CN106443001A - Avian influenza detection kit - Google Patents
Avian influenza detection kit Download PDFInfo
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- CN106443001A CN106443001A CN201611072118.0A CN201611072118A CN106443001A CN 106443001 A CN106443001 A CN 106443001A CN 201611072118 A CN201611072118 A CN 201611072118A CN 106443001 A CN106443001 A CN 106443001A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
- G01N33/561—Immunoelectrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/11—Orthomyxoviridae, e.g. influenza virus
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Abstract
The invention discloses an avian influenza detection kit, comprising: a chromatographic medium fixedly provided with a first antibody having antigen-antibody reaction with H5 subtype avian influenza virus nucleoprotein but substantially having no antigen-antibody reaction with H7 subtype avian influenza virus nucleoprotein; and a marking agent formed by combining a marker to a second antibody having antigen-antibody reaction with H5 subtype avian influenza virus nucleoprotein but substantially having no antigen-antibody reaction with H7 subtype avian influenza virus nucleoprotein; as the avian influenza detection kit has high detection sensitivity, 'positive' determination can be made with less viral quantity than existing kits, false-negative determinations are fewer, and 'negative' determination reliability is very high. Therefore, for a patient in initial indigence of avian influenza and shortly after the virus starts proliferates in the body, beneficial information necessary for a doctor to make correct diagnosis on influenza infections can be provided, and treatment with anti-influenza virus agents can be provided in early stage.
Description
Technical field
The present invention relates to a kind of detection method, specifically a kind of detection kit of avian influenza.
Background technology
Avian influenza is a kind of acute, the deadly infectious disease of the birdss being caused by orthomyxovirus section influenza A, except
Also can infect the many animals such as people, pig and horse, A class infectious disease is classified as by OIE outside infected poultry.Bird flu viruss (AIV)
Belong to influenza A, frequently, blood serum subtype is numerous for bird flu viruss (AIV) variation.It is divided into 16 according to HA antigenic specificity
Individual HA hypotype.Antigenic variability is strong, and extensively, between hypotype, no intersecting protective, so that bird flu frequently breaks out, becomes host range
One War Torn disease of aviculture.
How not high pathogenic avian influenza is mainly caused by H5 or H7 hypotype AIV, and incubation period is extremely short, break out suddenly,
See any clinical symptoms and sudden death, M ＆ M may be up to 100%, brings serious financial consequences to aviculture,
In addition also can infect people, even result in death.According to World Health Organization (WHO) (WHO), by the end of May 22 in 2009
Day, the whole world total H5N1 subtype avian influenza virus infection people 429, dead 262；China has cases of infection
38, dead 25.At present, middle low pathogenicity bird flu based on H5 hypotype for China some areas is popular, can cause chicken
Egg production declines, and causes very big economic loss to China's aviculture, this point has caused the extensive concern of domestic all circles.So
Early stage quick detection is undoubtedly prevention, controls the precondition of bird flu, in particular for the AIV of H5, H7 hypotype.Virus point
So far it is still diagnosis AI from, serological test and commonly used method of shaping is carried out to AIV.But these method complex operations
Complicated；It is difficult to carry out quick diagnosis to AI；Extremely it is unfavorable for the preventing and treating of AI.
Content of the invention
It is an object of the invention to provide a kind of detection kit of avian influenza, to solve to propose in above-mentioned background technology
Problem.
For achieving the above object, the present invention provides following technical scheme：
A kind of detection kit of avian influenza, contains：It is fixed with and have antigen antibody reaction with H5 type bird flu viruss nucleoprotein
And with H7 type bird flu viruss nucleoprotein essentially without the chromatography media of the first antibody of antigen antibody reaction and with H5 type
Bird flu viruss nucleoprotein has antigen antibody reaction and with H7 type bird flu viruss nucleoprotein essentially without antigen antibody reaction
Second antibody and the labelled reagent that is combined into of label.
As the further scheme of the present invention：Described first antibody is by being divided with by SDS- polyacrylamide gel electrophoresis
From the H5 type bird flu viruss nucleoprotein of total length there is no the antibody of antigen antibody reaction in Western blot method, second
Antibody is to exist with by the H5 type bird flu viruss nucleoprotein of the separated total length of SDS- polyacrylamide gel electrophoresis
There is the antibody of antigen antibody reaction in Western blot method.
As the further scheme of the present invention：Described first antibody is the monoclonal antibody of a kind or more than a kind.
