CN201464476U - System detecting antibody of schistosomiasis through dot immuno-gold filtration assay - Google Patents
System detecting antibody of schistosomiasis through dot immuno-gold filtration assay Download PDFInfo
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- CN201464476U CN201464476U CN2009200758733U CN200920075873U CN201464476U CN 201464476 U CN201464476 U CN 201464476U CN 2009200758733 U CN2009200758733 U CN 2009200758733U CN 200920075873 U CN200920075873 U CN 200920075873U CN 201464476 U CN201464476 U CN 201464476U
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- 102000004965 antibodies Human genes 0.000 title claims abstract description 28
- 108090001123 antibodies Proteins 0.000 title claims abstract description 28
- 239000010931 gold Substances 0.000 title claims abstract description 13
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 13
- 238000004166 bioassay Methods 0.000 title abstract description 3
- 238000001914 filtration Methods 0.000 title abstract 2
- 201000004409 schistosomiasis Diseases 0.000 title abstract 2
- 238000006243 chemical reaction Methods 0.000 claims abstract description 52
- 239000000463 material Substances 0.000 claims abstract description 30
- 239000000427 antigen Substances 0.000 claims abstract description 20
- 102000038129 antigens Human genes 0.000 claims abstract description 20
- 108091007172 antigens Proteins 0.000 claims abstract description 20
- 239000002250 absorbent Substances 0.000 claims abstract description 17
- 230000002745 absorbent Effects 0.000 claims abstract description 17
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 12
- 235000013601 eggs Nutrition 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 6
- 241000237858 Gastropoda Species 0.000 claims description 30
- 206010037660 Pyrexia Diseases 0.000 claims description 30
- 238000005325 percolation Methods 0.000 claims description 26
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 13
- 239000000020 Nitrocellulose Substances 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 7
- 229920001220 nitrocellulos Polymers 0.000 claims description 7
- 210000000987 Immune System Anatomy 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 8
- 230000000694 effects Effects 0.000 abstract description 8
- 238000005406 washing Methods 0.000 abstract description 4
- 239000003795 chemical substances by application Substances 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract 6
- 238000010521 absorption reaction Methods 0.000 abstract 5
- 238000007789 sealing Methods 0.000 abstract 1
- 210000002966 Serum Anatomy 0.000 description 20
- 238000001764 infiltration Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 230000036039 immunity Effects 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000012466 permeate Substances 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 241000668709 Dipterocarpus costatus Species 0.000 description 1
- 241001442514 Schistosomatidae Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000001809 detectable Effects 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 230000035931 haemagglutination Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000003547 immunosorbent Substances 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006011 modification reaction Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 210000001519 tissues Anatomy 0.000 description 1
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Abstract
The utility model relates to a system detecting the antibody of schistosomiasis through dot immuno-gold filtration assay. The system comprises a reaction device, wherein, the reaction device comprises a box body, a box cover, water absorption material and a reaction film; the water absorption material is arranged in the box body; the reaction film is placed on the water absorption material; the box cover is covered on the box body and presses the reaction film; a reaction hole is formed on the box cover, so that part of the reaction film is exposed through the reaction hole; and the reaction hole is an elliptical reaction hole. Preferably, the volume of the elliptical reaction hole is 50 Mul, and the bottom area thereof is 0.5cm2; the elliptical reaction hole is formed in the center of the box cover; the reaction film adopts a microporous film; the thickness of the water absorption material is three mm to five mm; the water absorption material adopts a water absorbent paper; a soluble egg antigen film, a human IgG film and a sealing agent layer are coated on the reaction film in the reaction hole; and the system further comprises a first reagent bottle containing washing liquid and a second reagent bottle containing color development solution. The utility model has the advantages of smart design, simple and convenient use, reduced cost, fast and sensitive detection, and accurate and reliable detection effect, and is suitable for being applied to the field with large scale.
Description
Technical field
The utility model relates to antibody test systems technology field, and particularly snail fever antibody test systems technology field specifically is meant the golden percolation snail fever of a kind of spot immune antibody test system.
