CN102095856A - Superoxide dismutase (SOD) rapid detection card and detection method thereof - Google Patents
Superoxide dismutase (SOD) rapid detection card and detection method thereof Download PDFInfo
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- CN102095856A CN102095856A CN2010106014885A CN201010601488A CN102095856A CN 102095856 A CN102095856 A CN 102095856A CN 2010106014885 A CN2010106014885 A CN 2010106014885A CN 201010601488 A CN201010601488 A CN 201010601488A CN 102095856 A CN102095856 A CN 102095856A
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Abstract
The invention relates to a superoxide dismutase (SOD) rapid detection card and a detection method thereof and belongs to the technical field of enzyme detection. A test strip is arranged in a shell of the SOD rapid detection card and is formed by sticking a sample pad, a colloidal gold membrane, a nitrocellulose membrane and a water absorbent membrane sequentially on a supporting rear board; the colloidal gold membrane is a glass fiber membrane which comprises anti-SOD antibody colloidal gold markers; and the nitrocellulose membrane is provided with two display marking belts, namely one display marking belt is a detection belt which comprises the SOD, and the other display marking belt is a quality control belt which comprises an anti-rabbit antibody or an anti-rat antibody. The SOD rapid detection card has the advantages of directly detecting the content of the SOD in specimens of SOD food, SOD health products and SOD cosmetics by an immunological method, is easy to prepare, can be conveniently and fast used and has accurate results.
Description
Technical field
The present invention relates to a kind of superoxide dismutase rapid detection card and detection method thereof, is a kind of appliance that detects superoxide dismutase, belongs to the enzymatic detection techniques field.
Background technology
Hepatocuprein (Superoxide dismutase is called for short SOD) extensively is present in the metal fermentoid that biosome includes Cu, Zn, Mn, Fe.It can remove unnecessary superoxide anion in the body as free radical scavenger important in the biosome, is playing an important role aspect the defence biosome oxidative damage.Ion (O
2 --) be human body oxygen metabolism product, it in vivo excess accumulation can cause inflammation, diseases such as tumour, color spot precipitation, aging, the generation of numerous disease and form relevant in superoxide anion and the biosome.
The vigour-testing method of SOD is a lot, and common have chemical method, immunization and an isoelectric focusing method.Wherein the chemical method application is the most general, and chemical ratio juris mainly is to utilize some compound can produce coloured intermediate and O in the autoxidation process
2 --, utilize SOD to decompose and calculate enzyme activity indirectly.In chemical method, the most frequently used have xanthine oxidase, pyrogallol method, chemoluminescence method, adrenaline method, NBT-reducing process, photochemistry TRAP, a Cyte reducing process etc.
Although it is a lot of to measure the enzyme of SOD method alive, but because the different and concrete condition determination of assay method is different, the enzyme work that same sample is measured in the different experiments chamber differs greatly, have in addition differ nearly thousand times, even same people adopts distinct methods to measure same sample gained data to differ also bigger.
For this reason, the present invention sends out bright a kind of immunological detection method based on the superoxide dismutase proteantigen, take immunology competition ratio juris, a kind of quick detection reagent of association colloid gold labelling technique and immunochromatographic method design, can be fast the superoxide dismutase in the test sample specifically.Superoxide dismutase content in the sample is judged in the colour developing that has or not according to test card sample detection line (T) and nature controlling line (C).Reaction is to utilize the superoxide dismutase antigenicity and a kind of method of protein detection of setting up, has the advantage of high specificity, the flase drop problem that can avoid the false positive of other detection method to cause.Simultaneously test card is simple to operate need not special instrument and equipment fast, both can carry out in the laboratory, also can be in the supermarket, family, workshop etc. measure on the spot.Because sample size needs less, easy fast, be applicable to the examination of great amount of samples.
Summary of the invention
The object of the present invention is to provide a kind of appliance that can detect superoxide dismutase, directly the method for applied immunology directly detects the content of food, health products and cosmetic sample superoxide dismutase, overcome the defective of existing superoxide dismutase activity detection method, set up based on the specific detection method of immunology.