As the further scheme of the present invention：Described first antibody is by the total length to influenza A H5N1 hypotype
Nucleoprotein carry out antibody obtained from immunity.
As the further scheme of the present invention：A kind of avian influenza detection method, it is using immunochromatographic method
Detection method, employ be fixed with have antigen antibody reaction with H5 type bird flu viruss nucleoprotein and with H7 type bird flu viruss core
Albumen has essentially without the chromatography media of the first antibody of antigen antibody reaction and with H5 type bird flu viruss nucleoprotein
Antigen antibody reaction and with H7 type bird flu viruss nucleoprotein essentially without the second antibody of antigen antibody reaction and label
The labelled reagent being combined into, wherein, first antibody is and the H5 by the separated total length of SDS- polyacrylamide gel electrophoresis
Type bird flu viruss nucleoprotein does not have the antibody of antigen antibody reaction in Western blot method, second antibody be with by
The H5 type bird flu viruss nucleoprotein of the separated total length of SDS- polyacrylamide gel electrophoresis is in Western blot method
There is the antibody of antigen antibody reaction.
As the further scheme of the present invention：Described first antibody is the monoclonal antibody of a kind or more than a kind.
As the further scheme of the present invention：Described first antibody is by the total length to influenza A H5N1 hypotype
Nucleoprotein carry out antibody obtained from immunity.
Compared with prior art, the invention has the beneficial effects as follows：The avian influenza virus detection kit of the present invention due to
Detection sensitivity is high, and therefore few than existing detection kit virus quantity just can obtain the judgement of " positive ", therefore, false cloudy
Property judgement reduce, very high is become to the reliability of the judgement of " negative ".Therefore, for being in avian influenza their early stage
And virus starts to breed patient soon in vivo, using the teaching of the invention it is possible to provide doctor makes having needed for the correct diagnosis of influenza infection
Beneficial information, and the treatment with Anti-influenza virus agent can be started in early stage.Even if in addition, suppressing disease using vaccination
It is also possible to correctly diagnosis has or not the infection of avian influenza virus, therefore, it is possible to cause to infection in the patient of malicious growth rate
The attention expanding, in addition it is also possible to provide for judging important information in the immunology of avian influenza vaccine effect.
Specific embodiment
With reference to specific embodiment, the technical scheme of this patent is described in more detail.
A kind of detection kit of avian influenza, contains：It is fixed with and have antigen-antibody with H5 type bird flu viruss nucleoprotein
Reaction and with H7 type bird flu viruss nucleoprotein essentially without the chromatography media of the first antibody of antigen antibody reaction, Yi Jiyu
H5 type bird flu viruss nucleoprotein has antigen antibody reaction and with H7 type bird flu viruss nucleoprotein essentially without antigen-antibody
The second antibody of reaction and the labelled reagent that is combined into of label.
First antibody is and the H5 type bird flu viruss core by the separated total length of SDS- polyacrylamide gel electrophoresis
Albumen does not have the antibody of antigen antibody reaction in Western blot method, and second antibody is and by SDS- polyacrylamide
The H5 type bird flu viruss nucleoprotein of the separated total length of gel electrophoresiss has antigen antibody reaction in Western blot method
Antibody.
First antibody is the monoclonal antibody of a kind or more than a kind.
First antibody is to carry out resisting obtained from immunity by the nucleoprotein of the total length to influenza A H5N1 hypotype
Body.
A kind of avian influenza detection method, it is the detection method using immunochromatographic method, employ be fixed with
H5 type bird flu viruss nucleoprotein has antigen antibody reaction and with H7 type bird flu viruss nucleoprotein essentially without antigen-antibody
The chromatography media of first antibody of reaction and have an antigen antibody reaction with H5 type bird flu viruss nucleoprotein and with H7 type fowl
Influenza nucleoprotein essentially without the labelled reagent that is combined into of second antibody and label of antigen antibody reaction, its
In, first antibody is to exist with by the H5 type bird flu viruss nucleoprotein of the separated total length of SDS- polyacrylamide gel electrophoresis
There is no the antibody of antigen antibody reaction, second antibody is and by SDS- polyacrylamide gel electrophoresis in Western blot method
The H5 type bird flu viruss nucleoprotein of separated total length has the antibody of antigen antibody reaction in Western blot method.
First antibody is the monoclonal antibody of a kind or more than a kind.
