CN204228713U - For the test card that DDi detects - Google Patents
For the test card that DDi detects Download PDFInfo
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- CN204228713U CN204228713U CN201420674146.XU CN201420674146U CN204228713U CN 204228713 U CN204228713 U CN 204228713U CN 201420674146 U CN201420674146 U CN 201420674146U CN 204228713 U CN204228713 U CN 204228713U
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Abstract
The utility model provides a kind of test card, and it comprises detector bar and shell, and described detector bar comprises with lower part: sample is in conjunction with bar, and described sample is in conjunction with first antibody bar with the anti-DDi marked by the fluorescent microsphere of two kinds of different-grain diameter sizes; Bag is by bar, and described bag bar is had detection zone and quality control band, and described detection zone has the second antibody of anti-DDi, and described quality control band is fixed with the antibody of against murine IgG; With water suction bar; Described shell comprises with lower part: Ka Gai, described card covers and is provided with well and detect aperture, described well is positioned at the upside of described sample in conjunction with bar to expose the subregion of described sample in conjunction with bar, and described detect aperture is positioned at described bag by the upside of bar to expose the Zone Full of described detection zone and described quality control band; And base.Test card detection sensitivity of the present utility model is high, and the range of linearity is wide and structure simple, is of value to clinical practice.
Description
Technical field
The utility model relates to a kind of test card for DDi immune detection.
Background technology
Current clinical conventional method comprises: latex enhancing immune turbidimetry, euzymelinked immunosorbent assay (ELISA), chemoluminescence method, colloidal gold immunity chromatography, fluorescence immune chromatography method etc.Though latex enhancing immune turbidimetry realizes high flux screening, the detection sensitivity needed for some determinand requires high, and when causing detecting, low value sample CV value is excessive.Though euzymelinked immunosorbent assay (ELISA) and chemoluminescence method can be quantitatively accurate, but consuming time long for its detection of judgement aspect of some disease, greatly delay the treatment that may need.Colloidal gold immunity chromatography is simple to operate, but can only realize sxemiquantitative.By contrast, fluorescence immune chromatography method both can realize quick detection, can carry out accurate quantification again, for clinical practice brings great convenience.
But there is certain defect in the fluorescence immune chromatography method using fluorescent microsphere as label conventional at present in application.The range of linearity that Large stone fluorescent microsphere detects is inadequate, and therefore high level detects inaccurate.The measured value of the low side that small particle diameter fluorescent microsphere detects is inaccurate, and CV value is excessive.In addition, at present the detector bar developed based on above-mentioned detection method and/or test card are also existed to complex process, cost are high, the problems such as sensitivity and poor accuracy.
Utility model content
technical matters
The test card that can realize accurately detecting DDi immune detection is fast lacked in currently available technology.Therefore, need to develop a kind of test card with highly sensitive DDi immune detection.
technical scheme
On the one hand, the utility model provides a kind of test card for DDi immune detection, it is characterized in that, described test card comprises detector bar and the shell with coated described detector bar,
Wherein, described detector bar comprise to each other order overlap joint with lower part:
1) there is the first antibody sample of the anti-DDi marked by the fluorescent microsphere of two kinds of different-grain diameter sizes in conjunction with bar,
2) there is the bag of detection zone and quality control band by bar, wherein,
2a) described detection zone has the second antibody of anti-DDi,
2b) described quality control band is fixed with the antibody of against murine IgG, and
3) absorb water bar;
Wherein, described shell comprise mutually fasten with lower part:
4) Ka Gai of well and detect aperture is provided with, wherein
4a) described well is positioned at the upside of described sample in conjunction with bar, and its size is enough to expose the subregion of described sample in conjunction with bar,
4b) described detect aperture is positioned at described bag by the upside of bar, and its size is enough to the Zone Full exposing described detection zone and described quality control band, and
5) base.
In one embodiment, described detector bar comprise further for arrange sample in conjunction with bar, wrap by bar and water suction bar base plate.
In one embodiment, the particle size scope of the fluorescent microsphere of described two kinds of different-grain diameter sizes is between 100nm to 400nm
In one embodiment, the particle size of the fluorescent microsphere of described two kinds of different-grain diameter sizes is respectively 100nm and 300nm.
In one embodiment, be interposed between 0.3cm to 0.5cm between described detection zone and quality control band.
In one embodiment, the different epitopes of the first antibody of described anti-DDi and the second antibody difference specific binding DDi of described anti-DDi.
In one embodiment, described sample is prepared by glass fibre element film or polyester film in conjunction with bar.
