CN107271677A - A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus - Google Patents
A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus Download PDFInfo
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- CN107271677A CN107271677A CN201710684488.8A CN201710684488A CN107271677A CN 107271677 A CN107271677 A CN 107271677A CN 201710684488 A CN201710684488 A CN 201710684488A CN 107271677 A CN107271677 A CN 107271677A
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- epidemic diarrhea
- diarrhea virus
- porcine epidemic
- immunity biosensor
- monoclonal antibody
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- 241001135549 Porcine epidemic diarrhea virus Species 0.000 title claims abstract description 28
- 230000036039 immunity Effects 0.000 title claims abstract description 18
- 239000011159 matrix material Substances 0.000 claims abstract description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229910021389 graphene Inorganic materials 0.000 claims abstract description 6
- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000001556 precipitation Methods 0.000 claims abstract description 6
- 229960003351 prussian blue Drugs 0.000 claims abstract description 6
- 239000013225 prussian blue Substances 0.000 claims abstract description 6
- 206010012735 Diarrhoea Diseases 0.000 claims description 14
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 8
- 229940098773 bovine serum albumin Drugs 0.000 claims description 8
- 239000012888 bovine serum Substances 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 10
- 241000282898 Sus scrofa Species 0.000 description 14
- 241000700605 Viruses Species 0.000 description 9
- 238000000034 method Methods 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 201000010099 disease Diseases 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 208000005156 Dehydration Diseases 0.000 description 1
- 208000005577 Gastroenteritis Diseases 0.000 description 1
- 206010022678 Intestinal infections Diseases 0.000 description 1
- 241000702670 Rotavirus Species 0.000 description 1
- 229910021607 Silver chloride Inorganic materials 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000002484 cyclic voltammetry Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 231100000517 death Toxicity 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 230000005226 mechanical processes and functions Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
Classifications
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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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pathology (AREA)
- Hematology (AREA)
- General Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Electrochemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus, it includes being located at graphene modified layer, the Prussian blue beds of precipitation and Porcine epidemic diarrhea virus monoclonal antibody layer that electrode matrix surface is stacked gradually from bottom to top.The present invention is combined by graphene modified layer, the Prussian blue beds of precipitation and Porcine epidemic diarrhea virus monoclonal antibody layer with electrode matrix, form immunity biosensor, Porcine epidemic diarrhea virus is determined so as to quick being set suitable for non-laboratory, it is easy to the Site Detection of clinical quick detection, such as plant of basic unit.
Description
Technical field
The present invention relates to field of biosensors, more particularly to a kind of immune life for being used to detect pig fluidity diarrhea virus
Thing sensor.
Background technology
Pig epidemic diarrhea (porcine epidemic diarrhea, PED) is a kind of high degree in contact enteric infection
Disease, clinically causes grice diarrhoea, vomiting, dehydration and the high fatal rate of suckling pig, is that many pig-raising countries in the current world are young
One of important epidemic disease of pig Deaths, huge economic loss is brought to global pig industry.The disease is in clinical symptoms
Upper is difficult to differentiate with the epidemic disease such as transmissible gastroenteritis of swine, rotavirus illness and colibacillosis, is easily caused mistaken diagnosis and prevents and treats not
When.At present, the sick diagnostic method mainly has：Virus Isolation, immunofluorescence technique (IF), immunohistochemistry technology (IHT),
Immunoelectron microscopic method (IEM), serum neutralization test, ELISA and RT-PCR method etc..At present using more for ELISA method and PCR
Method, both approaches are so that its is practical, efficient, quick, accurate and is widely used.But, although two methods can be right
Pig prevalence diarrhea virus carries out qualitative and quantitative analysis, but due to being limited by instrument and equipment, it is time-consuming long, while detection device
Be inconvenient to carry, inconvenient is still suffered from plant of basic unit and Site Detection.How the showing of Porcine epidemic diarrhea virus is made
Field detection has feasibility, is a problem of current industry.
The content of the invention
The technical problem to be solved in the present invention is：It is used to detect that pig is flowed there is provided one kind for above-mentioned the deficiencies in the prior art
The immunity biosensor of row diarrhea virus, it causes the Site Detection of pig epidemic diarrhea to have feasibility.
The present invention solve its technical problem solution be：A kind of immune life for being used to detect Porcine epidemic diarrhea virus
Thing sensor, it includes being located at graphene modified layer, the Prussian blue beds of precipitation that electrode matrix surface is stacked gradually from bottom to top
With Porcine epidemic diarrhea virus monoclonal antibody layer.
As the further improvement of above-mentioned technical proposal, the Porcine epidemic diarrhea virus monoclonal antibody layer is by concentration
50 μ g/mL Porcine epidemic diarrhea virus monoclonal antibody solution is dried and formed.