First antibody is to carry out resisting obtained from immunity by the nucleoprotein of the total length to influenza A H5N1 hypotype
Body.
Used in the present invention, first antibody and second antibody are to have antigen-antibody anti-with H5 type bird flu viruss nucleoprotein
Ying Eryu B type influenza nucleoprotein is essentially without the antibody of antigen antibody reaction.Influenza virus are anti-due to nucleoprotein
The difference of originality and be divided into A type, B type etc..And then, H5 type bird flu viruss have hemagglutinin on the surface of virion
(HA) it is divided into and the such glycoprotein of neuraminidase (NA), and due to the architectural difference of these HA and NA each
Plant hypotype.First antibody used in the present invention and second antibody, therefore can be with due to identifying the nucleoprotein of influenza virus
Widely there is antigen antibody reaction in the nucleoprotein of the various hypotypes of H5 type bird flu viruss, at least can be with avian influenza disease
There is antigen antibody reaction in the nucleoprotein of the H1N1 hypotype of poison, H3N2 hypotype, H5N1 hypotype and H7N7 hypotype etc., but
It is the antibody not having antigen antibody reaction with the nucleoprotein of B type influenza virus.Anti- with the first antibody of the present invention and second
It can be detached native protein from virus that body occurs the nucleoprotein of the H5 type bird flu viruss of antigen antibody reaction, also may be used
To be the recombiant protein that the base sequence based on known nucleoprotein gene makes.And then, the first antibody and with the present invention
Two antibody occur antigen antibody reactions nucleoprotein can be isolate and purify from viral constituent or not
Purification, but it is also possible to be derived from exposed to contact with nucleoprotein and antibody with surfactant in the case of unsegregated
The nucleoprotein of the virus that mode was processed.
" unmodified H5 type bird flu viruss nucleoprotein " in the present invention, as long as remain naturally occurring H5 type fowl stream
There is the sufficient stereochemical structure of antigen antibody reaction in the stereochemical structure of susceptible toxalbumin, at least maintenance and specific antibody
But, because SDS-PAGE etc. destroys the stereochemical structure of this albumen naturally occurring, is unable to maintain that H5 type avian influenza
Except malicious nucleoprotein is to the substantial antigen antibody reaction of this antibody.
Whether the first antibody used in the present invention and second antibody occur antigen-antibody anti-with influenza nucleoprotein
Should be confirmed using known method of immunity.That is, if to classify with determination form, there is sandwich, competing
Strive the method for immunity such as method, coacervation, if with using label classify, have the immunity such as fluorescence method, enzyme process, radioactive method
Assay method, all can carry out the confirmation of antigen antibody reaction using any one in these method of immunity.At this
In bright, " essentially without antigen antibody reaction " refers to, in above-mentioned method of immunity, not anti-in detectable level
Even if antigen-antibody reaction or the journey having the degree reacting this reaction and the antigen antibody reaction of H5 type bird flu viruss nucleoprotein
Degree compares reaction that is substantially weaker, being same degree with the other albumen constituting influenza virus, is not specific reaction.
As the first antibody of the present invention, be to have antigen antibody reaction with influenza A nucleoprotein and with B type influenza
Virus nucleoprotein essentially without the antibody of antigen antibody reaction, and then, as long as with naturally occurring A type influenza virus core
This albumen that the stereochemical structure at the position of generation antigen antibody reaction of albumen has been destroyed is essentially without antigen antibody reaction
Antibody, as such antibody, for example, it is preferable to be to have antigen antibody reaction and and B with influenza A nucleoprotein
Type influenza nucleoprotein essentially without the antibody of antigen antibody reaction, with the A type influenza by the detached total length of SDS-PAGE
Virus nucleoprotein does not have the antibody of antigen antibody reaction in Western blot method.
Claims (7)
1. a kind of detection kit of avian influenza is it is characterised in that contain：It is fixed with and have with H5 type bird flu viruss nucleoprotein
Antigen antibody reaction and with H7 type bird flu viruss nucleoprotein essentially without the first antibody of antigen antibody reaction chromatography be situated between
Matter and have antigen antibody reaction with H5 type bird flu viruss nucleoprotein and with H7 type bird flu viruss nucleoprotein essentially without
The labelled reagent that the second antibody of antigen antibody reaction and label are combined into.
