CN1614420A - Method for inspecting dermatophgoides allergen on air conditioner screen and in dust in dust vacuum cleaner - Google Patents
Method for inspecting dermatophgoides allergen on air conditioner screen and in dust in dust vacuum cleaner Download PDFInfo
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- CN1614420A CN1614420A CN 200410051395 CN200410051395A CN1614420A CN 1614420 A CN1614420 A CN 1614420A CN 200410051395 CN200410051395 CN 200410051395 CN 200410051395 A CN200410051395 A CN 200410051395A CN 1614420 A CN1614420 A CN 1614420A
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Abstract
A method for detecting mite idiotoxin in dust includes collecting dust sample through PBS; using Der pII, Der fII singleresistance nano magnetic encapsulated particle and labelled Der pII, Der fII multiresistance nanomagnetic encapsulated particle separately to form solid phase encapsulated composite after inculbation, adding AMPPD substrate as chemical luminescent agent to produce mesometric body AMPD; emitting photon by AMPD after it being decomposed, recording light intensity and calculating out idiotoxin concentration in the sample.
Description
Technical field
The present invention relates to the method that dust mite allergen detects, specifically a kind of method of using magnetic nanoparticle enzyme-linked immunoassay technology for detection Air Conditioning screen pack dust and suction cleaner to collect dust mite allergen Der pII and Der fII content in the dust.
Background technology
Anaphylactia (as asthma, allergic rhinitis, allergic dermatitis etc.) is common disease, a frequently-occurring disease clinically, points out that according to WHO/IAACI in 2000 report pathogenesis of asthma rate and mortality ratio were in rising trend in the last few years.Part city, the whole nation is 3.3%-5.1% to teenager's asthma epidemiology statistics (ISSAC investigation) asthma prevalence, far beyond (about 1%) rising before 10 years; The whole nation has at least 10,000,000 above children to suffer from asthma at present.
Anaphylactia is owing to various contacts or suck allergen and cause that the allergen major part is a protein, also can be the material of polypeptide or carbohydrate and synthetic such as medicine etc.Modal allergen is the dirt mite.The dirt mite is one of strong allergen, and the general allergic disease that can cause comprises allergic asthma, allergia nose bulbar conjunctiva inflammation, AE/dermatitis, allergic urticaria etc.Mite property allergic effect disease accounts for about 70% of various allergic diseases.Most of area in the China and the world distributes the most widely, studies two kinds of the most deep dirt mites that cause the general allergic disease and comprises dermatophagoides pteronyssinus and dust mite.Their hobbies are multiplied in warm, wet environment, find that through seminar's investigation of the school of life and health sciences Liu Zhigang professor of Shenzhen University leader the Air Conditioning filter screen also is one of dwelling place of their hobbies.Air-conditioning just has many cotton-shaped attachments after using a period of time on the air filter screen, these attachments derive from human body exuviae bits, cotton fibre, sweep health, pollen, take off change one's clothes, suspension that smoking etc. produces is aggregative.Dust mite and dermatophagoides pteronyssinus are liked being grown in this warm, moist and have in the environment of food such as scurf.If scavenging air filter screen irregularly or never, so, air-conditioning one is opened, sensitizers such as dirt mite on the air filter screen and dirt mite secretion excreta will be produced by boasting, be deposited in the respiratory tract after being attached on the human body skin or sucking, thereby produce pruitus, have a stuffy nose, sneeze or anaphylactia such as asthma.People are more and more in sealing air-conditioning office work, living and studying time, thereby dust mite allergen also can become one of indoor main sensitizing agent in the room conditioning air filter screen dust.
Indoor environment generally make a general reference people's living quarter, work, work office and indoor place, carry out the public place of other indoor activities etc.Indoor environment has become modern humans's life, worked and has carried out other movable main place.In recent years studying data at home and abroad shows that the time of about 70%-90% city dweller's every day is spent in various indoor environments, therefore, the quality of indoor air environment quality, increasing to health effects.
Indoor biotic pollution mainly contains biological anaphylactogen and pathogenic microorganism etc.Biological anaphylactogen mainly is indoor air microbes such as dirt mite, fungi, bacterium and pollen, brings out human body and anaphylactic allergic reaction's disease occurs, as allergic rhinitis, asthma etc.Wherein the dirt mite is one of topmost anaphylactogen.
In order to prevent and to control the biological anaphylactogen of house dust mite and pollute, the Liu Zhigang professor of life science institute of Shenzhen University leader's seminar has carried out the development of New-generation healthy household electrical appliances, and what mainly comprise the air-conditioning of control dirt mite and band HEPA filter screen removes the mite suction cleaner.
In order to monitor air-conditioning and indoor other local dirt mite pollution situation, and can quantitative response come out, the present invention uses magnetic nanoparticle enzyme-linked immunoassay technology for detection Air Conditioning filter screen dust and suction cleaner to collect the content of dust mite allergen Der p II and Der fII in the dust.
Dirt mite II group anaphylactogen (Der p II and Der fII) is that molecular weight is 15KDa, and the albumen of n terminal amino acid sequence is much at one arranged, and also is one of topmost dust mite allergen of draining by the intestines and stomach of acarid.
