CN105137068B - Florfenicol Site Detection reagent paper and its preparation, using method - Google Patents
Florfenicol Site Detection reagent paper and its preparation, using method Download PDFInfo
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- CN105137068B CN105137068B CN201510444444.9A CN201510444444A CN105137068B CN 105137068 B CN105137068 B CN 105137068B CN 201510444444 A CN201510444444 A CN 201510444444A CN 105137068 B CN105137068 B CN 105137068B
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- G—PHYSICS
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
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- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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Abstract
Present invention relates particularly to Site Detection reagent paper and its preparation, using method, belong to field of immunology.Including：Preserving number is the anti-florfenicol amine secreted by CCTCC C201575 hybridoma FFA C, monoclonal antibody C of colloid gold label, carrier board with adhesive sticker, the two ends of carrier board are provided with sample-adding end water accepting layer and handheld terminal water accepting layer, it is provided with detection layers in the middle part of this carrier board, it is provided with the glass layer block being loaded with gold mark monoclonal antibody C in this detection layers with sample-adding end water accepting layer intersection, one end of glass layer block is arranged under the water accepting layer of sample-adding end, the other end is arranged on detection layers, the detection layers continuing with glass layer block are provided with detection line and nature controlling line, it is coated with anti-florfenicol amine polyclonal antibody and goat anti-mouse igg respectively at detection line and nature controlling line.The present invention can scene be quick, easy, accurately detect florfenicol and its metabolite florfenicol amine, can be used for the scalping detection to florfenicol drug residue for the scene.
Description
Technical field
The present invention relates to the improvement of antibiotic detection technique is and in particular to Site Detection test strips and its preparation, user
Method, belongs to field of immunology.
Background technology
Florfenicol (Florfenico, FF) Chinese:Fluprofen, Florfenicol, chemical name:D (+)-Soviet Union -1-
P-methylphenyl -2- dichloro acetamino -3- fluorine propanol, is the later stage 1980s by U.S. Schering-
A kind of broad spectrum antibiotic of the special chloromycetin of beasts that Plough succeeds in developing, is third generation chloromycetin series antibioticses, this medicine
Nineteen ninety, as the succedaneum of new chloromycetin, the combination of main suppression 50s subunit was thus suppress bacterial peptide first in Japan's listing
The activity of acyl group transaminase, is the wide spectrum antibiosis of new resisting gram-positive bacteria, gram negative bacteria and thiamphenicol fastbacteria
Element, can be used for treating pig, the bacterial disease of cattle, fowl and aquatic animal.
Chloromycetin (Chloramphenicol, CAP) is as first generation chloromycetin series antibioticses ,-NO2 base in its structure
Group is relevant with suppression hemopoietic function, and therefore CAP has spinal cord hemopoietic function suppression toxicity, and reversible hemocyte can be caused to subtract
Few, the probability causing irreversible aplastic anemia is 1:30000.FF-CH3SO2 substituent group-NO2 group, no latent
Dangerous in aplastic anemia.The Resistant strain to resistance to chloromycetin and thiamphenicol (Thiamphenicol, TAP) for the FF simultaneously
Still there is antibacterial action.Florfenicol to play biological effect as targeted drug, the little instant effect of dosage, Increased Plasma Half-life,
Blood drug level is high, can maintain blood medicine for a long time, be widely used in prevention and the control of Animal diseases.China ratifies fluorobenzene Buddhist nun within 2000
Examine as national two class novel chiral synthon, aquaculture can be used for treat anguilla japonica Ai Dehuashi disease and red fin fish disease.
FF metabolism is very fast in animal body, and main metabolites are florfenicol oxamidic acid., florfenicol amine
(Florenicol amine, FFA), florfenicol alcohol etc., but FFA is topmost metabolism in most of edible animal tissue
The mark residue that product, therefore FFA were calculated as the off-drug period.Although Guo Guifang etc. thinks that FF injection is no significantly acute
Toxicity and subchronic toxicity, but there are some researches show that the FF of high dose can be with the weight reduction of mouse thymus, spleen, and external grinds
Study carefully humoral immunization and the cellular immunization also indicating that FF can suppress mice, the conclusion that this Guo Gui virtue draws mutually is confirmed, and shows FF
There is stronger immunotoxicity.FF can lead its residual in animal food in the extensive application in aquaculture, in order to ensure
Public health, China the maximum residue limit of florfenicol in animal tissue has been done regulation it is desirable to muscle residue limits be 200
μ g/Kg, liver residue limits are 3000 μ g/Kg, kidney residue limits are 300 μ g/Kg.
