CN203870107U - Group A streptococcus quantum dot immunochromatography test reagent card - Google Patents
Group A streptococcus quantum dot immunochromatography test reagent card Download PDFInfo
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- CN203870107U CN203870107U CN201420217173.4U CN201420217173U CN203870107U CN 203870107 U CN203870107 U CN 203870107U CN 201420217173 U CN201420217173 U CN 201420217173U CN 203870107 U CN203870107 U CN 203870107U
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The utility model discloses a group A streptococcus quantum dot immunochromatography test reagent card, and aims to provide a group A streptococcus quantum dot immunochromatography test reagent card which is simple and quick in detecting method, high in sensitivity and reliable in detection result. The test reagent card comprises a box body (6) with a box cover, wherein a piece of test paper is arranged in the box body (6); a sample adding hole (71) and a result watch window (72) are formed in the box cover (61) of the box body (6); the test paper comprises a baseplate (1); a sample cushion (2), a group A streptococcus resistant antibody quantum dot marking cushion (3), a coating film (4) and a water-absorbing cushion (5) are connected onto the baseplate (1) in sequence; a group A streptococcus antibody test area T line and a goat anti-rabbit polyclonal antibody quality control area C line are arranged on the coating film (4), and are arranged in parallel. The group A streptococcus quantum dot immunochromatography test reagent card belongs to the field of fluorescence immunodetection.
Description
Technical field
The utility model discloses a kind of detection reagent card, specifically, is the streptococcic detection reagent card of A family in the pharyngeal swab of a kind of mensuration, the invention also discloses the preparation method of this detection reagent card, belongs to fluorescence immunoassay detection field.
Background technology
One of common disease substance of children's upper respiratory tract infection, in children with respiratory infection, A family streptococcus positive rate is about 23.2%.As accepted not in time anti-infective therapy, comprise the allergic disease causing after acpuei pharyngitis, purulence born of the same parents disease, TSS, aggressive fascitis, scarlet fever, pneumonia, erysipelas and the streptococcal infection of A family, as acute rheumatic fever, rheumatic heart disease, acute glomerulonephritis.General A family streptococcus authentication method mainly contains clinical experience examination, bacterial cultivation at present.
On the low side according to the streptococcic accuracy rate of clinical experience diagnosis A family, according to statistics, even if the accuracy rate of veteran diagnosis also only has 45%-75%.Bacterial cultivation is A family streptococcus diagnosis " goldstandard ", and the method is that the throat swab of taking is inoculated in and on blood agar plate, hatches 24～48h, 10%CO
2or under anaerobic condition, can increase the streptococcic separation rate of A family, finally take serological method to differentiate A family streptococcus and other β hemolytic streptococcus, particularly C Zu He G family, but the time needing is longer.
Utility model content
For above-mentioned deficiency, it is a kind of simple to operate that the purpose of this utility model is to provide, and rapidly, the reliable A of testing result family streptococcus quantum dot immune chromatography detects reagent card in diagnosis.
For solving the problems of the technologies described above, the technical scheme that the utility model provides is such: this A family streptococcus quantum dot immune chromatography detects reagent card, comprise the box body with lid, be provided with Test paper in box body, the lid of described box body is provided with well and result watch window; Described Test paper comprises base plate, on base plate, be connected and have sample pad, the quantum dot-labeled pad of anti-A family's streptococcus antibody, coated film and adsorptive pads successively, described coated film is provided with Yi Tiao A family streptococcus antibody test district T line and a goat-anti rabbit polyclonal antibody Quality Control district C line, and two line parallels are arranged.
Above-mentioned A family streptococcus quantum dot immune chromatography detects reagent card, described coated film two ends respectively with adsorptive pads He Kang A family's streptococcus antibody mutual overlapping connection of quantum dot-labeled pad, on the quantum dot-labeled pad of Kang A family streptococcus antibody, overlayed sample pad.
Further, above-mentioned A family streptococcus quantum dot immune chromatography detects reagent card, and described sample pad is glass fiber sample pad.