Background technology
The snail fever amynologic diagnostic method is very reliable auxiliary diagnosis foundation, also is the most frequently used instrument of epidemiology survey.At present commonly used euzymelinked immunosorbent assay (ELISA), indirect hemagglutination method, DDIA, colloidal gold dot immunity percolation method arranged.Though these methods cut both ways, not obviously difference aspect susceptibility that detects in snail fever and the specificity, all included People's Republic of China's health industry standard in---schistosomiasis diagnosis standard (WS261-2006).Colloidal gold dot immunity percolation method has been an emerging method on China snail fever immunodiagnosis market since the nineties in last century, it is advantageous that easy and simple to handle, reaction fast, do not need any Special Equipment, do not need staff's Special Training, can make the quick diagnosis instrument.But, percolation also has some shortcomings, as, percolation to the requirement of serum than higher, generally require fresh serum, if outmoded serum then need centrifuging, cause reaction film to stop up otherwise infiltration rate will be descended, influence reaction result, and the serum amount that percolation needs is bigger, common 50ul～100ul, these factors all limit its on-the-spot large-scale application.
Therefore, need provide the golden percolation snail fever of a kind of spot immune antibody test system, it is easy to use, and cost reduces, and detects rapid sensitive, detects effect accurately and reliably, is suitable for on-the-spot large-scale application.
The utility model content
The purpose of this utility model is to have overcome above-mentioned shortcoming of the prior art, and the golden percolation snail fever of a kind of spot immune antibody test system is provided, and this detection system design is ingenious, easy to use, cost reduces, and detects rapid sensitive, detect effect accurately and reliably, be suitable for on-the-spot large-scale application.
To achieve these goals, there is following formation in the golden percolation snail fever antibody test of spot immune of the present utility model system:
This spot immune gold percolation snail fever antibody test system, be characterized in, comprise reaction unit, described reaction unit comprises box body, lid, absorbent material and reaction film, and described absorbent material is arranged in described box body, and described reaction film places on the described absorbent material, described lid covers pushes down described reaction film on described box body, have reacting hole on the described lid, the described reaction film of exposed part, described reacting hole are oval reacting hole.
Preferably, the volume of described oval reacting hole is 50 μ l, and floorage is 0.5cm
2
Preferably, described oval reacting hole is arranged on the central authorities of described lid.
Preferably, described reaction film is a microporous barrier.
More preferably, described microporous barrier is a nitrocellulose membrane, and the aperture of described nitrocellulose membrane is 0.45 μ m.
Preferably, the thickness of described absorbent material is 3mm～5mm.
Preferably, described absorbent material is the suction paper washer.
More preferably, described suction paper washer is 32 layers of napkin paper.
Preferably, bag is by soluble egg antigen rete, human IgG rete and sealer layer on the described reaction film in the described reacting hole, and described soluble egg antigen rete and described human IgG rete are arranged in described sealer layer and are separated by described sealer layer.
Preferably, the golden percolation snail fever antibody test of described spot immune system also comprises first reagent bottle that holds cleansing solution and second reagent bottle that holds colour developing liquid.
The beneficial effects of the utility model are specific as follows:
1. the utility model is designed to oval reacting hole with reacting hole, designs ingeniously, has enlarged the serum infiltrating area, be convenient to the abundant washing of cleansing solution, keep filter opening unobstructed, make outmoded serum also can permeate reaction film, save the step of using hydro-extractor to carry out centrifuging, easy to use;
2. of the present utility model antigen coated in the centre of reacting hole, serum adds from central authorities during detection, directly is added in fully contact with it above the antigen, and the serum consumption reduces (when volume is 50 μ l at double, adopt oval reacting hole can reduce serum consumption 25 μ l), cost reduces greatly;
3. absorbent material of the present utility model adopts the suction paper washer, and the suitableeest thickness is 32 layers of napkin paper, has improved infiltration rate; By adjusting the gold mark two anti-concentration that reach envelope antigen, the suitableeest gold mark two anti-concentration are A520nm 1.0, and antigen concentration is 0.2mg/ml, has strengthened reaction sensitivity; By adjusting gold mark cleansing solution prescription, the optimum concentration of Tween-20 is 2%, and background is more clear, detects rapid sensitive, detects effect accurately and reliably;
4. the utility model is not done requirement to serum purity, and outmoded serum need not centrifugal can the use; Reaction system serum consumption is few, only needs 25 μ l; Reaction rate is fast, 1 minute overall process time spent half; Detect effect accurately and reliably, be suitable for on-the-spot large-scale application.