Content of the invention
The present invention is directed to the present situation of fluoride protector, and its harm to human body, and design invention offer one kind can
Scene is quick, easy, accurately detect florfenicol and the Test paper of its metabolite florfenicol amine and its preparation, user
Method.
Technical scheme is realized as follows
Florfenicol Site Detection reagent paper, its be characterized in that including：Preserving number is CCTCC C201575 hybridoma
Anti- florfenicol amine secreted by cell FFA-C, colloid gold label monoclonal antibody C (referred to as：Gold mark monoclonal antibody C), anti-fluorobenzene
Buddhist nun examines amine polyclonal antibody (referred to as：Multi-resistance) and goat anti-mouse igg is (referred to as：Sheep anti-mouse igg), the carrier board 3 with adhesive sticker,
At the both ends of carrier board 3, it is respectively equipped with sample-adding end water accepting layer 1 and handheld terminal water accepting layer 7, is provided with this carrier board 3 middle part
The detection layers 6 being made up of nitrocellulose membrane, are provided with sample-adding end water accepting layer 1 intersection in this detection layers 6 and are loaded with gold mark monoclonal antibody C's
Glass layer block 2, one end of glass layer block 2 is arranged under sample-adding end water accepting layer 1, and the other end is arranged on detection layers 6
On, detection line 4 and nature controlling line 5 are provided with the detection layers 6 continuing with glass layer block 2, at detection line 4 and nature controlling line 5
It is coated with multi-resistance and sheep anti-mouse igg respectively；
Described gold mark monoclonal antibody C is to prepare florfenicol amine (FFA) hapten and bovine serum albumin (BSA) by alkali hydrolysis method
It is coupled florfenicol amine-bovine serum albumin (FFA-BSA) complete antigen of synthesis by glutaraldehyde method, as immunogen
And the anti-florfenicol amine monoclonal antibody prepared, cause after colloid gold label；
Described anti-florfenicol amine polyclonal antibody：It is that florfenicol amine (FFA) hapten and ovum are prepared by alkali hydrolysis method
Pure albumen (OVA) is coupled complete antigen synthesis (FFA-OVA) by glutaraldehyde method and prepares as immunogen immune rabbit
Rabbit anteserum.
The preparation process of monoclonal antibody C of described colloid gold label is as follows：
1st, colloidal gold solution is prepared with the preparation technology of known gold colloidal, the granular size of prepared gold colloidal is 10-
20nm；
2nd, by monoclonal antibody C (0.5-1g/L antibody protein), it is added to above-mentioned 100ml gold colloidal by 150-500 μ l
In solution, mix even slowly；
3rd, add PEG20000 so as to final concentration of 1-5% in mixed solution, stand overnight；
4th, it is centrifuged 8000-10000 rev/min, 1-1.5 hour, abandon supernatant；
5th, take centrifugation, with containing 1-5%PEG20000,0.05-0.25% tween 20,0.02% sodium azide
0.01mol/L phosphate buffer hangs, and makes gold mark monoclonal antibody C；
The pH value of described phosphate buffer is 7.4, wherein contains KCI 0.2g, NaCI 8.0g, KH2PO40.2g、
Na2HPO412H2O 2.9g, distilled water 1000ml；
In described step 2, monoclonal antibody C and colloidal gold solution mix even 30-60min slowly.
The using method of florfenicol Site Detection reagent paper, it is characterized in that and comprises the following steps：
The sample-adding end water accepting layer 1 of this reagent paper is inserted or Deca is detected sample thereon, 5-10min is positioned at detection layers
The detection line 4 of 6 lower ends and show redness at the nature controlling line 5 positioned at detection layers 6 upper end, i.e. double red lines, representing residual in sample has
Florfenicol or florfenicol amine；No red line at detection line, represents that sample represents and can't detect florfenicol, florfenicol amine
Or both concentration are less than 10ug/ml, show redness at nature controlling line, represent that this reagent paper is effective.