Further, above-mentioned A family streptococcus quantum dot immune chromatography detects reagent card, and described coated film is nitrocellulose filter.
Further, above-mentioned A family streptococcus quantum dot immune chromatography detects reagent card, and the described quantum dot-labeled pad of anti-A family's streptococcus antibody is coated quantum dot-labeled A family streptococcus antibody polyester cellulose film.
Further, above-mentioned A family streptococcus quantum dot immune chromatography detects reagent card, sample pad is pressed in 1/3～1/4 place of the quantum dot-labeled pad of anti-A family's streptococcus antibody, the quantum dot-labeled pad of anti-A family's streptococcus antibody is pressed in the 1mm place of coated film, adsorptive pads is pressed in coated film 1～2.5mm place, the streptococcus antibody test district T of A family linear distance coated film left end 10mm, goat-anti rabbit polyclonal antibody Quality Control district C linear distance coated film right-hand member 10mm, C, T distance between centers of tracks 4～7mm.
Compared with prior art, A provided by the utility model family streptococcus quantum dot immune chromatography detects reagent card tool and has the following advantages:
The technical scheme that the utility model provides, utilize the specific binding reaction of two kinds of antibody, form special " sandwich " structure, i.e. " antibody-Ag-Ab " immune complex, binding immunoassay chromatographic technique, application fluorescence immune chromatography method detects A family streptococcal antigen, and matches with hand-held uviol lamp, increases substantially the streptococcic recall rate of A family.
This method is simple to operate, and diagnosis is rapid, and high specificity is highly sensitive, easily stores, and timing operating personnel that need not be special, coordinate simple instrument and equipment, can obtain a result.Be conducive to like this medical personnel and catch golden hour, avoid antibiotic abuse, reduce the allergic probability of A family streptococcus simultaneously.
Accompanying drawing explanation
Fig. 1 is the detection reagent card structural representation that the utility model provides;
Fig. 2 is the lid vertical view of the detection reagent card that provides of the utility model;
Fig. 3 is the Test paper structural representation that the utility model provides;
Fig. 4 is the Test paper vertical view that the utility model provides;
Fig. 5 is the test card result of determination that provides of the utility model schematic diagram when negative;
Fig. 6 is the test card result of determination that provides of the utility model schematic diagram when positive.
Wherein: base plate 1, sample pad 2, the quantum dot-labeled pad 3 of anti-A family's streptococcus antibody, coated film 4, adsorptive pads 5, the streptococcus antibody test district T of A family line, goat-anti rabbit polyclonal antibody Quality Control district C line, box body 6, lid 61, well 71, result watch window 72.
Embodiment
Mode below in conjunction with specific embodiment is described in further detail claim of the present utility model, but do not form any restriction of the present utility model, the modification of the limited number of time that anyone makes within the scope of the utility model claim, still within claim scope of the present utility model.
Embodiment 1
A kind of A family streptococcus quantum dot immune chromatography that the utility model provides detects reagent card, consult Fig. 1 to Fig. 4, described detection reagent card comprises the box body 6 with lid 61, and in order to facilitate application of sample and to observe testing result, described lid 61 is provided with well 71 and result watch window 72.
In box body 6, be provided with Test paper, described Test paper comprises base plate 1, described base plate 1 is PVC plate, on base plate 1, be connected and have sample pad 2, the quantum dot-labeled pad 3 of anti-A family's streptococcus antibody, coated film 4 and adsorptive pads 5 successively, on coated film 4, be provided with Yi Tiao A family streptococcus antibody test district T line and a goat-anti rabbit polyclonal antibody Quality Control district C line, two line parallels are arranged.In order to reach testing result fast and accurately, described sample pad 2 preferred glass fibers sample pad; The described preferred nitrocellulose filter of coated film 4; The quantum dot-labeled pad 3 of described anti-A family's streptococcus antibody is coated quantum dot-labeled A family streptococcus antibody polyester cellulose film.
In particular, described coated film 4 overlays in base plate 1 middle part, the two ends of coated film 4 respectively with 5 He Kang A family streptococcus antibody mutual overlapping connections of quantum dot-labeled pad 3 of adsorptive pads, on the quantum dot-labeled pad 3 of Kang A family streptococcus antibody, overlayed sample pad 2.
