CN1517709A - Novel competition method for detecting antigen by immune chromatographic reaction on microporous membrane medium - Google Patents
Novel competition method for detecting antigen by immune chromatographic reaction on microporous membrane medium Download PDFInfo
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- CN1517709A CN1517709A CNA031148883A CN03114888A CN1517709A CN 1517709 A CN1517709 A CN 1517709A CN A031148883 A CNA031148883 A CN A031148883A CN 03114888 A CN03114888 A CN 03114888A CN 1517709 A CN1517709 A CN 1517709A
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- antigen
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Abstract
A novel competition method for detecting the content of antigen by immunochromatographic reaction on micro pore membrane medium features that the number of labelling antibodies is far greater than that of the antibodies needed by binding all antigen in specimen, the number of the coated antigens is far greater than that needed by binding all free labelling antibodies, and the optical signal at the detection position can directly reflect the content of antigen. Its advantage is high sensitivity and correctness.
Description
In the immune diagnostic method,, can make sample solution from one end to the other side move chromatography and carry out immune response, set up fast simple detection method by the microporous barrier medium.The very early pregnancy fast detecting strip that utilizes the colloid gold particle labelling technique to produce at present is exactly that this method is produced.The fast detecting of a lot of drugs has also been set up a kind of method of competing the immunochromatography reaction.Colloidal gold-labeled method is a kind of method wherein.In the collaurum detection technique, the method that comparative maturity is used has three kinds at present: the immune response of sandwich method chromatography detects antigen; The immune response of indirect method chromatography detects antibody; The immune response of conventional competition law chromatography detects antigen.The main points that above-mentioned conventional competition law immunity reflection detects the antigen method are to set the antigenic content that the minimum requirements in the sample detects earlier, make the labelled antibody amount of bag quilt be less than needs in conjunction with the intact needed antibody amount of this content antigen then.If requirement value of detecting of antigen is very low, the labelled antibody amount also just must seldom just be difficult in the detection position and produce necessary color.So conventional competition law immunochromatography reaction is difficult to detect the antigen of trace in the sample
The optical signalling detection antigen method of the application's novel competition law microporous barrier medium immunochromatography reaction has overcome the shortcoming in the above method, can detect the antigen of utmost point low content.Now illustrate as follows below the combination:
Position A to a section of position E be micropore shape film medium, micropore shape film can be a cellulose nitrate, cellulose acetate, nylon, glass fibre, filter paper, and the film that is full of micropore made of material such as other high molecular polymers, this medium can be combined by single kind or multiple film, and sample can directly be applied on the micropore shape film medium.
Position A is the sample point of release, and sample can be all kinds of solution that may contain detected antigen, as urine, blood, serum, saliva etc.Antigen is all and the antibody that is labeled constitutes the material in secure bond site.
B place, position fixedly is coated with the excessive antibody that is labeled, i.e. the binding capacity that needs of the antigen that the quantity of antibody may maximum in the sample.The material that this antibody is labeled can be a colloid gold particle, latex particle, certain enzyme, any in chemiluminescence or the fluorescence molecule etc.
C place, position fixed packet is had the antigen in secure bond site by excessive and antibody, promptly the quantity of antigen far more than whole labelled antibodies by antigen in conjunction with the intact amount that needs.This antigen can be simple antigen, or combines with the big molecule of other biological but keep secure bond site with labelled antibody.
D place, position fixedly is coated with excessive energy incorporation of markings antibody and antigen bond biomacromolecule.
When discharging the sample solution that contains antigen at position A, solution is carrying antigen and is moving towards position B, and the excessive labelled antibody part at position B place is by the antigen combination, and a part of labelled antibody then is free in the solution, all continues towards position C travel motion.At position C place, free labelled antibody is moved no longer forward, and directly or indirectly shows optical signalling by the whole combinations of antigen of excessive fixed packet quilt.Position B place by sample in the antigen combination labelled antibody then because binding site saturated, then continue to cross position C, move towards position D.By the big molecule trapping of other biological at position D place, directly or indirectly show optical signalling then.When not having antigen in the sample solution, the color signal at position C place is influenced hardly, and D place in position does not then have signal.Therefore the signal power at D place, position directly reflects the content of antigen.
The competition law immunochromatography detection for Morphine in Urine now is described as follows for embodiment: film medium is by German S﹠amp; S company 33 type glass fibre membranes, German S﹠amp; The AE99 of S company type nitrocellulose filter, German S﹠amp; S company 470 class absorbent filter is formed.Be coated with the morphine antibody of colloid gold particle mark on the 33 type glass fibre membranes.Fixedly be coated with the crosslinked bond of morphine and BSA on the AE99 nitrocellulose filter, be coated with sheep anti-mouse igg thereafter again and resist more, the full wafer nitrocellulose filter seals with BSA solution and handles and drying.In room temperature is 25 degree, and relative humidity is connecting above three kinds of materials successively and to stick on U.S. G﹠amp in 30% clean environment; On the biological diagnosis type PET of the L company gum base plate.