It is an advantage of the current invention that：The present invention, with the actual features of prior art, have developed a kind of fluoride protector fast
Fast Site Detection reagent paper and its preparation, using method, add appropriate physiology salt water-powered roller using animal blood or internal organs such as liver
Broken be prepared into detection sample liquid, it is detected, principle is using sandwich immunoassay chromatographic theory, that is, by gold mark monoclonal antibody C with examining
Survey the antigen-reactive in sample liquid, formed by gold mark monoclonal antibody C- florfenicol or florfenicol amine compound, when this complex
When anti-florfenicol amine polyclonal antibody (FFA multi-resistance) place being coated in advance in detection line is chromatographed to nitrocellulose filter,
Antibody herein can identify antigen in this complex, and the result of reaction just defines gold mark monoclonal antibody C+ florfenicol or fluorobenzene Buddhist nun
Examine the sandwich structure of amine+FFA multi-resistance, be finally labelling on monoclonal antibody C colloid gold particle here fix and accumulate appearance naked eyes can
The red line seen, unreacted gold mark monoclonal antibody C then continues to chromatography and moves ahead, when the point of arrival is coated on nature controlling line sheep anti mouse in advance
When at IgG, Antibody types are that the golden standard gold mark monoclonal antibody C of IgG is combined with by it, thus also occur solid by colloid gold particle herein
Determine and accumulate and show macroscopic red line, result is：Detection line display redness indicates florfenicol or fluorobenzene
Buddhist nun examines amine residual, i.e. medicine setting sun；Detection line does not show redness, represents and can't detect florfenicol, florfenicol amine or two
Person's concentration is less than 10ug/ml, the i.e. residual feminine gender of medicine.Nature controlling line display is red, represents that reagent paper is effective；Redness is not shown at nature controlling line,
That is, or gold labeling antibody C or sheep anti-mouse igg inactivation illustrate that reagent paper lost efficacy, testing result is invalid.It is characteristic of the invention that：1、
The florfenicol Site Detection reagent paper of the present invention, from actual cultivation needs, the cultivation that the detection of drug residue is moved to
Scene, without specialized facilities and operating technology during use, is suitable for common raiser's cultivation site detection, application is simple.2nd, have
The features such as detection is quick, easy, accurate, sensitive, and there is higher stability, just can show detection knot within 5-10min
Really, once detection is exceeded can be allowed drug metabolism until residual is less than national standard foster a period of time more than cultivated animals, permissible
Safety lists, and is a kind of method of the residual problem of solution medicine of most convenient, decreases because drug residue is to the injury of human body and cultivation
Huge loss exceeded is caused because medicine is residual in family by the return of goods.Improve because of medicine residual censorship trouble, the cycle is long, and the reason such as charge height is made
Become the situation that more than 99% raiser is not censorship medicine residual before cultivated animals listing；3rd, anti-florfenicol amine prepared by this reagent paper
During multi-resistance, it is the multi-resistance of the complete antigen immunizing rabbit preparation being coupled with ovalbumin with florfenicol amine, this fundamentally keeps away
Exempt from the cross reaction with bovine serum albumin, the specificity further ensuring reagent paper is (because exempt from monoclonal antibody preparation
Epidemic focus is that have bovine serum albumin to be carrier protein)；4th, playing in whole colloid gold label and reagent paper preparation process prevents
Non-specific binding and the bovine serum albumin playing sealing process, are all replaced with the PEG20000 of debita spissitudo, are so entered one
Step has prevented the cross reaction that causes because of bovine serum albumin it is ensured that the specificity of test kit.
Brief description
Fig. 1：The structural representation of florfenicol Site Detection reagent paper of the present invention；
Fig. 2：The testing result schematic diagram of florfenicol Site Detection reagent paper of the present invention.
Hybridoma cell strain FFA-C, its deposit number is：CCTCC—C201575；Depositary institution's full name and abbreviation：In
State's Type Tissue Collection (CCTCC)；Depositary institution address：In the preservation of Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University
The heart；Preservation date：On May 21st, 2015.
Specific embodiment
Embodiment 1.
The synthesis of florfenicol amine：It is 1 according to mol ratio:1, weigh FF10.74, g sodium hydroxide 2.56g adds 15ml to steam
Distilled water dissolves, and to being completely dissolved, now solution is claret clarification to heated and stirred, adds 6.0g sodium chloride, stirs to complete
Dissolving, after cooling use thin layer chromatography plate layer chromatography mixed liquor, with iodine fumigate colour developing, record Rf (distance of initial point to spot centers and
Initial point is to the ratio of the distance of solvent front) value is different from FFC, proves that FFA generates.By solution left at room temperature over night, with filter
Paper filters, distilled water wash to trickle for colourless, be dried after FFA hapten, synthetic route is as follows：
Embodiment 2.