Technical scheme of the present invention: a kind of superoxide dismutase rapid detection card, in the test card shell 1 of rectangular flat shell shape, test-strips 2 is set, the test card case surface has the fenestra 3 of detection and well 4; Test-strips 2 is formed by pasting sample pad 8, collaurum film 7, nitrocellulose filter 6 and absorbing membrane 9 thereon successively by adhesive sticker on the supporting backboard 5; The stacked stickup nitrocellulose filter 6 in supporting backboard middle part, one endlap of supporting backboard is put and is pasted absorbing membrane 9, the stacked stickup sample pad 8 of the other end, absorbing membrane is inner to overlap with nitrocellulose filter respectively separately with sample pad the inner, at the clinch of sample pad and nitrocellulose filter, insert and put between the two and paste one section collaurum film 7; The collaurum film is the glass fibre membrane that contains anti-superoxide dismutase antibody colloidal gold mark, there is one to detect band and a quality control band on the nitrocellulose filter, be successively: the detection that contains superoxide dismutase is with 10, the quality control band 11 that contains anti-rabbit antibody or anti-mouse antibody, test-strips is inserted in the test card shell, sample pad is over against well, and nitrocellulose filter is over against detecting fenestra.
The content of superoxide dismutase in the direct test sample of the method for applied immunology, test sample are superoxide dismutase SOD food, SOD health products and SOD cosmetics, for various test sample with different disposal routes:
Take by weighing 1.00g superoxide dismutase SOD food, SOD health products or SOD cosmetics solid sample and place glass mortar, add 9.0mL distilled water and grind 5min; If the sample liquid clarification is then directly got stoste and is measured; If the sample liquid muddiness then moves into the 10mL centrifuge tube, with a small amount of distilled water flushing mortar, washing is incorporated in the centrifuge tube, and adding distil water, is got supernatant and measured through the centrifugal 15min of 4000r/min to scale.
When detection be with 10 and quality control band 11 manifest brownish red trace band simultaneously, testing result is negative, represents that superoxide dismutase content does not exceed standard in the detected sample;
Be with 10 Show Colors not when detection, have only quality control band 11 to manifest the brownish red trace, testing result is positive, represents superoxide dismutase content overproof in the detected sample;
When detection be with 10 and quality control band 11 all do not develop the color, show that then test-strips lost efficacy.
Advantage of the present invention is to detect the superoxide dismutase that contains in the samples such as SOD food, SOD health products, SOD cosmetics, is easy to preparation and saves testing cost, and is easy to use, detects fast, and highly sensitive, the result is accurate.
Description of drawings
Fig. 1 superoxide dismutase rapid detection card shell and test-strips arrangement figure in the enclosure.1, shell, 2, test-strips, 3, detect fenestra, 4, well.
Fig. 2 detects and shows the trace synoptic diagram on the nitrocellulose filter of strip.6, nitrocellulose filter, 10, detect band, 11, quality control band.
The detection strip structural representation of Fig. 3 superoxide dismutase rapid detection card.5, supporting backboard, 6, nitrocellulose filter, 7, the collaurum film, 8, sample pad, 9, absorbing membrane.
Embodiment
The present invention further specifies as follows in conjunction with specific embodiments referring to accompanying drawing:
Embodiment 1
A kind of superoxide dismutase rapid detection card, it is that test-strips 2 is set in test card shell 1, the setting of the structure of test card shell 1 and test-strips 2 is referring to accompanying drawing 1, test card shell 1 is rectangular flat shell shape shell, long 70mm, wide 20mm, thick 5mm, shell wall thickness 1mm is made by engineering plastics.Test card shell 1 is connected with the shell block two halves by cap and constitutes, and the fenestra 3 of detection and well 4 are arranged on test card shell 1 cap.Test-strips 2 is placed in the shell block of test card shell 1.