2. a kind of detection kit of avian influenza according to claim 1 is it is characterised in that described first antibody is
With the H5 type bird flu viruss nucleoprotein by the separated total length of SDS- polyacrylamide gel electrophoresis in Western blot
There is no the antibody of antigen antibody reaction in method, second antibody be with by the separated total length of SDS- polyacrylamide gel electrophoresis
H5 type bird flu viruss nucleoprotein have the antibody of antigen antibody reaction in Western blot method.
3. a kind of detection kit of avian influenza according to claim 1 is it is characterised in that described first antibody is 1
Kind or more than a kind of monoclonal antibody.
4. a kind of detection kit of the avian influenza according to claim 1 or 3 is it is characterised in that described first antibody
It is that antibody obtained from immunity is carried out by the nucleoprotein of the total length to influenza A H5N1 hypotype.
5. a kind of avian influenza detection method, it is the detection method using immunochromatographic method, employs and is fixed with and H5
Type bird flu viruss nucleoprotein has antigen antibody reaction and anti-essentially without antigen-antibody with H7 type bird flu viruss nucleoprotein
The chromatography media of the first antibody answered and have an antigen antibody reaction with H5 type bird flu viruss nucleoprotein and with H7 type fowl stream
Influenza Virus nucleoprotein essentially without the labelled reagent that is combined into of second antibody and label of antigen antibody reaction, wherein,
First antibody is to exist with by the H5 type bird flu viruss nucleoprotein of the separated total length of SDS- polyacrylamide gel electrophoresis
There is no the antibody of antigen antibody reaction, second antibody is and by SDS- polyacrylamide gel electrophoresis in Western blot method
The H5 type bird flu viruss nucleoprotein of separated total length has the antibody of antigen antibody reaction in Western blot method.
6. a kind of detection kit of avian influenza according to claim 4 is it is characterised in that described first antibody is 1
Kind or more than a kind of monoclonal antibody.
7. a kind of detection kit of avian influenza according to claim 4 is it is characterised in that described first antibody is logical
The nucleoprotein crossing the total length to influenza A H5N1 hypotype carries out antibody obtained from immunity.
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| CN201611072118.0A CN106443001A (en) | 2016-11-29 | 2016-11-29 | Avian influenza detection kit |
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| CN201611072118.0A CN106443001A (en) | 2016-11-29 | 2016-11-29 | Avian influenza detection kit |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1591014A (en) * | 2003-09-03 | 2005-03-09 | 北京阿斯可来生物工程有限公司 | Influenza Virus A colloidal gold quick detection test paper |
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| CN101591390A (en) * | 2008-05-30 | 2009-12-02 | 中国科学院上海生命科学研究院 | The monoclonal antibody and the application thereof of the avian influenza virus NP in anti-H5N1 source |
| CN201397335Y (en) * | 2009-05-18 | 2010-02-03 | 中国农业科学院哈尔滨兽医研究所 | A test paper strip for Colloidal gold test for detecting H5N1 subtype avian influenza virus rapidly |
| JP2012112879A (en) * | 2010-11-26 | 2012-06-14 | National Center For Global Health & Medicine | Immunity examination reagent, and examination device and method using the same |
| CN104246504A (en) * | 2012-03-30 | 2014-12-24 | 田中贵金属工业株式会社 | Detection kit for influenza a virus |
-
2016
- 2016-11-29 CN CN201611072118.0A patent/CN106443001A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1591014A (en) * | 2003-09-03 | 2005-03-09 | 北京阿斯可来生物工程有限公司 | Influenza Virus A colloidal gold quick detection test paper |
| CN101553732A (en) * | 2006-08-31 | 2009-10-07 | 大阪府 | Rapid diagnosis method specific to avian influenza virus |
| CN101591390A (en) * | 2008-05-30 | 2009-12-02 | 中国科学院上海生命科学研究院 | The monoclonal antibody and the application thereof of the avian influenza virus NP in anti-H5N1 source |
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| JP2012112879A (en) * | 2010-11-26 | 2012-06-14 | National Center For Global Health & Medicine | Immunity examination reagent, and examination device and method using the same |
| CN104246504A (en) * | 2012-03-30 | 2014-12-24 | 田中贵金属工业株式会社 | Detection kit for influenza a virus |
Non-Patent Citations (1)
| Title |
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| 孙恩成: "基因免疫制备抗甲型禽流感病毒核蛋白特异性单克隆抗体", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
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Application publication date: 20170222 |