The preparation of complete antigen：Weigh 25mg florfenicol amine to be dissolved in 2mLDMF；Weigh 66mg BSA to be dissolved in
In the PBS of 10mlPH7.4；Florfenicol amine is added dropwise in above-mentioned BSA solution, stirs under the conditions of 4 DEG C, then
It is added dropwise over 100 μ L glutaraldehydes in reactant liquor, be slowly stirred reaction 24h.By the product PBS 72h of synthesis, every 8h changes
Liquid once, obtains florfenicol amine-bovine serum albumin (FFA-BSA) immunogen, prepares florfenicol amine-oralbumin with method
(FFA-OVA) coating antigen, synthetic route is as follows：
Embodiment 3.
The preparation of the monoclonal antibody of anti-florfenicol amine：The monoclonal antibody of the anti-florfenicol amine of secretion of the present invention
Hybridoma, its preparation is as follows with the method selecting：
(1) alkali hydrolysis method prepares florfenicol amine (FFA) hapten
(2) synthesize florfenicol amine complete antigen with glutaraldehyde method, including immunogen FFA-BSA, coating antigen FFA-OVA；
(3) use immunogen FFA-BSA immunity Balb/c white mice；
(4) by the fusion of the splenocyte of immune mouse and myeloma cell, 120 strain of hybridoma are cultivated to obtain；
(5) coating antigen FFA-OVA wrapper sheet indirect elisa method is used to screen positive hybridoma cell strain；
(6) using limiting dilution assay, wherein 4 plants of positive hybridoma cells are cloned；
(7) monoclonal antibody 2D6-1C4 bis- time cloning of the anti-florfenicol amine of potency highest is filtered out by indirect elisa method
The antibody of cell strain preparation, preparation gold mark monoclonal antibody.It is shown in Table 1
Table 1：Monoclonal antibody the selection result
Embodiment 4.
The specificity identification of the florfenicol amine monoclonal antibody C of the present invention：Different from 4 kinds with anti-florfenicol amine monoclonal antibody
Antibiotic be at war with ELLIA detection, use FFA-OVA wrapper sheet.Negative control PBS wrapper sheet；Sun contrasts, and is replaced with PBS using
Antibiotic in detection cross reaction.Testing result, florfenicol amine monoclonal antibody can be reacted with florfenicol amine and also can
React with florfenicol, both zero differences.And florfenicol amine monoclonal antibody and similar antibiotic thiamphenicol and other
The oxytetracycline of class such as Macrolide, the norfloxacin hydrochloride of quinolones and the equal no cross reaction of the sulfamethoxazole of sulfonamides.Say
This antibody specificity bright is strong, and can not only detect that florfenicol active compound also can detect its internal metabolite florfenicol
Amine.It is shown in Table 2
Table 2：The experiment of anti-florfenicol amine monoclonal antibody specificity (N=6)
Note：*. compare with positive controls, OD value is substantially reduced, (P ＜ 0.01)
Embodiment 5
Florfenicol Rapid detection test strip, referring to accompanying drawing 1, it includes：Preserving number is CCTCC C201575 hybridoma
Anti- florfenicol amine secreted by cell FFA-C, colloid gold label monoclonal antibody C (referred to as：Gold mark monoclonal antibody C), anti-fluorobenzene
Buddhist nun examines amine polyclonal antibody (referred to as：Multi-resistance) and goat anti-mouse igg is (referred to as：Sheep anti-mouse igg), the carrier board 3 with adhesive sticker,
At the both ends of carrier board 3, it is respectively equipped with sample-adding end water accepting layer 1 and handheld terminal water accepting layer 7, is provided with this carrier board 3 middle part
The detection layers 6 being made up of nitrocellulose membrane, are provided with sample-adding end water accepting layer 1 intersection in this detection layers 6 and are loaded with gold mark monoclonal antibody C's
Glass layer block 2, one end of glass layer block 2 is arranged under sample-adding end water accepting layer 1, and the other end is arranged on detection layers 6
On, detection line 4 and nature controlling line 5 are provided with the detection layers 6 continuing with glass layer block 2, at detection line 4 and nature controlling line 5
It is coated with multi-resistance and sheep anti-mouse igg respectively.
