CN201203618Y - Immune colloidal gold test card for new castle disease virus - Google Patents
Immune colloidal gold test card for new castle disease virus Download PDFInfo
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- CN201203618Y CN201203618Y CNU2008200984369U CN200820098436U CN201203618Y CN 201203618 Y CN201203618 Y CN 201203618Y CN U2008200984369 U CNU2008200984369 U CN U2008200984369U CN 200820098436 U CN200820098436 U CN 200820098436U CN 201203618 Y CN201203618 Y CN 201203618Y
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Abstract
The utility model provides an immune colloidal gold test card for Newcastle disease, which comprises a reagent strip and a test card box, wherein the reagent strip is composed of a back lining, a sample pad, a combined pad, a NC film and an absorbing pad, the middle part of the back lining has a NC film, the two ends of the NC film is respectively connected to the combined pad and the absorbing pad; the sample pad is arranged above the combination pad; the combined pad is sprayed with a gold labeled antibody Ab1; and the NC film is sprayed with a capture antibody Ab2 having a line width of 1 millimeter, and a quality control antibody Ab3. The test card box is composed of a box body and a box cover, the box cover has a sample application hole and a reading window, the middle part of the box has a groove for fixing the reagent strip, the two sides of the groove are evenly provided with convex columns for connecting to the box cover. The immune colloidal gold test card has the characteristics of simple structure, convenient and fast testing operation, and low cost; and is suitable for basic level or clinical detecting Newcastle disease virus. The result can be got in about 10 minutes, and the sensibility reaches to 10 ng/ml.
Description
Technical field
The utility model relates to the technology that units such as plant, epidemic prevention quarantine departments and medical institutions diagnosed or detected Avian pneumo-encephalitis virus, is specifically related to the test card of fast detecting Avian pneumo-encephalitis virus.
Background technology
(Newcastle disease is a kind of acute, sepsis, height contagious disease ND) to Avian pneumo-encephalitis virus, and it is propagated rapidly, the M ﹠ M height.In case outburst can cause immeasurable economic loss to poultry husbandry.Therefore units such as plant, epidemic prevention quarantine departments and medical institutions often need diagnose or testing result.State General Administration for Quality Supervision issued the documents in calendar year 2001, required food meat export enterprise is carried out per two months periodic monitorings once, and this regulation requires with chicken embryo virus separation method.Chicken embryo virus separation method is the special method of a kind of sensitivity, but consuming time longer, needs about 3 weeks approximately, and operates loaded down with trivial detailsly, can only play the monitoring effect, can not make fast detecting usefulness, is unfavorable for speeding passage through customs and controls poultry and carry virus.CN1827775A (number of patent application is 200510008997.6) " detects the nucleotide sequences of Newcastle virus; the kit and the method for inspection ", this invention selects F gene of NDV strain (synthetic F albumen) as target region, selection can reflect that the conserved regions design of cracking site characteristic and shortage secondary structure is many to primer and probe, set up fluorescent PCR and detect Avian pneumo-encephalitis virus, this method than the highly sensitive 100-1000 of traditional PCR method doubly, but need about 4 hours to going out the result from sample preparation, the enforcement of this method also needs expensive instruments such as quantitative real time PCR Instrument, should not promote in basic unit.
At present, prior art mainly comprises the diagnosis or the detection of Avian pneumo-encephalitis virus: blood clotting suppresses (HI) test and detects antihemagglutinin (HA) antibody; Enzyme linked immunosorbent assay (ELISA) (ELISA) method detects Avian pneumo-encephalitis virus total specific antibody or antigen.Though wherein HI tests relatively sensitivity, be subjected to extraneous factor to influence big and weakened its practicality to be worth; ELISA detects the positive serum time early, and the susceptibility height is compared with the HI test and to be more suitable in serosurvey in enormous quantities, but because of its conditional request height, operation inconvenience, failing to bring into play it in actual applications should have effect.And there is false positive (non-specific fluorescence) problem in immunofluorescence technique (IFT).Utilize gene magnification (PCR) method can directly detect the gene of virus, animal test institute of the Ministry of Agriculture at first sets up the strong malicious ewcastle disease of single stage method PT-PCR fast detecting in ewcastle disease test experience chamber at home, and detection time of ewcastle disease was shortened within 12 hours.But required apparatus expensive has been subjected to very big restriction aspect clinical detection.As seen studying a kind of diagnostic reagent of fast detecting Avian pneumo-encephalitis virus so that epidemic situation is carried out scene monitoring in time, has been urgent problem during present herding is produced.
The utility model content
At the prior art above shortcomings, the purpose of this utility model provides a kind of simple in structure, easy to operate, quick, and responsive and the good Avian pneumo-encephalitis virus immune colloidal gold test card of specificity.