Test-strips 2 is fillet thin slices of sandwich construction, and sheet is about 60mm, the wide about 4mm of sheet, the about 2.5mm of thickness.Test-strips 2 is sandwich constructions: bottom is a supporting backboard 5, is the igelite thin slice, thick about 0.5mm, long 60mm, wide 4mm; The stacked stickup nitrocellulose filter 6 in supporting backboard 5 middle parts, cellulose nitrate thickness 0.5mm, long 20-25mm, wide 4mm.Two isolated horizontal demonstration trace bands are arranged on the nitrocellulose filter 6, show in the trace band referring to 2, two in accompanying drawing, one is to detect to be with 10, contains superoxide dismutase; Another is a quality control band 11, contains anti-rabbit antibody or anti-mouse antibody.Stacked stickup absorbing membrane 9 on supporting backboard 5 one ends, stacked stickup sample pad 8 on supporting backboard 5 the other end, absorbing membrane 9 the inners overlap with nitrocellulose filter 6 one ends respectively separately with sample pad 8 the inners, at the clinch of sample pad 8 with nitrocellulose filter 6, insert and put between the two and paste one section collaurum film 7, collaurum film 7 is the glass fibre membranes that contain anti-superoxide dismutase monoclonal antibody or polyclonal antibody colloid gold label thing.Collaurum film 7 thick 0.5-0.8mm, long 5mm, wide 4mm.Sample pad 8 thick 0.5-0.8mm, long 20mm, wide 4mm.Absorbing membrane 9 thick 0.5-0.8mm, long 16-20mm, wide 4mm.Collaurum film 7, sample pad 8 and absorbing membrane 9 are 1-2mm with the nitrocellulose filter lap of splice.Absorbing membrane 9 materials are thieving paper, and sample pad 8 materials are water adsorption glass tunica fibrosas.
Superoxide dismutase test card of the present invention is used for detecting the detection of the superoxide dismutase of samples such as SOD food, SOD health products, SOD cosmetics.
The disposal route of sample is:
Test sample is superoxide dismutase SOD food, SOD health products and SOD cosmetics, for various test sample with different disposal routes:
Take by weighing 1.00g superoxide dismutase SOD food, SOD health products or SOD cosmetics solid sample and place glass mortar, add 9.0mL distilled water and grind 5min; If the sample liquid clarification is then directly got clarified solution and is measured; If the sample liquid muddiness then moves into the 10mL centrifuge tube, with a small amount of distilled water flushing mortar, washing is incorporated in the centrifuge tube, and adding distil water, is got supernatant and measured through the centrifugal 15min of 4000r/min to scale.
When using the superoxide dismutase rapid detection card to detect, earlier sample liquid to be detected is instilled into from well 4 on the sample pad 8 of superoxide dismutase rapid detection card 2, because syphonic effect principle, drive contained anti-superoxide dismutase monoclonal antibody of sample liquid to be detected and collaurum film 7 or polyclonal antibody colloid gold label things together to nitrocellulose filter 6 diffusions, observations in 5-10 minute.
The key reaction of test card is immunologic antigen and antibody response, the antibody of the colloid gold label that on nitrocellulose filter, moves, on calibration tape respectively with contain superoxide dismutase, and the anti-rabbit antibody of quality control band or anti-mouse antibody reaction forms the brownish red band.If have superoxide dismutase to be checked to exist in the sample, after sample adds, promptly react, and can not react with contained superoxide dismutase, thereby do not develop the color with detection with antibody.The result of key reaction has following 3 kinds of situations:
1, when show to detect be with 10 and quality control band 11 manifest brownish red trace band simultaneously, illustrate that superoxide dismutase content does not exceed standard in the detected sample.
2, be with 10 Show Colors not when detection, have only quality control band 11 to manifest the brownish red trace, testing result is positive, and superoxide dismutase content overproof in the detected sample is described.
3, when detection be with 10 and quality control band 12 all do not develop the color, show that then detecting strip lost efficacy.