Sample pad 2 is pressed in 1/3～1/4 left and right of the quantum dot-labeled pad 3 of anti-A family's streptococcus antibody, the quantum dot-labeled pad 3 of anti-A family's streptococcus antibody is pressed in the 1mm place of coated film 4, the streptococcus antibody test district T of A family linear distance coated film 4 left end 10mm, goat-anti rabbit polyclonal antibody Quality Control district C linear distance coated film 4 right-hand member 10mm, C, T distance between centers of tracks 4～7mm (preferably 5mm), 1～2.5mm (preferably 2mm) that adsorptive pads is pressed in coated film 4 locates.
This A family streptococcus quantum dot immune chromatography detects the preparation method of reagent card:
1) preparation method of sample pad is: by 0.01M phosphate buffer (the 0.01M sodium hydrogen phosphate: 0.01M sodium dihydrogen phosphate=81:19 (V:V)), after (pH7.4) soaking completely, dry or be dried under the environment of relative humidity≤35% for 37 ℃ that contains 0.5～5wt%BSA, 0.5～5wt% polyvinylpyrrolidone, 0.5～5wt% polyglycol, 0.1～5wt% Tween-20, bent La Tong-100 of 1～5wt% for sample pad.
Damping fluid is preferably 0.5wt%BSA in 0.01M phosphate buffer, 0.5wt% polyvinylpyrrolidone, 0.5wt% polyglycol, 0.25wt% Tween-20, bent La Tong-100 of 1wt%.
2) preparation method of the quantum dot-labeled pad of anti-A family's streptococcus antibody is:
1. use borate buffer (the 2.5mM borax: 10mM boric acid=4.5:5.5 (V:V)) QD525 of water-soluble carboxyl based quantum dot 8 μ M is diluted to 1 μ M of 10mM pH8.4.
2. get the quantum dot of 1mL1 μ M, add 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide hydrochloric acid solution of 30 μ L10mg/mL, room temperature reaction 1～2 hour.With the super filter tube ultrafiltration of 30kDa, remove unreacted 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide hydrochloric acid, draw remaining liq in super filter tube, add the borate buffer of 10mM to revert to volume to 1mL.
③Jiang Kang A family streptococcus antibody adds in the super filter tube of 100kDa, with 10mM borate buffer adjustment antibody content, is 4mg/mL.
4. by the amount of 200 μ g/1mL, the antibody that 2. step is prepared joins step 1. in quantum dot solution, room temperature reaction 1～2 hour.
5. by the amount of 10 μ L/1mL, to step, add the glycocoll of 10mg/mL in 3., seal the not activated carboxyl site of coupling antibody, reaction 30～60min.
6. the quantum dot solution of coupling antibody is joined in the super filter tube of 30kDa, add pH9.0 concentration 0.05M Tris-HCl solution substitutional solution buffer system.Repeat 3～5 times, draw the liquid in super filter tube, add Tris-HCl to final volume be 100 μ L.
7. by step in 6. to be dissolved in 4 ℃ of 1800g of liquid centrifugal, get supernatant.
8. in step, 7. in the supernatant of gained, add 100 μ L antibody to preserve liquid, described body is preserved liquid for containing 1%BSA, 20% sucrose, 10% trehalose, the 0.05M Tris-HCl damping fluid of 0.1% Tween-20 and 0.1% polyvinylpyrrolidone.
8. the solution of above-mentioned gained is drawn to film instrument with metal spraying and be sprayed onto on polyester cellulose film, discharge rate is 1～5 μ L/cm (preferably 2 μ L/cm), puts into 37 ℃ of baking ovens, dry 2～5 hours.
3) preparation method of coated film is:
1. goat anti-rabbit antibody is diluted to 1mg/mL with 0.02M phosphate buffer, adds 3% methyl alcohol, be Quality Control district working fluid.
②Jiang Kang A family streptococcus antibody is diluted to 1.2mg/mL with 0.02M phosphate buffer, adds 3% methyl alcohol, is detection zone working fluid.