The present invention is by magnetic nanoparticle enzyme-linked immunoassay technology, and the house dust mite pollution situation was monitored when the content of dust mite allergen Der p II and Der fII came in mensuration Air Conditioning filter screen dust and the suction cleaner collection dust.Say by Der p II and Der fII in PBS buffer extraction Air Conditioning filter screen dust and the suction cleaner collection dust as test sample with having.Adopt Der p II and Der fII monoclonal antibody parcel magnetic nanoparticle respectively, combine with testing sample, how anti-the Der p II, the Der fII that add alkali phosphatase enzyme mark again be, forms solid-phase coating antibody-antigen-enzyme labeling two anti-compounds behind 37 ℃ of incubations.Washing, separation back add substrate A MPPD as the chemiluminescence agent, and AMPPD is under the catalytic action of alkaline phosphatase, and mediator AMPD is produced in dephosphorization acid rapidly.The very fast decomposition of AMPD from the high energy excited state, is got back to low-energy stable state, ballistic phonon simultaneously, and this luminous stable, duration was 1 hour.Write down luminous intensity by the light quantum reading system, and calculate the content of allergen Der p II to be measured and Der fII from typical curve.Calculate dust mite allergen Der p II and Der fII total content in Air Conditioning filter screen dust and the suction cleaner collection dust respectively by the dust total amount.
The present invention not only provides the technological means of the detection of house dust mite pollution situation, and provides technical foundation for healthy household electrical appliances comprise the development of air-conditioning and suction cleaner.
Summary of the invention
The present invention is by magnetic nanoparticle enzyme-linked immunoassay technology, and the content of dust mite allergen Der p II and Der fII comes the house dust mite pollution situation is monitored in mensuration Air Conditioning filter screen dust and the suction cleaner collection dust.Say by Der p II and Der fII in PBS buffer extraction Air Conditioning filter screen dust and the suction cleaner collection dust as test sample with having.Adopt Der p II and Der fII monoclonal antibody parcel magnetic nanoparticle respectively, combine with testing sample, how anti-the Der p II, the Der fII that add alkali phosphatase enzyme mark again be, forms solid-phase coating antibody-antigen-enzyme labeling two anti-compounds behind 37 ℃ of incubations.Washing, separation back add substrate A MPPD as the chemiluminescence agent, and AMPPD is under the catalytic action of alkaline phosphatase, and mediator AMPD is produced in dephosphorization acid rapidly.The very fast decomposition of AMPD from the high energy excited state, is got back to low-energy stable state, ballistic phonon simultaneously, and this luminous stable, duration was 1 hour.Write down luminous intensity by the light quantum reading system, and calculate the content of allergen Der p II to be measured and Der fII from typical curve.Calculate dust mite allergen Der p II and Der fII total content in Air Conditioning filter screen dust and the suction cleaner collection dust respectively by the dust total amount.
The present invention not only provides the technological means of house dust mite pollution situation detection, and provides technical foundation for healthy household electrical appliances comprise the development of air-conditioning and suction cleaner.
Embodiment
Content of the present invention specifies by following embodiment:
Embodiment 1: the extraction of the collection of Air Conditioning filter screen dust and dust mite allergen Der p II and Der fII
Open the indoor air intake opening lid of family expenses split-type air conditioner, take out air filter screen gently, place it on the big clean blank sheet of paper, with clean brush down, collect and the weighing general assembly (TW) the brush of the dust on the filter screen.The dust that to always collect earlier grinds, add 200ulPBS (pH7.4) damping fluid by every milligram of dust again, stirred 30 minutes, it is changed in the collection tube, swung 24 hours by per minute 200 rotational oscillations 8 ℃ of conditions, pressed 5000g centrifugal 20 minutes, and collected centrifugal supernatant, remove floating thing with 0.45 μ m membrane filtration.Filter back liquid and be testing sample.
Embodiment 2: the extraction of dust mite allergen Der p II and Der fII in the suction cleaner collection dust
With the domestic vacuum cleaners of band HEPA filter screen to flooring, metope, bed down, places such as bed quilt, furniture surface carry out dust suction, take out dust collecting installation gently and dust wherein collected its general assembly (TW) of weighing together.The dust that to always collect earlier grinds, add 200ul PBS (pH7.4) damping fluid by every milligram of dust again, stirred 30 minutes, it is changed in the collection tube, swung 24 hours by per minute 200 rotational oscillations 8 ℃ of conditions, pressed 5000g centrifugal 20 minutes, and collected centrifugal supernatant, remove floating thing with 0.45 μ m membrane filtration.Filter back liquid and be testing sample.