Directly be released on the 33 type glass fibre membranes with urine sample, the big molecule of morphine and other biological is travel motion forward, arrive on the nitrocellulose filter after, morphine is formed bond with the part colloid gold label antibody that wherein wraps quilt.Bond continues travel motion forward with other free colloid gold label antibody, and when being coated with the position of morphine and the crosslinked bond of BSA, whole combined the fixing of all free colloid gold label antibody show danger signal.The bond of morphine and colloid gold label antibody then continues to move forward, and the sheep anti-mouse igg of the bag quilt that is fixed resists more is caught, and manifests danger signal.This danger signal directly is directly proportional with morphine content in the sample.The big molecule of other biological then continues travel motion, is absorbed by the 470 class absorbent filter at last.
Claims (8)
1. novel competition law microporous barrier medium immunochromatography reaction by optical signalling detection antigen, after it is characterized in that excessive labelled antibody combines antigen in the sample solution, free labelled antibody is analyzed antigen by the antigen capture of the excessive bag quilt of microporous barrier medium by the optical signalling that antigen in the sample solution and labelled antibody produce.
2. the sample in the claim 1 can be a urine, blood, saliva, all kinds of solution of serum.
3. labelled antibody can directly or indirectly be presented the material mark of optical signalling by all in the detection position in the claim 1, as colloid gold particle, and latex particle, horseradish peroxidase, chemiluminescence or fluorescence molecule.
4. the film in the microporous barrier medium can be a cellulose nitrate in the claim 1, cellulose acetate, nylon, glass fibre, filter paper, and the material that other high molecular polymers etc. are full of micropore makes, and may be through handling obtaining particular characteristic, and the microporous barrier medium can be planted or multiple above microporous barrier be combined by single.
Excessive labelled antibody in the claim 1 be may high-load with respect in the sample the combined intact amount of antigen.
The antigen of the excessive bag quilt in the claim 1 be relatively with the amount of the combined intact needs of all labelled antibodies, this antigen can be simple antigen, or with the macromolecular bond of other biological, but keep and the immune binding site of labelled antibody.
7. the optical signalling in the claim 1 can also can be measured with other optical detecting instruments by the eyes Direct observation.
8. claimed other biological analysis and detection device or instrument of claim according to this method design.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031148883A CN1517709A (en) | 2003-01-14 | 2003-01-14 | Novel competition method for detecting antigen by immune chromatographic reaction on microporous membrane medium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031148883A CN1517709A (en) | 2003-01-14 | 2003-01-14 | Novel competition method for detecting antigen by immune chromatographic reaction on microporous membrane medium |
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| Publication Number | Publication Date |
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| CN1517709A true CN1517709A (en) | 2004-08-04 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNA031148883A Pending CN1517709A (en) | 2003-01-14 | 2003-01-14 | Novel competition method for detecting antigen by immune chromatographic reaction on microporous membrane medium |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102393464A (en) * | 2011-10-25 | 2012-03-28 | 广东省药品检验所 | Preparation method of test strip for morphine detection and application thereof |
| US8956879B2 (en) | 2007-10-04 | 2015-02-17 | Panasonic Healthcare Co., Ltd. | Analysis device and method using the same |
| CN105324668A (en) * | 2013-08-12 | 2016-02-10 | 古河电气工业株式会社 | Silica particles, each of which has reactive functional group on surface, and method for producing same |
| CN109444406A (en) * | 2018-11-09 | 2019-03-08 | 深圳市众循精准医学研究院 | Test strips of quantitative detection marker cyfra21-1 and preparation method thereof and detection method |
| CN110244065A (en) * | 2019-05-16 | 2019-09-17 | 北京化工大学 | The Microfluidic Immunoassay Chip and its preparation method and application of 1 albumen of progesterone receptor membrane component in a kind of detection blood |
-
2003
- 2003-01-14 CN CNA031148883A patent/CN1517709A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8956879B2 (en) | 2007-10-04 | 2015-02-17 | Panasonic Healthcare Co., Ltd. | Analysis device and method using the same |
| CN103424543B (en) * | 2007-10-04 | 2015-04-29 | 松下健康医疗器械株式会社 | Analysis method using analysis device |
| CN102393464A (en) * | 2011-10-25 | 2012-03-28 | 广东省药品检验所 | Preparation method of test strip for morphine detection and application thereof |
| CN102393464B (en) * | 2011-10-25 | 2014-01-15 | 广东省药品检验所 | Preparation method of test strip for morphine detection and application thereof |
| CN105324668A (en) * | 2013-08-12 | 2016-02-10 | 古河电气工业株式会社 | Silica particles, each of which has reactive functional group on surface, and method for producing same |
| CN109444406A (en) * | 2018-11-09 | 2019-03-08 | 深圳市众循精准医学研究院 | Test strips of quantitative detection marker cyfra21-1 and preparation method thereof and detection method |
| CN110244065A (en) * | 2019-05-16 | 2019-09-17 | 北京化工大学 | The Microfluidic Immunoassay Chip and its preparation method and application of 1 albumen of progesterone receptor membrane component in a kind of detection blood |
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