As the further improvement of above-mentioned technical proposal, the immunity biosensor also includes being located at the pig epidemic
Bovine serum albumin confining bed in diarrhea virus monoclonal antibody layer surface.Further, the bovine serum albumin confining bed is by dense
The bovine serum albumen solution spent for 1~2%, which is dried, to be formed.
As the further improvement of above-mentioned technical proposal, the electrode matrix is glass-carbon electrode.
The beneficial effects of the invention are as follows：The present invention passes through graphene modified layer, the Prussian blue beds of precipitation and pig epidemic
Diarrhea virus monoclonal antibody layer is combined with electrode matrix, forms immunity biosensor, so as to be set suitable for non-laboratory
In it is quick determine Porcine epidemic diarrhea virus, be easy to the Site Detection of clinical quick detection, such as plant of basic unit.
Brief description of the drawings
Technical scheme in order to illustrate the embodiments of the present invention more clearly, makes required in being described below to embodiment
Accompanying drawing is briefly described.Obviously, described accompanying drawing is a part of embodiment of the present invention, rather than is all implemented
Example, those skilled in the art on the premise of not paying creative work, can also obtain other designs according to these accompanying drawings
Scheme and accompanying drawing.
Fig. 1 is the Rotating fields schematic diagram of the present invention.
Embodiment
The technique effect of the design of the present invention, concrete structure and generation is carried out below with reference to embodiment and accompanying drawing clear
Chu, complete description, to be completely understood by the purpose of the present invention, feature and effect.Obviously, described embodiment is this hair
Bright a part of embodiment, rather than whole embodiments, based on embodiments of the invention, those skilled in the art is not paying
The other embodiment obtained on the premise of creative work, belongs to the scope of protection of the invention.In addition, be previously mentioned in text
All connection/annexations, not singly refer to component and directly connect, and refer to be added deduct by adding according to specific implementation situation
Few couple auxiliary, to constitute more excellent draw bail.Each technical characteristic in the invention, in not conflicting conflict
Under the premise of can be with combination of interactions.
Reference picture 1, a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus, it includes being located at electrode base
Graphene modified layer 2, the Prussian blue beds of precipitation 3 and Porcine epidemic diarrhea virus Dan Ke that the surface of body 1 is stacked gradually from bottom to top
Grand antibody layer 4.
It is further used as preferred embodiment, the Porcine epidemic diarrhea virus monoclonal antibody layer 4 is 50 μ by concentration
G/mL Porcine epidemic diarrhea virus monoclonal antibody viral solution is dried and formed.Specifically, under the concentration conditions pig is popular
Diarrhea virus monoclonal antibody solution has good stability, and it can make the popular diarrhea virus monoclonal antibody layer of the pig to be formed
4 have excellent surface physicochemical property.
It is further used as preferred embodiment, the immunity biosensor also includes being located at the Porcine Epidemic Diarrhea
Bovine serum albumin confining bed 5 on malicious 4 surface of monoclonal antibody layer, the bovine serum albumin confining bed 5 can play signal amplification
Effect so that the immunity biosensor has good sensitivity and the degree of accuracy.Further, the bovine serum albumin closing
Layer 5 is formed by concentration for 1~2% bovine serum albumen solution drying.Specifically, the bovine serum albumen solution under the concentration conditions
With good stability, it can make the bovine serum albumin confining bed 5 to be formed have excellent surface physicochemical property.
It is further used as preferred embodiment, the electrode matrix 1 is glass-carbon electrode.
The present invention is applied to the Site Detection of Porcine epidemic diarrhea virus.In actual mechanical process, first by the present invention's
Immunity biosensor immerses 30min in 100 μ L Porcine epidemic diarrhea virus (15 μ g/mL), and taking-up is eluted with double distilled water, so
After immerse 30min in 50 μ g/mL Porcine epidemic diarrhea virus polyclonal antibodies (50 μ g/mL), taking-up is eluted with double distilled water, with
Working electrode is dipped into 50~200 μ L testing samples and is immunized as working electrode by this immunity biosensor through processing
30min~120min is reacted, the immunity biosensor PBS is taken out after capture is viral and cleans, with Ag/AgCl electrodes
For reference electrode, platinum electrode detects the Porcine epidemic diarrhea virus in sample, to complete pig as to electrode with cyclic voltammetry
The Site Detection of epidemic diarrhea virus.
The better embodiment to the present invention is illustrated above, but the invention is not limited to the implementation
Example, those skilled in the art can also make a variety of equivalent modifications or replace on the premise of without prejudice to spirit of the invention
Change, these equivalent modifications or replacement are all contained in the application claim limited range.