In one embodiment, described bag is nitrocellulose filter by bar.
beneficial effect
Compared with other DDi immune detection goods of prior art, DDi immune detection card of the present utility model has the following advantages:
The design of the fluorescent microsphere of two kinds of different-grain diameters can realize high sensitivity and the wide detection range of linearity simultaneously;
The employing of double antibody sandwich method further increases detection sensitivity;
The structure of test card makes it can be used in corresponding fluorescence detector, to meet the needs that clinical extensive sample detection is analyzed; With
Test card structure is simple, is of value to the simplification of the technological process of production and reduces production cost, can reduce the expense of clinical detection thus.
Accompanying drawing explanation
The schematic diagram of Fig. 1 test card for DDi immune detection of the present utility model.
The schematic diagram of detector bar in Fig. 2 the utility model test card.
Reference numeral:
1 test card 2-1 base plate 2-7 Large stone fluorescent microsphere
2 detector bar 2-2 samples are in conjunction with bar 2-8 small particle diameter fluorescent microsphere
3 card lid 2-3 bags are by bar
4 base 2-4 water suction bars
5 well 2-5 detection zone
6 detect aperture 2-6 quality control bands
Embodiment
Hereinafter come with reference to the accompanying drawings to describe illustrative embodiments in more detail.Described accompanying drawing is used for illustrating the utility model, but not is limited.
Fig. 1 is the schematic diagram of the test card for DDi immune detection of the present utility model.
As shown in Figure 1, DDi immune detection card 1 of the present utility model comprises detector bar 2 and shell, and shell comprises card lid 3 and base 4, card lid 3 is provided with well 5 and detect aperture 6.Detector bar 2 is placed in enclosure by linking closely each other by card lid 3 and base 4.On card lid 3, the large I of well 5 to expose on detector bar 2 sample in conjunction with the subregion of bar, and the large I of detect aperture 6 exposes Zone Full detector bar 2 wrapped by detection zone in bar and quality control band.The structure of detector bar 2 is specified in Fig. 2.
Card lid 3 and base 4 all can be made up of such as plastic or other material.Well 5 and detect aperture 6 can be square, rectangle, trapezoidal, oval, circular, etc. shape.The lengthwise of well 5 can be 0.05cm to 0.6cm, preferred 0.1cm to 0.5cm.Growing crosswise of well 5 can be 0.15cm to 0.45cm, preferred 0.2cm to 0.4cm.The lengthwise of detect aperture 6 can be 0.8cm to 2.5cm, preferred 1.0cm to 2.3cm.Growing crosswise of detect aperture 6 can be 1.2cm to 2.1cm, preferred 1.5cm to 1.9cm.
Test card of the present utility model makes DDi immune detection can carry out in corresponding fluorescence detector, can realize large batch of sample thus and detect fast, have very important clinical meaning.
Fig. 2 is the schematic diagram for detector bar in the test card of DDi immune detection of the present utility model.
As previously mentioned, detector bar 2 is passed through by card lid 3 and base 4 enclosure being closely buckled in test card 1 each other.As shown in Figure 2, detector bar 2 comprise sample in conjunction with bar 2-2, wrap by bar 2-3 and water suction bar 2-4, wherein in conjunction with bar 2-2, wrap and be pasted onto on base plate 2-1 by overlap joint successively by bar 2-3 and water suction bar 2-4.
Sample can play reception and the effect in conjunction with sample in conjunction with bar 2-2 simultaneously.Adopt sample strip and the method in conjunction with bar independent design compared to tradition, the detector bar in the utility model test card adopts sample strip and the design in conjunction with bar integration, thus enormously simplify production technology and flow process.In one embodiment, sample is prepared by glass fibre element film or polyester film in conjunction with bar 2-2.
Sample is in conjunction with antibody bar 2-2 being coated with further a kind of anti-DDi that the different fluorescent microsphere of particle size marks.The particle size scope of Large stone fluorescent microsphere 2-7 is between 300nm to 500nm.In one embodiment, the particle size of Large stone fluorescent microsphere 2-7 is 300nm.The particle size scope of small particle diameter fluorescent microsphere 2-8 is between 50nm to 150nm.In one embodiment, the particle size of small particle diameter fluorescent microsphere 2-8 is 100nm.The antibody that Large stone fluorescent microsphere 2-7 marks is conducive to the detection of low value to reach the requirement of sensitivity, and the antibody that small particle diameter fluorescent microsphere 2-8 marks makes the range of linearity detected can cover high level region.Therefore, detector bar of the present utility model can meet the requirement of high sensitivity and the wide detection range of linearity.
