CN109613272A - A kind of protein chip and preparation method thereof for the detection of pork veterinary drug residue - Google Patents
A kind of protein chip and preparation method thereof for the detection of pork veterinary drug residue Download PDFInfo
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- CN109613272A CN109613272A CN201910022566.7A CN201910022566A CN109613272A CN 109613272 A CN109613272 A CN 109613272A CN 201910022566 A CN201910022566 A CN 201910022566A CN 109613272 A CN109613272 A CN 109613272A
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
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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
- G01N33/531—Production of immunochemical test materials
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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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
- G01N33/552—Glass or silica
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Abstract
The invention discloses a kind of for detecting the protein chip of six kinds of residues of veterinary drug in pork, and the preparation method of the protein chip includes the following steps: (1) black slide pretreatment；(2) antigenic solution point sample；(3) closing process；The protein chip is made.Protein chip of the present invention realizes that joint-detection can effectively improve detection efficiency for detecting pork veterinary drug residue, using the protein chip technology of our company's original creation, reduces testing cost；Inventor developed simultaneously include six indexs of CLEN, RAC, SAL, CAP, CPZ and AMC protein chip diagnostic kit, have quickly, efficiently, it is inexpensive the advantages that.
Description
Technical field
The present invention relates to field of biotechnology, more particularly, to a kind of for detecting the protein chip and its system of residue of veterinary drug
Preparation Method.
Background technique
Interior livestock and poultry body or product mesarcs drug or its metabolite are accumulated or retained in residue of veterinary drug after referring to medication.Beast
Correct, the reasonable use of medicine brings high benefit to aquaculture, but always has groups of people to pursue economic interests simply, produces in livestock and poultry
It illegally uses, abuse in industry links, add in violation of rules and regulations, injecting veterinary drug or harmful substance.The residual and harmful substance of veterinary drug are tight
Ghost image rings the sustainable development of animal food safety, animal husbandry, is more seriously directly detrimental to health and life security,
It is very big to human health risk.Therefore, it is necessary to carry out residue of veterinary drug content and forbidden drug and harmful substance to animal food
Detection, the problem livestock products of preventing come into the market each link, guarantee " the dining table safety " of people.
The index of six detection of veterinary drugs in food is respectively as follows: CLEN, RAC, SAL, CAP, CPZ, AMC.
It is a kind of adrenal neurostimulant that CLEN (clenobuterol hydrochloride), which is also known as " clenbuterol hydrochloride ", can change animal body
Interior metabolic pathway promotes the synthesis of protein in muscle, especially skeletal muscle, inhibits the synthesis of fat, to accelerate to grow
Speed.The meat containing clenobuterol hydrochloride residue has been eaten for a long time in people, can cause elevation of the blood pressure, blood vessel dilatation, palpitating speed,
The symptoms such as aggravation, neuroticism, Nausea and vomiting are breathed, more seriously it can induce and aggravate the state of an illness of arrhythmia cordis patient.
RAC (Ractopamine) is a kind of β agonist, is one kind of clenbuterol hydrochloride, as animal feed additive, to help
Long pig, ox, turkey bear muscle, reduce body fat.Pork containing Ractopamine once passes through food chain and enters human body, meeting
Greatly harm is generated to health, the sign of poisoning may be caused, nausea, dizziness, muscle tremors, palpitaition, blood pressure rise, promote
Cardiovascular disease influences reproductive system etc..
SAL (salbutamol) is a kind of short-acting beta 2-adrenergic receptor agonists, is used as antiasthmatic.Add salbutamol
In animal feeding-stuff, the cutability and meat change rate, reduction fat of livestock can be increased.It is easy that the food containing salbutamol is eaten for a long time
Appearance trembles, Nausea and vomiting, continues the symptoms such as increased heart rate or abnormal strong, the mood dysphoria of heartbeat.
CAP (chloramphenicol) can be used for the 50s subunit of bacterium ribosome, the synthesis of protein be obstructed, to gram
Positive, negative bacterium has inhibiting effect, the especially effect to typhoid bacillus, Bacillus influenzae, para-influenza Bacillus and Bordetella pertussis
It is better than other antibiotic.Chloramphenicol can inhibit marrow hemopoiesis function, cause granulocyte and decrease of platelet, aplastic poor
Blood, long with can also cause optic neuritis, incoordination and suprainfection.