Embodiment 3 Air Conditioning filter screen dusts extract Der p II and Der fII Determination on content in the sample
The testing sample 100ul that gets embodiment 1 extraction measures.Adopt Der p II, Der fII monoclonal antibody parcel magnetic nanoparticle respectively, combine with testing sample, how anti-the Der p II, the Der fII that add alkali phosphatase enzyme mark again be, forms solid-phase coating antibody-antigen-enzyme labeling two anti-compounds behind 37 ℃ of incubations.Washing, separation back add substrate A MPPD as the chemiluminescence agent, and AMPPD is under the catalytic action of alkaline phosphatase, and mediator AMPD is produced in dephosphorization acid rapidly.The very fast decomposition of AMPD from the high energy excited state, is got back to low-energy stable state, ballistic phonon simultaneously, and this luminous stable, duration was 1 hour.Write down luminous intensity by the light quantum reading system, and calculate the content of allergen Der p II to be measured and Der fII from typical curve.Typical curve is drawn and is adopted the Liu Zhigang professor leader's of life science institute of Shenzhen University seminar to make standard items by the Der p II and the Der fII recombinant protein of gene engineering expression.Calculate dust mite allergen Der p II and Der fII total content in the Air Conditioning filter screen dust respectively by the dust total amount.
Embodiment 4 suction cleaner are collected dust and are extracted Der p II and Der fII Determination on content in the sample
The testing sample 100ul that gets embodiment 2 extractions measures.Adopt Der p II, Der fII monoclonal antibody parcel magnetic nanoparticle respectively, combine with testing sample, how anti-the Der p II, the Der fII that add alkali phosphatase enzyme mark again be, forms solid-phase coating antibody-antigen-enzyme labeling two anti-compounds behind 37 ℃ of incubations.Washing, separation back add substrate A MPPD as the chemiluminescence agent, and AMPPD is under the catalytic action of alkaline phosphatase, and mediator AMPD is produced in dephosphorization acid rapidly.The very fast decomposition of AMPD from the high energy excited state, is got back to low-energy stable state, ballistic phonon simultaneously, and this luminous stable, duration was 1 hour.Write down luminous intensity by the light quantum reading system, and calculate the content of allergen Der p II to be measured and Der fII from typical curve.Typical curve is drawn and is adopted the Liu Zhigang professor leader's of life science institute of Shenzhen University seminar to make standard items by the Der p II and the Der fII recombinant protein of gene engineering expression.Calculate suction cleaner respectively by the dust total amount and collect dust mite allergen Der p II and Der fII total content in the dust.
Claims (5)
1, a kind of method that detects dust mite allergen in Air Conditioning filter screen dust and the suction cleaner collection dust.The dust that specifically passes through the collection of PBS buffer extraction is as test sample.Adopt Der p II, Der f II monoclonal antibody parcel magnetic nanoparticle respectively, combine with testing sample, how anti-the Der p II, the Der f II that add alkali phosphatase enzyme mark again be, forms solid-phase coating antibody-antigen-enzyme labeling two anti-compounds behind 37 ℃ of incubations.Washing, separation back add substrate A MPPD as the chemiluminescence agent, and AMPPD is under the catalytic action of alkaline phosphatase, and mediator AMPD is produced in dephosphorization acid rapidly.The very fast decomposition of AMPD from the high energy excited state, is got back to low-energy stable state, ballistic phonon simultaneously, and this luminous stable, duration was 1 hour.By light quantum reading system record luminous intensity, and calculate the content of allergen Der p II and Der f II the testing sample from typical curve.
2, the method for dust mite allergen Der p II and Der f II content in the method detection Air Conditioning filter screen dust according to claim 1.
3, the method for dust mite allergen Derp II and Der f II content in the detection suction cleaner collection dust according to claim 1.
4, be applied to assess the test that air-conditioning is prevented and treated dirt mite effect according to claim 1 and 2 described methods.
5, want 1 and 3 described methods to be applied to assess the test that suction cleaner is removed the house dust mite effect according to right.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410051395 CN1614420A (en) | 2004-09-09 | 2004-09-09 | Method for inspecting dermatophgoides allergen on air conditioner screen and in dust in dust vacuum cleaner |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410051395 CN1614420A (en) | 2004-09-09 | 2004-09-09 | Method for inspecting dermatophgoides allergen on air conditioner screen and in dust in dust vacuum cleaner |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1965748B (en) * | 2005-11-02 | 2010-05-26 | 松下电器产业株式会社 | Electric vacuum cleaner |
| CN102331492A (en) * | 2011-06-14 | 2012-01-25 | 浙江大学 | Method for detecting mite allergen specific antibody in blood serum |
| CN113970136A (en) * | 2021-11-22 | 2022-01-25 | 张云飞 | Indoor purifier with disinfect and remove flavor function |
-
2004
- 2004-09-09 CN CN 200410051395 patent/CN1614420A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1965748B (en) * | 2005-11-02 | 2010-05-26 | 松下电器产业株式会社 | Electric vacuum cleaner |
| CN102331492A (en) * | 2011-06-14 | 2012-01-25 | 浙江大学 | Method for detecting mite allergen specific antibody in blood serum |
| CN113970136A (en) * | 2021-11-22 | 2022-01-25 | 张云飞 | Indoor purifier with disinfect and remove flavor function |
| CN113970136B (en) * | 2021-11-22 | 2023-09-19 | 惠特科学技术有限公司 | Indoor purifier with sterilization and deodorization functions |
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