Bag bar 2-3 is had detection zone 2-5 and quality control band 2-6.Detection zone 2-5 has the antibody of another kind of anti-DDi.With sample in conjunction with compared with the antibody on bar 2-2, the epitope of the DDi that the antibody specific binding on detection zone 2-5 is different.Quality control band 2-6 is fixed with the antibody of against murine IgG, and it is used as Quality Control and contrast.Be interposed between between detection zone and quality control band between 0.2cm to 0.6cm, between preferred 0.3cm to 0.5cm.Bag can be made up of materials such as such as nitrocellulose filters by bar 2-3.
Water suction bar 2-4 is overlapped by bar 2-3 with bag.Water suction bar 2-4 can be made up of the such as material such as velveteen, glass fibre.
Embodiment
Following examples are for illustration of the utility model.
In following embodiment, described DDi first antibody, D-mono-aggressiveness second antibody, dynamics, fluorescent microsphere, 2-(N-morpholine) ethyl sulfonic acid (MES), carbodiimide (EDC), N-hydroxy-succinamide (NHS), bovine serum albumin(BSA) (BSA), Tween-20, trehalose, glass fibre element film, polyester film, nitrocellulose filter, high-intensity water absorbent paper are commercially available prod.
The preparation method that described application two kinds of different-grain diameter fluorescent microspheres detect the fluorescence immune chromatography detector bar of DDi is:
1) sample is in conjunction with the preparation of bar
A, sample are in conjunction with the pre-service of bar
Glass fibre element film or polyester film are put into sample in conjunction with the bar treating fluid (0.1M containing 1%BSA and 0.1%Tween-20, boric acid-the borate buffer solution of pH8.0-8.6 or the 0.05M containing 1%BSA and 0.1%Tween-20, the Tris-HCl damping fluid of pH7.4-8.2) middle after immersion treatment 3-5 hour, in 35-39 DEG C of air dry oven, dry 3-5 hour.
The preparation of two kinds of different-grain diameter fluorescent microspheres of B, labelled antibody
By DDi first antibody with boric acid-borate buffer solution 4 DEG C of dialysed overnight of 0.1M, pH8.0-8.6, adjustment concentration is 2mg/ml-5mg/ml.
Use 0.05M, the small particle diameter fluorescent microsphere of 2-(N-morpholine) ethyl sulfonic acid (MES) the activation buffer washing 50-150nm of pH4.5-5.0, adding carbodiimide (EDC) and N-hydroxy-succinamide (NHS) makes the two final concentration be 15-60mmol, room temperature reaction 20-30 minute, abundant washing fluorescent microsphere, with 0.05M, the first antibody of dialysing is added after boric acid-borate buffer solution redissolution of pH8.0-8.6, the mass ratio of first antibody and fluorescent microsphere is made to be 1: 8 to 1: 60, room temperature reaction is after 2 hours, add the 0.02M containing 10%BSA, the phosphate buffer room temperature of pH7.2-7.6 closes 30 minutes, abundant washing fluorescent microsphere, with the 0.05M containing 1%BSA and 0.1%Tween-20, the conserving liquid of the Tris-HCl damping fluid of pH7.4-8.2 redissolves to original volume.
Use 0.05M, the Large stone fluorescent microsphere of 2-(N-morpholine) ethyl sulfonic acid (MES) the activation buffer washing 300-500nm of pH4.5-5.0, adding carbodiimide (EDC) and N-hydroxy-succinamide (NHS) makes the two final concentration be 15-60mmol, room temperature reaction 20-30 minute, abundant washing fluorescent microsphere, with 0.05M, the first antibody of dialysing is added after boric acid-borate buffer solution redissolution of pH8.0-8.6, the mass ratio of first antibody and fluorescent microsphere is made to be 1: 20 to 1: 80, room temperature reaction is after 2 hours, add the 0.02M containing 10%BSA, the phosphate buffer room temperature of pH7.2-7.6 closes 30 minutes, abundant washing fluorescent microsphere, with the 0.05M containing 1%BSA and 0.1%Tween-20, the conserving liquid of the Tris-HCl damping fluid of pH7.4-8.2 redissolves to original volume.
By the fluorescent microsphere of two kinds of different-grain diameters of above-mentioned mark first antibody in 1: 10 to 10: 1 ratio mix, namely prepare two kinds of different-grain diameter fluorescent microspheres of labelled antibody.