Description of drawings
Fig. 1 is the structural representation of a specific embodiment of the present utility model.
Fig. 2 is the decomposing schematic representation of the reaction unit of specific embodiment shown in Figure 1.
Fig. 3 a-c is the testing result synoptic diagram that adopts the utility model to detect.
Embodiment
In order more to be expressly understood technology contents of the present utility model, describe in detail especially exemplified by following examples.
See also shown in Fig. 1-2, the golden percolation snail fever antibody test of spot immune of the present utility model system comprises reaction unit 1, described reaction unit 1 comprises box body 11, lid 12, absorbent material 13 and reaction film 14, described absorbent material 13 is arranged in described box body 11, described reaction film 14 places on the described absorbent material 13, described lid 12 covers pushes down described reaction film 14 on described box body 11, have reacting hole 15 on the described lid 12, the described reaction film 14 of exposed part, described reacting hole 15 is oval reacting hole.Therefore, constancy of volume, reacting hole 15 is designed to ellipse, has enlarged the serum infiltrating area, is convenient to the abundant washing of cleansing solution to reaction film 14, keeps filter opening unobstructed, makes outmoded serum also can permeate, and has save the step of using hydro-extractor to carry out centrifuging.
In specific embodiment of the utility model, the volume of described oval reacting hole is 50 μ l, and floorage is 0.5cm2.
In specific embodiment of the utility model, described oval reacting hole is arranged on the central authorities of described lid 12.
Preferably, described reaction film 14 is microporous barriers.In specific embodiment of the utility model, described microporous barrier is a nitrocellulose membrane, and the aperture of described nitrocellulose membrane is 0.45 μ m.
Preferably, the thickness of described absorbent material 13 is 3mm～5mm.
Preferably, described absorbent material 13 is suction paper washers. by adjusting the thickness of reaction film 14 following suction paper washers, improve infiltration rate. the commercially available napkin paper (suction paper washer) of the different numbers of plies is lining in reaction film 14 times, determine the suitableeest number of plies according to detecting effect. in specific embodiment of the utility model, described suction paper washer is 32 layers of napkin paper.
Preferably, bag is by soluble egg antigen rete, human IgG rete and sealer layer on the described reaction film 14 in the described reacting hole 15, and described soluble egg antigen rete and described human IgG rete are arranged in described sealer layer and are separated by described sealer layer.In specific embodiment of the utility model, the soluble egg antigen rete is positioned at the centre of reacting hole 15, and the other drop human IgG of antigen is as the Quality Control point.Serum adds from central authorities during detection, directly is added in fully contact with it above the antigen, and the serum consumption can reduce (25 μ l) at double.
Preferably, the golden percolation snail fever antibody test of described spot immune system also comprises first reagent bottle 2 that holds cleansing solution and second reagent bottle 3 that holds colour developing liquid.Colour developing liquid is gold mark two and resists, and adjusts the gold mark two anti-concentration that reach envelope antigen, but intensified response sensitivity.In specific embodiment of the utility model, envelope antigen adopts homemade blood fluke soluble egg antigen, and concentration is 0.2mg/ml; " snail fever antibody diagnosing reagent country the reference material " (lot number: that adopts Nat'l Pharmaceutical ﹠ Biological Products Control Institute to provide 20041108) as the suitableeest antigen concentration and gold mark two anti-checking serum; Washing agent is formulated as 0.02mol/LpH8.0TBS substantially, adds the Tween-20 of variable concentrations (0.5%～5%) then, selects optimum concentration according to the background sharpness, and the optimum concentration of Tween-20 is 2%, and background is more clear.