CPZ (chlorpromazine) also known as wintermin are a kind of sleeping, calmness, hypnotic drug, are mainly used as in veterinary clinic
Sedative.Chlorpromazine is added in feed, other than achieving the effect that calm, hypnosis, moreover it is possible to play fattening indirectly and promote height work
With.Food of the long-term consumption containing chlorpromazine can cause oligoleukocythemia, dry, it is out of strength, drowsiness, constipation, palpitaition, under blood pressure
Drop, even shock etc..
AMC (Amoxicillin) is one kind in beta-lactam antibiotic, and bactericidal effect is strong, the ability of penetrating cell wall
By force, there is powerful antibacterial and bactericidal effect to the pathogenic G+ bacterium of majority and G- bacterium (including coccus and bacillus), widely used
In the infection for the urethra, respiratory tract and skin for treating various animals.Long-term accumulation may occur in which allergic symptom, digestion in human body
Systemic symptom, hematoligical symptom, mucocutaneous reaction, hepatic and renal function disorder, central nervous system symptom.
This six indexs are detected by extensive uses such as enterprise, testing agency, law enforcement agencies, government at present.But at present
The main stream approach taken is the measuring method (such as ELISA and colloidal gold etc.) of immune single index.Although this six can be measured
Index, but there is the disadvantages of detection speed is slow, and testing cost is high.And six indexs are detected respectively there is inconvenient,
The realistic problems such as error probability is big.And the HPLC (liquid chromatogram) most authoritative as traces component measure is although method result accurately may be used
It leans on, but cost is high, equipment is expensive, can not use on a large scale.In pork whether containing antibiotic, can safe edible be public
Eager inquisitive problem, in order to accelerate the speed of detection, many enterprises, testing agency, law enforcement agency, government have to spend
More human and material resources, time are respectively used to detection CLEN, RAC, SAL, CAP, CPZ, AMC and cause biggish waste.
Summary of the invention
In view of the above-mentioned problems existing in the prior art, the applicant provides a kind of eggs for detection of veterinary drugs in food
White chip and preparation method thereof.Protein chip of the present invention is only using our company for detecting six residue of veterinary drug indexs in pork
The protein chip technology of wound realizes that joint-detection can effectively improve detection efficiency, reduces testing cost；The present inventor's exploitation
Simultaneously include the protein chip diagnostic kit of CLEN, RAC, SAL, CAP, CPZ, AMC six indexs, has quickly, it is high
Effect, it is inexpensive the advantages that.
Technical scheme is as follows:
A kind of protein chip for detection of veterinary drugs in food, the preparation method of the protein chip include the following steps:
(1) black slide pretreatment；
(2) antigenic solution point sample；
(3) protein chip is made in closing process.
The black pretreated method of slide described in step (1) are as follows:
1. by black slide be placed in the slide pretreatment fluid containing NaOH impregnate 16~for 24 hours, later using purified water cleaning 2
~8 times；
2. black slide is placed in the solution of silane that mass concentration is 0.05~1% and impregnates 20~60min；
3. 0.2 is toasted under the conditions of 100~180 DEG C by being put into baking oven after soaked black slide nitrogen purging~
0.6h。
Antibody-solutions described in step (2) include CLEN antigenic solution, RAC antigenic solution, SAL antigenic solution, CAP antigen
Solution, CPZ antigenic solution, AMC antigenic solution.
The method of point sample described in step (2) is Machine automated point sample.
Closed process described in step (3) are as follows: submerge point sample good black slide 1 in confining liquid~for 24 hours, it takes out later black
Slide, and it is centrifuged the remaining confining liquid of removal, the protein chip is made.
The confining liquid is the buffer solution containing closed protein；The closed protein is bovine serum albumin(BSA) or egg white egg
It is white；The buffer is one of PBS buffer solution, Tris buffer, HEPS buffer, MOPS buffer or a variety of.
A kind of application of the protein chip, is made kit for the protein chip.
The kit further includes being marked with HRP enzyme or the corresponding antibody of secondary antibody solution, veterinary drug of alkali phosphorus enzyme is molten
Liquid, the chemiluminescent substrate sensitive to marker.