Embodiment 6
The preparation process of monoclonal antibody C of colloid gold label is as follows：
(1) preparation of gold colloidal：10-20nm colloid gold particle prepare, by concentration be 0.01% chlorauric acid solution 250ml with
1% sodium citrate 6.65ml mixing, is heated to 100 degrees Celsius, makes colloidal gold solution, the pH value of this solution：Use 0.2M carbonic acid
Potassium is adjusted to 8.2, standby；
(2) preparation of gold mark monoclonal antibody C：
When anti-florfenicol amine monoclonal antibody concentration is 1g/L, the most suitable monoclonal antibody amount of 100ml colloid gold label is 200 μ l, by this
Anti- florfenicol amine monoclonal antibody amine is added in colloidal gold solution ratio, stirs 50min slowly, adds 5%PEG20000,4 DEG C overnight；
Then, by it at 4 DEG C, 8000r is centrifuged 60min；Take centrifugation, with containing containing 5%PEG20000,0.25% polysorbas20
0.01mol/L phosphate buffer (KCL 0.2g, NaCL8.0g, KH2PO40.2g, Na2HPO412H2O 2.9g, distilled water
1000ml, pH 7.4) hang, then plus sodium azide concentration is adjusted to 0.02%, that is, make gold mark monoclonal antibody C.
Embodiment 7.
The preparation of anti-florfenicol amine multi-resistance：The complete antigen (FFA-OVA) being coupled with ovalbumin with florfenicol amine
For immunogen immune rabbit, the complete antigen (FFA-BSA) being coupled with bovine serum albumin with florfenicol amine is coating antigen, enters
Row immune enzyme-linked sorbent assay (ELISA), records potency and reaches 1:When 100000, Culling heart blood separation serum, 4 DEG C save backup.
Embodiment 8.
The florfenicol Site Detection detection paper layer preparation method of the present invention be：With saturated ammonium sulfate method purify immunity
Rabbit anteserum, freeze-dried after, be prepared into 1mg/ml anti-florfenicol amine multi-resistance, be sprayed on nitrocellulose membrane with Membrane jetter
On, form detection line 4；Equally, by goat anti-mouse igg, it is prepared into 200 μ g/ml, is sprayed on nitrocellulose membrane with Membrane jetter
On, form nature controlling line 5.At a distance of 5mm, the nearly handheld terminal water accepting layer of nature controlling line 5, detection line 4 is closely loaded end water accepting layer to two lines.Under room temperature
Dry, then with pH 7.4, the 37 DEG C of closing 30min of 0.010.01mol/LPBS containing 5%PEG20000, PBS rinsing is dried.
Embodiment 9.
The minimum residual quantity of the florfenicol quick detection bar detection of the present invention is 10ug/ml.It is shown in Table 3
Table 3：Florfenicol Site Detection reagent paper sensitively measures
Note：++ represent that detection line color is aubergine, testing result is strong positive；+ represent that detection line color is redness, inspection
Surveying result is the positive；Represent that detection line is colourless, testing result is feminine gender.
Embodiment 10.
The using method of the florfenicol Rapid detection test strip of the present invention, detecting step is as follows：First, from the pond of cultivation
In take live fish one, take liver, by weight/volume (w/v) compares 1:2 phosphate-buffered adding normal saline or pH 6.0-8.0
Liquid, liver is pulverized, and is prepared into detection sample；Then, sample-adding end 1 water accepting layer of reagent paper is immersed in the detection sample of test tube
(liquid level must not exceed the 2/3 of absorbent paper 1), 5min reads result.
Positive findingses：Inspected layer assumes two red detection lines 4 and nature controlling line 5 about 6, representing that sample is residual in having has fluorine
Benzene Buddhist nun examines or florfenicol amine florfenicol；
Negative findings：Inspected layer 6 upper end assumes a red nature controlling line 5, represents that sample represents and can't detect fluorobenzene Buddhist nun
Examine, florfenicol amine or both concentration are less than 10ug/ml；
Testing result is invalid：In sample-adding 5 minutes, at inspected layer 6 upper end nature controlling line 5, no red line occurs, and represents that this reagent paper loses
Effect, testing result is invalid.(see accompanying drawing 2).