3. by 1., the 2. solution of gained, with metal spraying, draw film instrument and be coated with on nitrocellulose membrane with 1 μ L/cm.Detection zone and Quality Control Interval Distance are 0.4～0.7cm (preferably 0.5cm), are positioned at nitrocellulose filter centre position.
4. by step 3. nitrocellulose membrane put into 37 ℃ of baking ovens, dried overnight.
4) assembling:
1. the stickup of nitrocellulose filter: open gently the diaphragm of NC film location for paste on PVC plate, NC film is evenly advanced from left to right, it is firmly sticked on PVC plate.
2. the stickup of adsorptive pads: PVC plate is laid on work top; Open gently the diaphragm of absorption pad location for paste on PVC plate, absorption pad is adhered thereto, and even, slight tumbling-type advances, and to add strong adhesive power, and prevents that bubble, absorption pad cover 1mm on NC film.
3. the stickup of quantum dot-labeled pad: quantum dot-labeled pad is cut out as 5mm * 300mm.Open gently the diaphragm of the quantum dot-labeled pad of NC film upper limb location for paste, the quantum dot-labeled pad of collaurum is adhered thereto, the same absorption pad of method.The quantum dot-labeled pad of collaurum covers 1mm on NC film.
4. the stickup of sample pad: dried sample pad is cut out as after 18mm * 300mm, open gently thin plate diaphragm topmost, sample pad is adhered to the quantum dot-labeled pad of collaurum top, the same absorption pad of method.Sample pad covers 2mm on the quantum dot-labeled pad of collaurum.
5. the reinforcing of quantum dot-labeled pad: will be connected adhesive tape and cut out as 12mm * 300mm, and paste sample pad place, top is apart from test strips sample pad end 15mm.
6. slitting be installed: the PVC plate pasting is cut into the wide test strips of 3.0mm (2.5～4.5mm).Each test strips is packed in plastic clip, each reagent card is placed in to aluminum foil bag, and add 1g drying agent 1 bag, heat sealing.
This A family streptococcus quantum dot immune chromatography detects the concrete using method of reagent card:
After sampling, in extraction tube, add each 3～4 of lysate A (1M sodium nitrite) and lysate B (2M acetic acid), put into after cotton swab extruding several times, standing 3～5min.
During detection, consult Fig. 5 and Fig. 6, sample lysate is splashed in the well of this detection reagent to 3～4, because detecting sample, capillarity will to water sucting plate end, move along Test paper, A family streptococcus in sample is after lysate cracking, Ke Yukang A family streptococcus polyclonal antibody fluorescence labeling probe generation specific bond, then continuing mobile antibody on being coated on nitrocellulose filter is combined, thereby the streptococcic polyclonal antibody identification of Bei Kang A family, there is the specific bond of double-antibody sandwich, with hand-held ultraviolet light irradiation watch window, there is quantum dot fluorescence band in detection zone, thereby carry out the evaluation of respiratory tract infection pathogeny by the A family streptococcus detecting in sample.In testing process, the fluorescence probe that is stranded in detection zone when up liquid level will show corresponding fluorescence.Setting A family streptococcus boundary value is 10
3cFU/mL, when Quality Control district and detection zone have fluorescence to occur, in interpret sample, the streptococcic content of A family surpasses 10
3cFU/mL, positive; When only having Quality Control district to have fluorescence, detection zone occurs without fluorescence, interpret sample Zhong Buhan A family streptococcus or content are lower than 10
3cFU/mL, negative.
When moving to goat-anti rabbit polyclonal antibody Quality Control district, no matter in sample, have or not A family streptococcus, the goat anti-rabbit igg that fluorescence probe all can be coated on nitrocellulose filter is combined delay, makes Quality Control district show fluorescence.Therefore Quality Control district produces and represents that operation is wrong or testing result is invalid without fluorescence, needs revision test.
For better explanation the beneficial effects of the utility model, provide the detection reagent card and traditional microbe growth detection method that adopt the utility model to provide below, the result contrast experiment that clinical experience is screened.