Claims (5)
1. a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus, it is characterised in that：Including positioned at electrode base
Graphene modified layer (2), the Prussian blue beds of precipitation (3) and Porcine Epidemic Diarrhea that body (1) surface is stacked gradually from bottom to top
Malicious monoclonal antibody layer (4).
2. a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus according to claim 1, its feature
It is：The Porcine epidemic diarrhea virus monoclonal antibody layer (4) is by Porcine epidemic diarrhea virus Dan Ke of the concentration for 50 μ g/mL
Grand antibody-solutions are dried and formed.
3. a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus according to claim 1, its feature
It is：Also include the bovine serum albumin confining bed being located on Porcine epidemic diarrhea virus monoclonal antibody layer (4) surface
(5)。
4. a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus according to claim 3, its feature
It is：The bovine serum albumin confining bed (5) is formed by concentration for 1~2% bovine serum albumen solution drying.
5. a kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus according to claim 1, its feature
It is：The electrode matrix (1) is glass-carbon electrode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710684488.8A CN107271677A (en) | 2017-08-11 | 2017-08-11 | A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710684488.8A CN107271677A (en) | 2017-08-11 | 2017-08-11 | A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107271677A true CN107271677A (en) | 2017-10-20 |
Family
ID=60080298
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710684488.8A Pending CN107271677A (en) | 2017-08-11 | 2017-08-11 | A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus |
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| Country | Link |
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| CN (1) | CN107271677A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112986356A (en) * | 2019-12-12 | 2021-06-18 | 福建海峡石墨烯产业技术研究院有限公司 | Graphene field effect transistor biosensor and preparation method thereof |
| CN115656304A (en) * | 2022-12-13 | 2023-01-31 | 佛山科学技术学院 | Immune biosensor for detecting PEDV and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101339192A (en) * | 2008-08-28 | 2009-01-07 | 河南省农业科学院 | Test paper for one-step detection for pig virus diarrhoea disease pathogen |
| US20130334042A1 (en) * | 2010-12-06 | 2013-12-19 | Syngenta Limited | Pathogen sensor |
| CN103777010A (en) * | 2014-01-27 | 2014-05-07 | 深圳市宝舜泰生物医药股份有限公司 | ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for porcine epidemic diarrhea viruses and preparation method thereof |
| CN207036860U (en) * | 2017-08-11 | 2018-02-23 | 佛山科学技术学院 | A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus |
-
2017
- 2017-08-11 CN CN201710684488.8A patent/CN107271677A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101339192A (en) * | 2008-08-28 | 2009-01-07 | 河南省农业科学院 | Test paper for one-step detection for pig virus diarrhoea disease pathogen |
| US20130334042A1 (en) * | 2010-12-06 | 2013-12-19 | Syngenta Limited | Pathogen sensor |
| CN103777010A (en) * | 2014-01-27 | 2014-05-07 | 深圳市宝舜泰生物医药股份有限公司 | ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for porcine epidemic diarrhea viruses and preparation method thereof |
| CN207036860U (en) * | 2017-08-11 | 2018-02-23 | 佛山科学技术学院 | A kind of immunity biosensor for being used to detect Porcine epidemic diarrhea virus |
Non-Patent Citations (4)
| Title |
|---|
| DEXIANG FENG ET AL: "Label-free electrochemical immunosensor for the carcinoembryonic antigen using a glassy carbon electrode modified with electrodeposited Prussian Blue, a graphene and carbon nanotube assembly and an antibody immobilized on gold nanoparticles", 《MICROCHIM ACTA》 * |
| 于鲁平等: "基于石墨烯-依来铬青蓝R的新型免标记电化学免疫传感器用于CA19-9检测", 《福建医科大学学报》 * |
| 张洁等: "采用石墨烯修饰的电化学免疫传感器检测猪肉中的己二烯雌酚含量", 《中国药科大学学报》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112986356A (en) * | 2019-12-12 | 2021-06-18 | 福建海峡石墨烯产业技术研究院有限公司 | Graphene field effect transistor biosensor and preparation method thereof |
| CN115656304A (en) * | 2022-12-13 | 2023-01-31 | 佛山科学技术学院 | Immune biosensor for detecting PEDV and preparation method and application thereof |
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| PB01 | Publication | ||
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| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Li Zhili Inventor after: Zhao Yunxiang Inventor after: Huang Shujian Inventor after: Liao Jiedan Inventor after: Huang Xingguo Inventor after: Qin Li Mei Inventor after: Liu Youming Inventor after: Chi Shihong Inventor before: Li Zhili Inventor before: Zhao Yunxiang Inventor before: Huang Shujian Inventor before: Liao Jiedan Inventor before: Huang Xingguo Inventor before: Qin Li Mei Inventor before: Liu Youming Inventor before: Chi Shi Hong |
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Application publication date: 20171020 |
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