C, sample are in conjunction with the preparation of bar
Two kinds of different-grain diameter fluorescent microsphere potpourris of the labelled antibody prepared by step B use and quantitatively spray film instrument with on the plain film of the glass fibre that 2 μ l/cm-8 μ l/cm even application are pretreated in above-mentioned steps A or polyester film, in 35-39 DEG C of air dry oven, dry 2-4 hour, add drying agent and seal up for safekeeping for subsequent use.
2) bag is by the preparation of bar
The preparation of A, detection zone: by DDi second antibody with 0.01-0.02M, Tris-HCl damping fluid 4 DEG C of dialysed overnight of pH7.4-8.2, use the coating buffer (0.01-0.02M containing 0.1-10% trehalose, the Tris-HCl damping fluid of pH7.4-8.2) the DDi second antibody after dialysis is diluted to the concentration of 0.2mg/ml-4mg/ml, use quantitatively spray film instrument with 1.2 μ 1/cm even application on nitrocellulose filter, in 35-39 DEG C of air dry oven, dry 2-4 hour.
The preparation of B, quality control band: use the coating buffer (0.01-0.02M containing 0.1-10% trehalose, the Tris-HCl damping fluid of pH7.4-8.2) anti-mouse IgG is diluted to the concentration of 0.5mg/ml-3mg/ml, use quantitatively spray film instrument with 0.8 μ l/cm even application on nitrocellulose filter, in 35-39 DEG C of air dry oven, dry 2-4 hour.
C, wrap by the preparation of bar: with the confining liquid (0.01-0.02M containing 1-10%BSA and 1-10% trehalose, the Tris-HCl damping fluid of pH7.4-8.2) by the cellulose nitrate membrane closure 1-2 hour containing detection zone and quality control band, take out in rearmounted 35-39 DEG C air dry oven and dry 2-4 hour, envelope is for subsequent use.
3) assembling of detector bar
Be less than 30% in humidity, under temperature 20-30 DEG C of condition, carry out the assembling of detector bar.Base plate selects PVC material, connect in turn in the direction of base plate the same face and the partly overlapping sample pad of adjacent regions, wrap by bar and water suction bar, the overlap length of overlapping region is 1-3mm.When bag is covered base plate by bar, the position of detection zone is near sample in conjunction with bar, and the position of quality control band is near water suction bar.Water suction bar is high-intensity water absorbent paper.Carry out cutting after assembling, forming width is the detector bar of 0.2cm-1cm.
4) assembling of test card
Be less than 30% in humidity, under temperature 20-30 DEG C of condition, carry out the assembling of test card.Above-mentioned detector bar is placed in the centre position of plastic feet, the base plate of detector bar is connected with plastic feet, and fastens plastic clip lid.Card covers and is provided with well and detect aperture, and well is opened on the upside of sample pad, exposes the subregion of sample pad, and detect aperture is opened on bag by the upside of bar, exposes the Zone Full of detection zone and quality control band.
5) mensuration of calibration object and measuring samples
The standard items of 100 μ 1 or measuring samples are joined the well of test card, carry out immunochromatography reaction, after 10-15 minute, test card is placed in special fluorescence detector, carry out quantitative measurement by the size reading fluorescence signal.
The fluorescence immune chromatography detector bar that application provided by the utility model two kinds of different-grain diameter fluorescent microspheres quantitatively detect DDi has taken into account the range of linearity of low value sensitivity and detection, and quick, single part can be realized quantitatively detect, in addition volume little, be easy to carry about with one and preserve, for Clinical practice provides great convenience.In addition, compare traditional detection DDi containing sample in conjunction with bar, in conjunction with bar, wrap by the fluorescent chromatographic detector bar of bar, water suction bar four parts, detector bar provided by the utility model only contains sample reagent bar, wraps by bar, water suction bar three parts, therefore it simplifies more, preparation method is simpler, batch production can be realized, be conducive to large-scale promotion application, possess wide market outlook.
Above embodiment is only used to done citing is clearly described, the restriction not to embodiment.For those of ordinary skill in the field, make other improvement on the basis of the above description to be still within protection domain of the present utility model.
Claims (8)
1. for the test card that DDi detects, it is characterized in that, described test card comprises detector bar and the shell with coated described detector bar,
Wherein, described detector bar comprise to each other order overlap joint with lower part:
1) there is the first antibody sample of the anti-DDi marked by the fluorescent microsphere of two kinds of different-grain diameter sizes in conjunction with bar,
2) there is the bag of detection zone and quality control band by bar, wherein,
2a) described detection zone has the second antibody of anti-DDi,
2b) described quality control band is fixed with the antibody of against murine IgG, and
3) absorb water bar;
Wherein, described shell comprise mutually fasten with lower part:
4) Ka Gai of well and detect aperture is provided with, wherein
4a) described well is positioned at the upside of described sample in conjunction with bar, and its size is enough to expose the subregion of described sample in conjunction with bar,
4b) described detect aperture is positioned at described bag by the upside of bar, and its size is enough to the Zone Full exposing described detection zone and described quality control band, and
5) base, described detector bar is fastened on inside by wherein said base and described Ka Gai each other.