Claims (2)
1. superoxide dismutase rapid detection card, it is characterized in that: in the test card shell (1) of rectangular flat shell shape test-strips (2) is set, the test card case surface has detection fenestra (3) and well (4); Test-strips (2) is upward pasted sample pad (8), collaurum film (7), nitrocellulose filter (6) and absorbing membrane (9) thereon successively by adhesive sticker by supporting backboard (5) and is formed; Supporting backboard middle part stacked stickup nitrocellulose filter (6), one endlap of supporting backboard is put and is pasted absorbing membrane (9), the stacked stickup sample pad of the other end (8), absorbing membrane is inner to overlap with nitrocellulose filter respectively separately with sample pad the inner, at the clinch of sample pad and nitrocellulose filter, insert and put between the two and paste one section collaurum film (7); The collaurum film is the glass fibre membrane that contains anti-superoxide dismutase antibody colloidal gold mark, there is one to detect band and a quality control band on the nitrocellulose filter, be successively: the detection band (10) that contains superoxide dismutase, the quality control band (11) that contains anti-rabbit antibody or anti-mouse antibody, test-strips is inserted in the test card shell, sample pad is over against well, and nitrocellulose filter is over against detecting fenestra.
2. the detection method of the described superoxide dismutase rapid detection card of claim 1, the content that it is characterized in that superoxide dismutase in the direct test sample of method of applied immunology, test sample is superoxide dismutase SOD food, SOD health products and SOD cosmetics, for various test sample with different disposal routes:
Take by weighing 1.00g superoxide dismutase SOD food, SOD health products or SOD cosmetics solid sample and place glass mortar, add 9.0mL distilled water and grind 5min; If the sample liquid clarification is then directly got clarified solution and is measured; If the sample liquid muddiness then moves into the 10mL centrifuge tube, with a small amount of distilled water flushing mortar, washing is incorporated in the centrifuge tube, and adding distil water, is got supernatant and measured through the centrifugal 15min of 4000r/min to scale;
When detection be with 10 and quality control band 11 manifest brownish red trace band simultaneously, testing result is negative, represents that superoxide dismutase content does not exceed standard in the detected sample;
Be with 10 Show Colors not when detection, have only quality control band 11 to manifest the brownish red trace, testing result is positive, represents superoxide dismutase content overproof in the detected sample;
When detection be with 10 and quality control band 11 all do not develop the color, show that then test-strips lost efficacy.
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| CN2010106014885A CN102095856A (en) | 2010-12-23 | 2010-12-23 | Superoxide dismutase (SOD) rapid detection card and detection method thereof |
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| CN2010106014885A CN102095856A (en) | 2010-12-23 | 2010-12-23 | Superoxide dismutase (SOD) rapid detection card and detection method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103336132A (en) * | 2013-07-08 | 2013-10-02 | 无锡安迪生物工程有限公司 | Detection method of lactoferrin in tears and dedicated colloidal gold detecting card thereof |
| CN106957891A (en) * | 2017-05-16 | 2017-07-18 | 重庆师范大学 | The purposes and kit of gold nanoclusters and copper-zinc superoxide dismutase |
| CN109374885A (en) * | 2018-10-22 | 2019-02-22 | 中国农业科学院作物科学研究所 | A kind of detection method of colloid gold test paper, preparation method and Rice Seed Vigor |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101625368A (en) * | 2009-08-04 | 2010-01-13 | 无锡安迪生物工程有限公司 | Casein rapid detection card and detection method thereof |
-
2010
- 2010-12-23 CN CN2010106014885A patent/CN102095856A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101625368A (en) * | 2009-08-04 | 2010-01-13 | 无锡安迪生物工程有限公司 | Casein rapid detection card and detection method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 黄浦雁: "食品中SOD检测方法的研究", 《食品与发酵工业》, no. 3, 31 December 1996 (1996-12-31) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103336132A (en) * | 2013-07-08 | 2013-10-02 | 无锡安迪生物工程有限公司 | Detection method of lactoferrin in tears and dedicated colloidal gold detecting card thereof |
| CN106957891A (en) * | 2017-05-16 | 2017-07-18 | 重庆师范大学 | The purposes and kit of gold nanoclusters and copper-zinc superoxide dismutase |
| CN109374885A (en) * | 2018-10-22 | 2019-02-22 | 中国农业科学院作物科学研究所 | A kind of detection method of colloid gold test paper, preparation method and Rice Seed Vigor |
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Application publication date: 20110615 |