Test example 1
Claims (6)
1.Yi Zhong A family streptococcus quantum dot immune chromatography detects reagent card, comprise the box body (6) with lid, box body is provided with Test paper in (6), it is characterized in that, the lid (61) of described box body (6) is provided with well (71) and result watch window (72); Described Test paper comprises base plate (1), on base plate (1), be connected and have sample pad (2), the anti-A quantum dot-labeled pad of family's streptococcus antibody (3), coated film (4) and adsorptive pads (5) successively, described coated film (4) is provided with Yi Tiao A family streptococcus antibody test district T line and a goat-anti rabbit polyclonal antibody Quality Control district C line, and two line parallels are arranged.
2. A according to claim 1 family streptococcus quantum dot immune chromatography detects reagent card, it is characterized in that, described coated film (4) two ends respectively with mutual overlapping connection of adsorptive pads (5) the He Kang A quantum dot-labeled pad of family's streptococcus antibody (3), on the quantum dot-labeled pad of Kang A family streptococcus antibody (3), overlayed sample pad (2).
3. A according to claim 1 family streptococcus quantum dot immune chromatography detects reagent card, it is characterized in that, described sample pad (2) is glass fiber sample pad.
4. A according to claim 1 family streptococcus quantum dot immune chromatography detects reagent card, it is characterized in that, described coated film (4) is nitrocellulose filter.
5. A according to claim 1 family streptococcus quantum dot immune chromatography detects reagent card, it is characterized in that, the described quantum dot-labeled pad of anti-A family's streptococcus antibody is coated quantum dot-labeled A family streptococcus antibody polyester cellulose film.
6. A according to claim 1 family streptococcus quantum dot immune chromatography detects reagent card, it is characterized in that, sample pad (2) is pressed in 1/3～1/4 place of the anti-A quantum dot-labeled pad of family's streptococcus antibody (3), the anti-A quantum dot-labeled pad of family's streptococcus antibody (3) is pressed in the 1mm place of coated film (4), adsorptive pads (5) is pressed in coated film (4) 1～2.5mm places, A family streptococcus antibody test district T linear distance coated film (4) left end 10mm, goat-anti rabbit polyclonal antibody Quality Control district C linear distance coated film (4) right-hand member 10mm, C, T distance between centers of tracks 4～7mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420217173.4U CN203870107U (en) | 2014-04-29 | 2014-04-29 | Group A streptococcus quantum dot immunochromatography test reagent card |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201420217173.4U CN203870107U (en) | 2014-04-29 | 2014-04-29 | Group A streptococcus quantum dot immunochromatography test reagent card |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN203870107U true CN203870107U (en) | 2014-10-08 |
Family
ID=51651047
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201420217173.4U Expired - Lifetime CN203870107U (en) | 2014-04-29 | 2014-04-29 | Group A streptococcus quantum dot immunochromatography test reagent card |
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| Country | Link |
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| CN (1) | CN203870107U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103926403A (en) * | 2014-04-29 | 2014-07-16 | 广州市微米生物科技有限公司 | Streptococcus-A quantum dot immunochromatographic detection reagent card and preparation method thereof |
| CN110568187A (en) * | 2019-08-21 | 2019-12-13 | 江苏硕世生物科技股份有限公司 | Test strip and kit for detecting group A streptococcus antigen and preparation method thereof |
-
2014
- 2014-04-29 CN CN201420217173.4U patent/CN203870107U/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103926403A (en) * | 2014-04-29 | 2014-07-16 | 广州市微米生物科技有限公司 | Streptococcus-A quantum dot immunochromatographic detection reagent card and preparation method thereof |
| CN103926403B (en) * | 2014-04-29 | 2016-01-13 | 广州市微米生物科技有限公司 | A race streptococcus quantum dot immune chromatography detects reagent card and preparation method thereof |
| CN110568187A (en) * | 2019-08-21 | 2019-12-13 | 江苏硕世生物科技股份有限公司 | Test strip and kit for detecting group A streptococcus antigen and preparation method thereof |
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