Claims (7)
1. florfenicol Site Detection reagent paper is it is characterised in that include：Preserving number is CCTCC-C201575 hybridoma FFA-
Anti- florfenicol amine secreted by C, monoclonal antibody C (referred to as gold mark monoclonal antibody C) of colloid gold label, many grams of anti-florfenicol amine
Grand antibody and goat anti-mouse igg, the carrier board (3) with adhesive sticker, at the both ends of carrier board (3), it is respectively equipped with sample-adding end
Water accepting layer (1) and handheld terminal water accepting layer (7), are provided with the detection layers (6) being made up of nitrocellulose membrane in the middle part of this carrier board (3),
It is provided with the glass layer block (2) being loaded with gold mark monoclonal antibody C, glass fibers in this detection layers (6) with sample-adding end water accepting layer (1) intersection
One end of dimension layer block (2) is arranged under sample-adding end water accepting layer (1), and the other end is arranged on detection layers (6), with glass fibers
The detection layers (6) that dimension layer block (2) continues are provided with detection line (4) and nature controlling line (5), and detection line (4) and nature controlling line (5) place are respectively
It is coated with anti-florfenicol amine polyclonal antibody and goat anti-mouse igg.
2. florfenicol Site Detection reagent paper according to claim 1 is it is characterised in that described gold mark monoclonal antibody C is by aqueous alkali
Solution is prepared florfenicol amine hapten and bovine serum albumin and is passed through glutaraldehyde method coupling complete antigen synthesis FFA-BSA and is used as
Immunogen and the anti-florfenicol amine monoclonal antibody prepared, prepare after colloid gold label.
3. florfenicol Site Detection reagent paper according to claim 1 is it is characterised in that many grams of described anti-florfenicol amine
Grand antibody is to prepare florfenicol amine hapten and oralbumin by alkali hydrolysis method to be coupled synthesis by glutaraldehyde method completely anti-
The rabbit anteserum that original work are prepared for immunogen immune rabbit.
4. florfenicol Site Detection reagent paper according to claim 1 is it is characterised in that described gold marks the preparation side of monoclonal antibody C
Method is as follows：
(1) colloidal gold solution is prepared with the preparation technology of known gold colloidal, the granular size of prepared gold colloidal is 10-
20nm；
(2) by monoclonal antibody C, it is added in above-mentioned 100ml colloidal gold solution by 150-500 μ l, mixes even slowly；
(3) add PEG20000 so as to final concentration of 1-5% in mixed solution, stand overnight；
(4) it is centrifuged 8000-10000 rev/min, 1-1.5 hour, abandon supernatant；
(5) take centrifugation, with containing 1-5%PEG20000,0.05-0.25% tween 20,0.02% Hydrazoic acid,sodium salt
0.01mol/L phosphate buffer hangs, and makes gold mark monoclonal antibody C.
5. according to the florfenicol Site Detection reagent paper described in claim 4 it is characterised in that gold colloidal described in step (1) is molten
The preparation method of liquid is to be mixed and heated gold chloride and trisodium citrate by certain proportioning to be prepared into colloidal gold solution.
6. according to the florfenicol Site Detection reagent paper described in claim 4 it is characterised in that monoclonal antibody C in step (2)
Slowly mix even 30-60min with colloidal gold solution.
7. the using method of florfenicol Site Detection reagent paper is it is characterised in that comprise the following steps：Sample-adding end by this reagent paper
Water accepting layer (1) insertion or thereon Deca be detected sample, 5-10min the detection line (4) positioned at detection layers (6) lower end and
Show red, i.e. double red lines positioned at nature controlling line (5) place of detection layers (6) upper end, representing residual in sample has florfenicol or fluorine
Benzene Buddhist nun examines amine；No red line at detection line, represents that sample represents that to can't detect florfenicol, florfenicol amine or both concentration low
In 10 μ g/ml, at nature controlling line, show redness, represent that this reagent paper is effective；Do not show redness at nature controlling line, represent that this reagent paper is invalid.
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| CN106771159B (en) * | 2016-12-01 | 2019-05-28 | 无锡艾科瑞思产品设计与研究有限公司 | A kind of Florfenicol Rapid detection test strip |
| CN106905428A (en) * | 2017-04-25 | 2017-06-30 | 江南大学 | A kind of synthetic method of florfenicol amine artificial antigen |
| CN109709258B (en) * | 2019-01-30 | 2021-07-20 | 武汉生物工程学院 | Method for detecting florfenicol total residues in edible tissues of pigs and application |
| CN109709257A (en) * | 2019-01-30 | 2019-05-03 | 武汉生物工程学院 | A kind of method of Florfenicol and metabolite florfenicol amine in detection pig urcine |
| CN110988350A (en) * | 2019-11-08 | 2020-04-10 | 南京市产品质量监督检验院 | Quantum dot test paper capable of quantitatively detecting florfenicol residual quantity in various matrixes |
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