The present invention is beneficial to be had the technical effect that
Protein chip of the present invention realizes joint-detection for detecting six important veterinary drug indexs, using protein chip technology
Detection efficiency can be effectively improved, testing cost is reduced；It simultaneously include CLEN, RAC, SAL, CAP, CPZ, AMC six fingers
Target protein chip diagnostic kit, have quickly, efficiently, it is inexpensive the advantages that.Cooperate the automation protein chip of our company
Automatic detection may be implemented in reading apparatus.Since effectively six index integrations being detected in a chip, it is only necessary to
A pork sample quickly detects while can realizing six indexs.This product and technology are as a kind of novel detection side
Method, there are no similar products to appear on the market in the world at present.
The present invention is using classical immunology indirect competitive.Fixed trapped is anti-in the chip matrix that glass is carrier
Original, the specific antibody solution that these envelope antigens can be added with antigenic competition in sample, captured antigen-antibody are compound
Object is combined with the secondary antibody for being marked with HRP enzyme, forms sandwich combination product.It is added to marker (HRP enzyme or alkali phosphorus
Enzyme) sensitive chemiluminescent substrate carries out chemiluminescence, and optical signal passes through the power of optical signal by CCD camera acquisition
It may determine that the concentration of specific antigen in tested sample.
This kit has used residue of veterinary drug chip technology platform, and what the individual event detection method of other domestic producers used
It is ELISA detection method, an index can only be detected every time.In contrast, the advantage of this product is: 1. due to antigen or anti-
Body and this product antibody or the specificity of antigen binding is high, affinity is strong, and influenced by other impurities it is lower, it is therefore, right
The requirement of biological sample is very low, can simplify the pretreatment process of sample；And classical HPLC (liquid chromatogram) method to sample at
Reason requires very high.2. being capable of fast high-flux, a large amount of protein example of parallelization quantitative analysis；3. it is easy to operate, as a result just
True rate is high；4. required reagent and sample are few, cheap.
Detailed description of the invention
Fig. 1 is antigen point sample schematic diagram of the present invention；
In figure, column 1: positive quality control column；Column 2: blank；Arrange 3:CLEN measurement column；Arrange 4:RAC measurement column；Arrange 5:SAL measurement
Column；Column 6: blank；Arrange 7:CAP measurement column；Arrange 8:CPZ measurement column；Arrange 9:AMC measurement column；Column 10: blank；Column 11: negative control
Column.
Specific embodiment
With reference to the accompanying drawings and examples, the present invention is specifically described.
Embodiment 1
A kind of protein chip for detection of veterinary drugs in food, the preparation method of the protein chip include the following steps:
(1) black slide pretreatment；
1. black slide is placed in the slide pretreatment fluid containing 2%NaOH and impregnates 16h, later using purified water cleaning 2
~8 times；
2. black slide is placed in the solution of silane (medium is 25% ethyl alcohol) that mass concentration is 0.05% and impregnates 60min；
3. toasting 0.2h under the conditions of 180 DEG C for being put into baking oven after soaked black slide nitrogen purging.
(2) antigenic solution point sample；
It is anti-using Machine automated point sample CLEN antigenic solution, RAC antigenic solution, SAL antigenic solution, CAP with reference to Fig. 1
Original solution, CPZ antigenic solution, AMC antigenic solution, antigen solution concentration 0.05mg/mL, every point sample 20nL, each antibody point
Sample distribution is as shown in Figure 1.
(3) closing process；
The good black slide of point sample is submerged into 10h in confining liquid (PBS buffer solution for including 1% bovine serum albumin(BSA)), later
Black slide is taken out, and is centrifuged the remaining confining liquid of removal, the protein chip is made.
