
1.1) centrifuge tube getting a 5mL adds the NHS solution of the phosphate buffer of 3.84mL 50mM pH4.0, the magnetic bead of 1mL 5%, the EDAC solution of 60uL 50mg/mL and 100uL 100mg/mL wherein respectively, centrifuge tube is placed on shaking table, under room temperature condition, activates 30min;
1.2) centrifuge tube is placed on magnetic frame holds magnetic bead, discard clear liquid, and add the phosphate buffer of 5mL 50mM pH6.5 in centrifuge tube; Repeated washing 3 times, adds the phosphate buffer of 4.499 mL 50mM pH6.5 for the last time;
1.3) add the food allergen albumen 500uL of 5ug/mL more wherein, 1% bovine serum albumin 1uL, is placed in centrifuge tube on shaking table, reacts 45min under 40 DEG C of conditions;
1.4) repeat 1.2) washing step;
1.5) add phosphate buffer and the 10uL 1%BSA solution of 4.990mL 50mM pH6.5, under 4 DEG C of conditions, Seal and preservation is stand-by;
In described step 3), concrete grammar is as follows:
3.1) get the centrifuge tube of some 2mL, add the magnetic bead of 300uL coupling food allergen albumen respectively, hold magnetic bead with magnetic frame, discard clear liquid, then add the phosphate buffer of 300uL pH7.4, so washing 3 times;
Finally blot;
3.2) in the magnetic bead after washing, add 120uL, containing 20% glycerine, the antibody of gradient dilution, hatches 40min under room temperature condition;
3.3) after hatching end, hold magnetic bead with magnetic frame, discard clear liquid, then add the PBS cleansing solution of 300uL, whirlpool shakes, then holds magnetic bead with magnetic frame, and so washing 5 times, finally blots liquid;
3.4) in the magnetic bead after washing, add two anti-120uL of the horseradish peroxidase-labeled of 100ng/mL, containing 20% glycerine, under room temperature condition, hatch 40min;
3.5) step 3.3 is repeated);
3.6) in the magnetic bead after washing, the TMB nitrite ion of 100uL is added, lucifuge reaction 15min;
3.7), after lucifuge reaction, add the hydrochloric acid solution of 100uL 1moL/L wherein, cessation reaction, and liquid rotating is moved on in ELISA Plate;
3.8) absorbance is read by microplate reader;
3.9) result calculates.
2. method according to claim 1, is characterized in that, the surface of described magnetic bead has carboxyl, amino or without group, particle size is at 100nm ~ 3um.
3. the food allergen assay kit of a method according to claim 1, it is characterized in that, it includes but not limited to wrap one or more magnetic Nano microsphere in the aquatic product of quilt, milk, eggs, meat, cereals, nut fruits, fruits, greengrocery, bean food allergen protein, human IgG titer, horseradish peroxidase-labeled two anti-, TMB, sample diluting liquid, cleansing solution and stop buffer, and the range of sensitivity is not higher than 2pg/mL.
4.
kit according to claim 3, is characterized in that, is applicable to the Allergic skin test of enetically modified food.
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| CN103901215B (en) * | 2014-04-11 | 2016-05-11 | 苏州浩欧博生物医药有限公司 | Chemical luminescent analysis reagent kid of a kind of food allergen and preparation method thereof and detection method |
| CN105785008A (en) * | 2014-12-23 | 2016-07-20 | 北京新华联协和药业有限责任公司 | Food intolerance test kit and preparation method thereof |
| CN104820090A (en) * | 2015-05-13 | 2015-08-05 | 杭州傲敏生物科技有限公司 | Detecting kit for specific IgE antibody of food allergen as well as preparation and detecting methods of detecting kit |
| CN112505026A (en) * | 2020-11-23 | 2021-03-16 | 深圳大学 | Visual gel detection device and method for soybean allergen |
| CN112710596A (en) * | 2020-11-30 | 2021-04-27 | 浙江正熙生物医药有限公司 | Method for qualitative/quantitative detection of target antibody concentration using flow cytometer |
| CN116242805B (en) * | 2023-02-03 | 2024-01-23 | 温州泛波激光有限公司 | Laser detection method, laser detection device, and computer storage medium |
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| CN101865924A (en) * | 2010-06-26 | 2010-10-20 | 上海交通大学 | Method for detecting fumonisins based on immuno-magnetic bead combined enzyme-linked immunosorbent assay |
| CN102253199A (en) * | 2011-06-14 | 2011-11-23 | 浙江大学 | Method for detecting allergen-specific antibody in serum |
| CN102331492A (en) * | 2011-06-14 | 2012-01-25 | 浙江大学 | Method for detecting mite allergen specific antibody in blood serum |
| CN103323603A (en) * | 2013-06-07 | 2013-09-25 | 博奥生物有限公司 | Protein covalent coupling method on surface of magnetic beads |
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