2. test card according to claim 1, is characterized in that, described detector bar comprise further for arrange sample in conjunction with bar, wrap by bar and water suction bar base plate.
3. test card according to claim 1, is characterized in that, the particle size scope of the fluorescent microsphere of described two kinds of different-grain diameter sizes is between 100nm to 400nm.
4. test card according to claim 1, is characterized in that, the particle size of the fluorescent microsphere of described two kinds of different-grain diameter sizes is respectively 100nm and 300nm.
5. test card according to claim 1, is characterized in that, is interposed between 0.3cm to 0.5cm between described detection zone and quality control band.
6. test card according to claim 1, is characterized in that, the different epitopes of the first antibody of described anti-DDi and the second antibody difference specific binding DDi of described anti-DDi.
7. test card according to claim 1, is characterized in that, described sample is prepared by glass fibre element film or polyester film in conjunction with bar.
8. test card according to claim 1, is characterized in that, described bag is nitrocellulose filter by bar.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201420674146.XU CN204228713U (en) | 2014-11-13 | 2014-11-13 | For the test card that DDi detects |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201420674146.XU CN204228713U (en) | 2014-11-13 | 2014-11-13 | For the test card that DDi detects |
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| CN204228713U true CN204228713U (en) | 2015-03-25 |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106290822A (en) * | 2016-07-28 | 2017-01-04 | 武汉景川诊断技术股份有限公司 | D dimer immunity latex microsphere preparation method and application |
| CN106405078A (en) * | 2016-08-31 | 2017-02-15 | 中山市创艺生化工程有限公司 | D-dimer immunofluorescent quantitative test strip and preparation method thereof |
| CN107525933A (en) * | 2017-07-21 | 2017-12-29 | 王贤俊 | The measure kit and its application process of a kind of D dimers |
| CN110068679A (en) * | 2018-01-23 | 2019-07-30 | 深圳市易瑞生物技术股份有限公司 | Immuno-chromatography detection device |
| CN110823874A (en) * | 2018-08-13 | 2020-02-21 | 博阳生物科技（上海）有限公司 | Homogeneous phase chemiluminescence detection kit and application thereof |
| CN110823873A (en) * | 2018-08-13 | 2020-02-21 | 博阳生物科技（上海）有限公司 | Chemiluminescence analysis method and application thereof |
| CN116298252A (en) * | 2023-03-07 | 2023-06-23 | 上海赫景医药科技有限公司 | Preparation method and application of novel PET (polyethylene terephthalate) substituted sample pad and binding pad test strip |
-
2014
- 2014-11-13 CN CN201420674146.XU patent/CN204228713U/en active Active
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106290822A (en) * | 2016-07-28 | 2017-01-04 | 武汉景川诊断技术股份有限公司 | D dimer immunity latex microsphere preparation method and application |
| CN106290822B (en) * | 2016-07-28 | 2018-06-19 | 武汉景川诊断技术股份有限公司 | Latex microsphere preparation method and application is immunized in d-dimer |
| CN106405078A (en) * | 2016-08-31 | 2017-02-15 | 中山市创艺生化工程有限公司 | D-dimer immunofluorescent quantitative test strip and preparation method thereof |
| CN107525933A (en) * | 2017-07-21 | 2017-12-29 | 王贤俊 | The measure kit and its application process of a kind of D dimers |
| CN110068679A (en) * | 2018-01-23 | 2019-07-30 | 深圳市易瑞生物技术股份有限公司 | Immuno-chromatography detection device |
| CN110823874A (en) * | 2018-08-13 | 2020-02-21 | 博阳生物科技（上海）有限公司 | Homogeneous phase chemiluminescence detection kit and application thereof |
| CN110823873A (en) * | 2018-08-13 | 2020-02-21 | 博阳生物科技（上海）有限公司 | Chemiluminescence analysis method and application thereof |
| CN116298252A (en) * | 2023-03-07 | 2023-06-23 | 上海赫景医药科技有限公司 | Preparation method and application of novel PET (polyethylene terephthalate) substituted sample pad and binding pad test strip |
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