The purpose of this utility model is achieved in that a kind of ewcastle disease immune colloidal gold test card, is made of reagent strip and test cassette; Wherein, reagent strip is made up of backing, sample pad, pad, NC film and adsorptive pads, and in-plane is provided with the NC film on the backing, and the two ends of NC film are adjacent connection pad and adsorptive pads respectively; Sample pad is located at the top of pad; Be sprayed with golden labeling antibody Ab1 on the pad; Being sprayed with live width on the NC film is capture antibody Ab2 and the Quality Control antibody A b3 of 1mm; Pad is placed on the position that golden labeling antibody combines with NDV in the sample;
Test cassette is made up of box body and lid, and lid is provided with a well and a reading window, and well is over against sample pad; Read window over against the NC film; The middle part of box body is provided with the groove that is used for fixing described reagent strip, and the both sides of groove evenly are provided with and are used for the projection that is connected with lid.
Compared to existing technology, the utlity model has following advantage:
The utility model adopts the colloidal gold-labeled method principle, promptly uses collaurum as the trace labelling thing, according to the specificity of ewcastle disease antigen-antibody reaction; Physical characteristicss such as high electron density, grain size, shape and color reaction according to collaurum add the immuno-biology characteristic that bond has, and Avian pneumo-encephalitis virus hemagglutinin monoclonal antibody is carried out mark.Utilize the immune colloidal gold chromatography technology to make the colloid gold reagent bar again, this test strips can detect the Avian pneumo-encephalitis virus in ill animal blood, the ight soil.That this technology has is easy to operate, characteristics fast, and responsive and specificity is good.
The ewcastle disease immune colloidal gold test card is simple in structure, is easy to grasp use; And with low cost, be applicable to basic unit or clinical detection Avian pneumo-encephalitis virus; Can detect the Avian pneumo-encephalitis virus of bird blood, ight soil, go out the result about 10min, susceptibility reaches 10ng/ml, can be used for the quick diagnosis of ewcastle disease, is applicable to relate to plant, epidemic prevention quarantine departments and the medical institutions etc. that need ewcastle disease quick diagnosis or detection.
Description of drawings
Fig. 1 is the structural representation of the utility model reagent strip;
Fig. 2 is the structural representation of the utility model test cassette;
Fig. 3 is the structural representation of the utility model test card box body.
Embodiment
Below in conjunction with specific embodiment wooden utility model is described further.
As shown in Figure 1, a kind of ewcastle disease immune colloidal gold test card is made of reagent strip and test cassette; Wherein, reagent strip is made up of backing 1, sample pad 2, pad 3, cellulose nitrate NC film 4 and adsorptive pads 5; Backing 1, sample pad 2, pad 3, cellulose nitrate NC film 4 and adsorptive pads 5 are the suitable thin slice strip structure of width, the middle part on plane is provided with NC film 4 (being cellulose nitrate NC film) on the backing 1, and the two ends of cellulose nitrate NC film 4 are adjacent connection pad 3 and adsorptive pads 5 respectively; Sample pad 2 is located at the top of pad 3; Be sprayed with golden labeling antibody Ab1 (Avian pneumo-encephalitis virus monoclonal antibody) on the pad 3; Being sprayed with live width on the cellulose nitrate NC film 4 is capture antibody Ab2 (goat-anti NDV antibody) and the Quality Control antibody A b3 (the anti-Balb/C globulin antibody of rabbit) of 1mm; Pad 3 is placed on the position that golden labeling antibody combines with NDV in the sample.
As shown in Figures 2 and 3, test cassette fastens (clamping) by box body 11 and lid 8 and forms, and lid 8 is provided with a well 9 and a reading window 10, and well 9 is the positions that add sample over against sample pad 2; Read window 10 over against cellulose nitrate NC film 4, read window 10 sides and be printed on T and C letter, represent the position of NC film detection line and the position of nature controlling line respectively.The middle part of box body 11 is provided with the groove 12 that is used for fixing described reagent strip, and two outsides of groove 12 evenly are provided with and are used for the projection 13 that is connected with lid 8; Among the figure, the 14th, passive projection.
Use the method for the utility model fast detecting Avian pneumo-encephalitis virus, step is as follows:
1, the utility model utilizes the specificity and the simple principle fast of colloidal gold immunochromatographimethod of antigen-antibody reaction, set up Avian pneumo-encephalitis virus monoclonal antibody (Ab1, Antibody 1) and goat-anti Avian pneumo-encephalitis virus polyclonal antibody (Ab2, Antibody 2) and goat-anti Balb/C mouse immuning ball protein antibody (Ab3, Antibody 3);
2, determine pad optimization process liquid (80mM Na
2B
4O
710H
2O, 0.1% PVA, 0.1% Tween-20,0.3%BSA, described number percent is weight percent concentration), pad (glass fibre membrane) is soaked in the treating fluid under 4 ℃ of conditions, spend the night (about 12 hours); Oven dry (4-5h) under 37 ℃ of conditions then; Again with Ab1 mark collaurum and be sprayed on the pad; On cellulose nitrate NC film, be sprayed with capture antibody Ab2 that live width is 1mm (goat-anti NDV antibody) and Quality Control antibody A b3 (the anti-Balb/C globulin antibody of rabbit) then; Be assembled into reagent strip and test card respectively by pad, sample pad, backing, plastic clip etc. at last.