Manufacture process of the present utility model is: fill up suction bedding and padding 13 in the plastic box body 11 of 4.0cm * 2.2cm, lid 12 central authorities open an area 0.5cm
2Oval-shaped reacting hole 15, be close to the suction bedding and padding under the hole and place a small pieces nitrocellulose membrane (aperture 0.45 μ m), film central authorities drop 1 μ l soluble egg antigen (with 0.02mol/L PBS dilution), the other drop 0.5 μ l human IgG of antigen is as the Quality Control point, natural drying at room temperature 2h, drip sealer, the dry airtight 4 ℃ of preservations in back are standby.
In the oval-shaped reacting hole 15 of the central authorities of reaction unit 1, drip 2 cleansing solutions during detection, infiltrate the back and add 25 μ l in reacting hole 15 central authorities and examined serum, treat that liquid fully sucks, add 2 cleansing solutions, it is anti-to add 4 gold marks two after the infiltration, adds 2 cleansing solutions at last, gets final product judged result after the infiltration.Shown in Fig. 3 a-c, it is positive to occur two punctations in the reacting hole 15, and it is negative a punctation to occur, and the immaculate ecbatic is invalid.
Adopt the test result of the utility model to snail fever antibody diagnosing reagent country reference material, the detection coincidence rate that demonstrates the yin and yang attribute reference material is 100%, sensitivity is 1:4, precision test 10 times is all positive, the colour developing uniformity all meets the detection quality requirements (seeing the following form 1) of national reference material.Participation is gone through primary dcreening operation and formal 2 stages of test and appraisal by the Chinese snail fever immune diagnostic reagent laboratory test and appraisal work of snail fever Expert Advisory Committee (EAC) of the Ministry of Public Health in tissue in 2008.The susceptibility 92% of kit, specificity 95.08%, Youden index 0.87, kappa value 0.87 has better repeatability.
The test result of table 1 snail fever antibody diagnosing reagent country reference material
| Serum Sera | Umber No.sera | Negative (number) No.negative | Positive (number) No.positive | Coincidence rate (%) Coincident rates |
| Negative reference material Negative reference sera | 15 | 15 | 0 | 100 |
| Serum Sera | Umber No.sera | Negative (number) No.negative | Positive (number) No.positive | Coincidence rate (%) Coincident rates |
| Positive reference material Positive reference sera | 15 | 0 | 15 | 100 |
| Sensitivity reference material Detectable minimum reference sera | 4 (1:2～1:16) | ＞1:8 | ≤1:4 | |
| Precision reference material Precision reference sera | 10 | 0 | 10 |
To sum up, spot immune gold percolation snail fever antibody test system design of the present utility model is ingenious, easy to use, and cost reduces, and detects rapid sensitive, detects effect accurately and reliably, is suitable for on-the-spot large-scale application.
In this instructions, the utility model is described with reference to its certain embodiments.But, still can make various modifications and conversion obviously and not deviate from spirit and scope of the present utility model.Therefore, instructions and accompanying drawing are regarded in an illustrative, rather than a restrictive.
Claims (10)
1. spot immune gold percolation snail fever antibody test system, it is characterized in that, comprise reaction unit, described reaction unit comprises box body, lid, absorbent material and reaction film, and described absorbent material is arranged in described box body, and described reaction film places on the described absorbent material, described lid covers pushes down described reaction film on described box body, have reacting hole on the described lid, the described reaction film of exposed part, described reacting hole are oval reacting hole.
2. the golden percolation snail fever of spot immune according to claim 1 antibody test system is characterized in that the volume of described oval reacting hole is 50 μ l, and floorage is 0.5cm
2
3. the golden percolation snail fever of spot immune according to claim 1 antibody test system is characterized in that described oval reacting hole is arranged on the central authorities of described lid.
4. the golden percolation snail fever of spot immune according to claim 1 antibody test system is characterized in that described reaction film is a microporous barrier.
5. the golden percolation snail fever of spot immune according to claim 4 antibody test system is characterized in that described microporous barrier is a nitrocellulose membrane, and the aperture of described nitrocellulose membrane is 0.45 μ m.