(4) sample process
A. the tissue samples after weighing 2.0 ± 0.05g homogeneous are into 50ml polystyrene centrifuge tube；
B. 1ml 1M HCL oscillation is first added to mix, it is mixed to add 6ml ethyl acetate-second eyeball solution oscillator oscillation
It is even, after mixing, (20-25 DEG C) centrifugation 10min of 4000g room temperature；
C. take 3ml supernatant in another clean 10ml centrifuge tube, nitrogen is blown in 60 DEG C of water-baths, until drying is
Only；
D. 1ml n-hexane is added into 10ml centrifuge tube, oscillation mixes；It adds 1ml and redissolves liquid, oscillation mixes, and mixes
Afterwards, (20-25 DEG C) centrifugation 10min of 4000g room temperature；
E. the upper liquid after removing centrifugation takes 100ul subnatant for detecting.
(5) kit
By the protein chip, residual animal medicine corresponding antibodies solution be marked with the secondary antibody solution (concentration of HRP enzyme
For 1ug/ml, wherein medium is the ELIAS secondary antibody dilution of the Sai Mofei company of outsourcing, pH=6.0), Sample dilution (0.01M
PBS PH-7.4), detection liquid A (containing 1% luminol and 2%Tris) and detection liquid B (containing 1% hydrogen peroxide) be packaged into examination jointly
Agent box.
Embodiment 2
A kind of protein chip for detection of veterinary drugs in food, the preparation method of the protein chip include the following steps:
(1) black slide pretreatment；
It is impregnated for 24 hours 1. being placed in black slide in the slide pretreatment fluid containing 2%NaOH, later using purified water cleaning 2
~8 times；
2. black slide is placed in the solution of silane (medium is 25% ethyl alcohol) that mass concentration is 0.5% and impregnates 30min；
3. toasting 0.5h under the conditions of 140 DEG C for being put into baking oven after soaked black slide nitrogen purging.
(2) antigenic solution point sample；
It is anti-using Machine automated point sample CLEN antigenic solution, RAC antigenic solution, SAL antigenic solution, CAP with reference to Fig. 1
Original solution, CPZ antigenic solution, AMC antigenic solution, antigen solution concentration 0.05mg/mL, every point sample 20nL, each antibody point
Sample distribution is as shown in Figure 1.
(3) closing process；
The good black slide of point sample is submerged in confining liquid (PBS buffer solution for including 2% ovalbumin) for 24 hours, is taken out later
Black slide, and it is centrifuged the remaining confining liquid of removal, the protein chip is made.
(4) sample process
A. the tissue samples after weighing 2.0 ± 0.05g homogeneous are into 50ml polystyrene centrifuge tube；
B. 1ml 1M HCL oscillation is first added to mix, it is mixed to add 6ml ethyl acetate-second eyeball solution oscillator oscillation
It is even, after mixing, (20-25 DEG C) centrifugation 10min of 4000g room temperature；
C. take 3ml supernatant in another clean 10ml centrifuge tube, nitrogen is blown in 60 DEG C of water-baths, until drying is
Only；
D. 1ml n-hexane is added into 10ml centrifuge tube, oscillation mixes；It adds 1ml and redissolves liquid, oscillation mixes, and mixes
Afterwards, (20-25 DEG C) centrifugation 10min of 4000g room temperature；
E. the upper liquid after removing centrifugation takes 100ul subnatant for detecting.
(5) kit
By the protein chip, residual animal medicine corresponding antibodies solution be marked with the secondary antibody solution (concentration of HRP enzyme
For 1ug/ml, wherein medium is the ELIAS secondary antibody dilution of the Sai Mofei company of outsourcing, pH=6.0), Sample dilution (0.01M
PBS PH-7.4), detection liquid A (containing 1% luminol and 2%Tris) and detection liquid B (containing 1% hydrogen peroxide) be packaged into examination jointly
Agent box.
Embodiment 3
A kind of protein chip for detection of veterinary drugs in food, the preparation method of the protein chip include the following steps:
(1) black slide pretreatment；
1. black slide is placed in the slide pretreatment fluid containing 2%NaOH and impregnates 20h, later using purified water cleaning 2
~8 times；
2. black slide is placed in the solution of silane (medium is 25% ethyl alcohol) that mass concentration is 1% and impregnates 20min；
3. toasting 0.6h under the conditions of 100 DEG C for being put into baking oven after soaked black slide nitrogen purging.
(2) antigenic solution point sample；
It is anti-using Machine automated point sample CLEN antigenic solution, RAC antigenic solution, SAL antigenic solution, CAP with reference to Fig. 1
Original solution, CPZ antigenic solution, AMC antigenic solution, antigen solution concentration 0.05mg/mL, every point sample 20nL, each antibody point
Sample distribution is as shown in Figure 1.