3, sample to be detected is added on the sample pad filter after, NDV in the sample combines with golden labeling antibody Ab1 in the pad, Ab2 first by the chromatography effect and on the NC film combines and forms visible joint line (detection line, be called for short the T line), not continuing chromatography in conjunction with intact golden labeling antibody combines with Ab3, form second joint line (nature controlling line is called for short the C line);
4, observations: two joint lines then appear in the positive; A C line then only appears in feminine gender; The invalid lines that then do not occur of reagent strip.
Above embodiment only is used to help those skilled in the art more comprehensively to understand the utility model, but the utility model is not imposed any restrictions.The replacement that all any this areas of making according to content of the present utility model are equal to all belongs within the scope of the utility model protection.
Claims (1)
1, a kind of ewcastle disease immune colloidal gold test card is made of reagent strip and test cassette; Wherein, reagent strip is made up of backing (1), sample pad (2), pad (3), NC film (4) and adsorptive pads (5), and backing (1) is gone up the middle part and is provided with NC film (4), and the two ends of NC film (4) are adjacent connection pad (3) and adsorptive pads (5) respectively; Sample pad (2) is located at the top of pad (3); Be sprayed with golden labeling antibody Ab1 on the pad (3); Being sprayed with live width on the NC film (4) is capture antibody Ab2 (6) and the Quality Control antibody A b3 (7) of 1mm; Pad (3) is placed on the position that golden labeling antibody combines with NDV in the sample;
Test cassette is made up of box body (11) and lid (8), and lid (8) is provided with a well (9) and a reading window (10), and well (9) is over against sample pad (2); Read window (10) over against NC film (4); The middle part of box body (11) is provided with the groove (12) that is used for fixing described reagent strip, and the both sides of groove (12) evenly are provided with and are used for the projection (13) that is connected with lid (8).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2008200984369U CN201203618Y (en) | 2008-05-29 | 2008-05-29 | Immune colloidal gold test card for new castle disease virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2008200984369U CN201203618Y (en) | 2008-05-29 | 2008-05-29 | Immune colloidal gold test card for new castle disease virus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN201203618Y true CN201203618Y (en) | 2009-03-04 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNU2008200984369U Expired - Fee Related CN201203618Y (en) | 2008-05-29 | 2008-05-29 | Immune colloidal gold test card for new castle disease virus |
Country Status (1)
| Country | Link |
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| CN (1) | CN201203618Y (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101819204A (en) * | 2010-05-28 | 2010-09-01 | 青岛康地恩药业有限公司 | Collaurum test paper as well as preparation method and application thereof |
| CN101852802A (en) * | 2010-05-05 | 2010-10-06 | 青岛康地恩药业有限公司 | Test paper for detecting newcastle disease and infectious bronchitis viruses in one step |
| CN102608331A (en) * | 2012-03-16 | 2012-07-25 | 吉林大学 | Colloidal gold labeled quick-diagnosis test strip for virulent strains and low-virulent strains of Newcastle disease |
| CN103140758A (en) * | 2010-09-24 | 2013-06-05 | 基立福疗法公司 | Immunochromatography devices, methods and kits |
| CN104459142A (en) * | 2014-12-12 | 2015-03-25 | 河南省农业科学院 | Test paper for distinguishing virulent strain from attenuated strain of newcastle disease virus |
-
2008
- 2008-05-29 CN CNU2008200984369U patent/CN201203618Y/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101852802A (en) * | 2010-05-05 | 2010-10-06 | 青岛康地恩药业有限公司 | Test paper for detecting newcastle disease and infectious bronchitis viruses in one step |
| CN101819204A (en) * | 2010-05-28 | 2010-09-01 | 青岛康地恩药业有限公司 | Collaurum test paper as well as preparation method and application thereof |
| CN103140758A (en) * | 2010-09-24 | 2013-06-05 | 基立福疗法公司 | Immunochromatography devices, methods and kits |
| CN102608331A (en) * | 2012-03-16 | 2012-07-25 | 吉林大学 | Colloidal gold labeled quick-diagnosis test strip for virulent strains and low-virulent strains of Newcastle disease |
| CN104459142A (en) * | 2014-12-12 | 2015-03-25 | 河南省农业科学院 | Test paper for distinguishing virulent strain from attenuated strain of newcastle disease virus |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090304 Termination date: 20110529 |