6. the golden percolation snail fever of spot immune according to claim 1 antibody test system is characterized in that the thickness of described absorbent material is 3mm～5mm.
7. the golden percolation snail fever of spot immune according to claim 1 antibody test system is characterized in that described absorbent material is the suction paper washer.
8. the golden percolation snail fever of spot immune according to claim 7 antibody test system is characterized in that described suction paper washer is 32 layers of napkin paper.
9. the golden percolation snail fever of spot immune according to claim 1 antibody test system, it is characterized in that, bag is by soluble egg antigen rete, human IgG rete and sealer layer on the described reaction film in the described reacting hole, and described soluble egg antigen rete and described human IgG rete are arranged in described sealer layer and are separated by described sealer layer.
10. the golden percolation snail fever of spot immune according to claim 1 antibody test system, it is characterized in that the golden percolation snail fever antibody test of described spot immune system also comprises first reagent bottle that holds cleansing solution and holds second reagent bottle of colour developing liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009200758733U CN201464476U (en) | 2009-08-05 | 2009-08-05 | System detecting antibody of schistosomiasis through dot immuno-gold filtration assay |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009200758733U CN201464476U (en) | 2009-08-05 | 2009-08-05 | System detecting antibody of schistosomiasis through dot immuno-gold filtration assay |
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| CN201464476U true CN201464476U (en) | 2010-05-12 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012068709A1 (en) * | 2010-11-25 | 2012-05-31 | 西安微通生物技术有限公司 | Kit of dot immunogold orientated filtration assay and use thereof |
| CN103076447A (en) * | 2012-12-27 | 2013-05-01 | 上海市疾病预防控制中心 | Schistosoma egg crude antigen purification method, related purified antigen and schistosoma antibody detection colloidal gold immunoassay kit |
| CN103076455A (en) * | 2012-12-26 | 2013-05-01 | 上海奥普生物医药有限公司 | Kit for quickly and quantificationally detecting serum amyloid A, and preparation and application thereof |
| CN105785040A (en) * | 2016-03-31 | 2016-07-20 | 新疆农业大学 | EHV-1 serum antibody detection kit as well as preparation method and application |
-
2009
- 2009-08-05 CN CN2009200758733U patent/CN201464476U/en not_active Expired - Lifetime
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012068709A1 (en) * | 2010-11-25 | 2012-05-31 | 西安微通生物技术有限公司 | Kit of dot immunogold orientated filtration assay and use thereof |
| CN103076455A (en) * | 2012-12-26 | 2013-05-01 | 上海奥普生物医药有限公司 | Kit for quickly and quantificationally detecting serum amyloid A, and preparation and application thereof |
| CN103076447A (en) * | 2012-12-27 | 2013-05-01 | 上海市疾病预防控制中心 | Schistosoma egg crude antigen purification method, related purified antigen and schistosoma antibody detection colloidal gold immunoassay kit |
| CN103076447B (en) * | 2012-12-27 | 2015-04-08 | 上海市疾病预防控制中心 | Schistosoma egg crude antigen purification method, related purified antigen and schistosoma antibody detection colloidal gold immunoassay kit |
| CN105785040A (en) * | 2016-03-31 | 2016-07-20 | 新疆农业大学 | EHV-1 serum antibody detection kit as well as preparation method and application |
| CN105785040B (en) * | 2016-03-31 | 2018-05-25 | 新疆农业大学 | A kind of EHV-1 Serum Antibody Detections kit and preparation method and application |
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Legal Events
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 200336 No. 1380 West Zhongshan Road, Shanghai Patentee after: Shanghai Disease Prevention and Control Centre Patentee after: Shanghai Shen Kai Biological Technology Co., Ltd. Address before: 200336 No. 1380 West Zhongshan Road, Shanghai Patentee before: Shanghai Disease Prevention and Control Centre Patentee before: Shanghai Jikong Biotechnology Co., Ltd. |
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| CX01 | Expiry of patent term |
Granted publication date: 20100512 |
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| CX01 | Expiry of patent term |