(3) closing process；
The good black slide of point sample is submerged into 8h in confining liquid (the Tris buffer for including 3% bovine serum albumin(BSA)), later
Black slide is taken out, and is centrifuged the remaining confining liquid of removal, the protein chip is made.
(4) kit assembles
By the protein chip, residual animal medicine corresponding antibodies solution be marked with the secondary antibody solution (concentration of HRP enzyme
For 1ug/ml, wherein medium is the ELIAS secondary antibody dilution of the Sai Mofei company of outsourcing, pH=6.0), Sample dilution (0.01M
PBS PH-7.4), detection liquid A (containing 1% luminol and 2%Tris) and detection liquid B (containing 1% hydrogen peroxide) be packaged into examination jointly
Agent box.
Test case:
Pork sample process
A. the tissue samples after weighing 2.0 ± 0.05g homogeneous are into 50ml polystyrene centrifuge tube；
B. 1ml 1M HCL oscillation is first added to mix, it is mixed to add 6ml ethyl acetate-second eyeball solution oscillator oscillation
It is even, after mixing, (20-25 DEG C) centrifugation 10min of 4000g room temperature；
C. take 3ml supernatant in another clean 10ml centrifuge tube, nitrogen is blown in 60 DEG C of water-baths, until drying is
Only；
D. 1ml n-hexane is added into 10ml centrifuge tube, oscillation mixes；It adds 1ml and redissolves liquid, oscillation mixes, and mixes
Afterwards, (20-25 DEG C) centrifugation 10min of 4000g room temperature；
E. the upper liquid after removing centrifugation takes 100ul subnatant for detecting.
Above-mentioned residue of veterinary drug sample is detected using the SLXP-001 type biological chip reading apparatus that our company produces,
The course of work of SLXP-001 type biological chip reading apparatus is as follows:
For instrument automatic sucking 100ul sample to be tested into reaction cup, instrument automatic sucking takes 100ul antibody-solutions to reaction
In cup, mix, then protein chip made from the embodiment of the present invention is automatically put into sample to be tested reaction cup by instrument, and 30 DEG C
It is incubated for 40 minutes, subsequent instrument clamping jaw takes out chip, and the secondary antibody for being marked with HRP enzyme is put into after instrument auto-flushing
Solution (200ul, instrument are inhaled in advance automatically), is incubated for after forty minutes again, and instrument clamping jaw takes out chip again, certainly through instrument
It puts into luminous substrate solution and (is mixed by the detection liquid B of detection the liquid A and 100ul of 100ul, by instrument after dynamic flushing
Automatic sucking and mixing), imaging of taking pictures finally is carried out to protein chip, software automatically analyzes picture, provides analysis result.Detection
As a result as shown in table 1, table 2, reference value is the single index measurement result of HPLC.
Table 1
Table 2
As seen from the above table, kit provided by the present invention, while detecting CLEN, RAC, SAL, CAP, CPZ, AMC six
Index, can obtain with similar in HPLC measurement result as a result, oneself sentencing since enterprise, testing agency, law enforcement agency, government have
The standard of disconnected yin and yang attribute, judgement of the present invention without yin and yang attribute.High efficiency may be implemented using this kit, letter operation is low
Cost, multiple advantages such as the used time is short are remarkably contributing in time, accurately by whether which kind of containing in testing result discovery pork
Antibiotic timely performs corresponding processing.
Claims (8)
1. a kind of protein chip for detection of veterinary drugs in food, it is characterised in that the preparation method of the protein chip includes as follows
Step:
(1) black slide pretreatment；
(2) antigenic solution point sample；
(3) protein chip is made in closing process.
2. protein chip according to claim 1, it is characterised in that the black pretreated method of slide described in step (1)
Are as follows:
1. by black slide be placed in the slide pretreatment fluid containing NaOH impregnate 16 ~ for 24 hours, later using purified water clean 2 ~ 8 times；
2. black slide is placed in the solution of silane that mass concentration is 0.05 ~ 1% and impregnates 20 ~ 60min；
3. toasting 0.2 ~ 0.6h under the conditions of 100 ~ 180 DEG C for being put into baking oven after soaked black slide nitrogen purging.
3. protein chip according to claim 1, it is characterised in that antigenic solution described in step (2) includes CLEN antigen
Solution, RAC antigenic solution, SAL antigenic solution, CAP antigenic solution, CPZ antigenic solution and AMC antigenic solution.
4. protein chip according to claim 1, it is characterised in that the method for point sample described in step (2) is that machine is automatic
Change point sample.
5. protein chip according to claim 1, it is characterised in that closed process described in step (3) are as follows: point sample is good
Black slide submerge 1 in confining liquid ~ for 24 hours, take out black slide later, and be centrifuged the remaining confining liquid of removal, the albumen core be made
Piece.
6. protein chip according to claim 5, it is characterised in that the confining liquid is that the buffering containing closed protein is molten
Liquid；The closed protein is bovine serum albumin(BSA) or ovalbumin；The buffer is PBS buffer solution, Tris buffer, HEPS
One of buffer, MOPS buffer are a variety of.
7. a kind of application of any one of claim 1 ~ 6 protein chip, it is characterised in that examination is made in the protein chip
Agent box.
8. application according to claim 7, it is characterised in that the kit further includes being marked with HRP enzyme or alkali phosphorus enzyme
The corresponding antibody-solutions of two antibody-solutions, veterinary drug, the chemiluminescent substrate sensitive to marker.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101231293A (en) * | 2008-02-14 | 2008-07-30 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Method for detecting vestigial clenbuterol |
| CN101782580A (en) * | 2009-01-16 | 2010-07-21 | 上海裕隆生物科技有限公司 | Protein chip for detecting multiple veterinary drug residues and kit thereof |
| CN102012424A (en) * | 2009-09-08 | 2011-04-13 | 宁波博奥生物工程有限公司 | Chip kit for quantitative detection of veterinary drug residue |
| CN103257233A (en) * | 2013-05-31 | 2013-08-21 | 南京祥中生物科技有限公司 | Biochip and method capable of simultaneously visually detecting multiple antibiotics, illegal addition agents and biotoxins |
| CN204347031U (en) * | 2015-01-19 | 2015-05-20 | 郑州大学 | The detection dish of 9 kinds of objectionable impuritiess in feed is detected fast the while of a kind of |
| CN106645060A (en) * | 2016-11-24 | 2017-05-10 | 福建蓝昊生物科技有限公司 | High-throughput veterinary drug residue detection protein antibody chip and application thereof |
| CN108519480A (en) * | 2018-03-13 | 2018-09-11 | 华南农业大学 | A kind of veterinary drug based on smart mobile phone remains immune detection system and detection method more |
| CN108828234A (en) * | 2018-08-21 | 2018-11-16 | 江苏三联生物工程有限公司 | A kind of protein chip and preparation method thereof for autoimmunity disease marker detection |
-
2019
- 2019-01-10 CN CN201910022566.7A patent/CN109613272A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101231293A (en) * | 2008-02-14 | 2008-07-30 | 中国人民解放军军事医学科学院卫生学环境医学研究所 | Method for detecting vestigial clenbuterol |
| CN101782580A (en) * | 2009-01-16 | 2010-07-21 | 上海裕隆生物科技有限公司 | Protein chip for detecting multiple veterinary drug residues and kit thereof |
| CN102012424A (en) * | 2009-09-08 | 2011-04-13 | 宁波博奥生物工程有限公司 | Chip kit for quantitative detection of veterinary drug residue |
| CN103257233A (en) * | 2013-05-31 | 2013-08-21 | 南京祥中生物科技有限公司 | Biochip and method capable of simultaneously visually detecting multiple antibiotics, illegal addition agents and biotoxins |
| CN204347031U (en) * | 2015-01-19 | 2015-05-20 | 郑州大学 | The detection dish of 9 kinds of objectionable impuritiess in feed is detected fast the while of a kind of |
| CN106645060A (en) * | 2016-11-24 | 2017-05-10 | 福建蓝昊生物科技有限公司 | High-throughput veterinary drug residue detection protein antibody chip and application thereof |
| CN108519480A (en) * | 2018-03-13 | 2018-09-11 | 华南农业大学 | A kind of veterinary drug based on smart mobile phone remains immune detection system and detection method more |
| CN108828234A (en) * | 2018-08-21 | 2018-11-16 | 江苏三联生物工程有限公司 | A kind of protein chip and preparation method thereof for autoimmunity disease marker detection |
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