CN108267603A - Diclofenac quantum dot immune chromatography detection card and detection method based on signal amplifying system - Google Patents
Diclofenac quantum dot immune chromatography detection card and detection method based on signal amplifying system Download PDFInfo
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- CN108267603A CN108267603A CN201810264154.XA CN201810264154A CN108267603A CN 108267603 A CN108267603 A CN 108267603A CN 201810264154 A CN201810264154 A CN 201810264154A CN 108267603 A CN108267603 A CN 108267603A
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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
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9486—Analgesics, e.g. opiates, aspirine
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2430/00—Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
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Abstract
The present invention relates to a kind of Diclofenac quantum dot immune chromatography detection cards and detection method based on signal amplifying system.The detection card includes liner plate, detection antigen release pad, quantum dot probe release pad, chromatographic film and water absorption pad；Antigen release pad is detected to include by oralbumin and the diclofenac coupled detection antigen formed；Quantum dot probe release pad includes the quantum dot probe for being marked with anti-oralbumin antibody；Chromatographic film is equipped with detection line and nature controlling line, and detection line fixes anti-Diclofenac antibody, and nature controlling line fixes oralbumin antigen.Detection antigen dissolves in detection architecture from release pad, forms ternary system immune complex with detection antibody and labelled antibody, generates fluorescent assay signal.Under the detection reference effect of nature controlling line, whether Diclofenac is contained in judgement sample according to the fluorescence signal in detection line, the signal strength and stability of detection is increased, improves detection sensitivity and quantitative precision.
Description
Technical field
The present invention relates to food and drug safety detection field more particularly to it is a kind of by immunochemistry detection technique be applied to protect
The detection of the illegal addition chemicals of strong product, Chinese patent drug is based especially on the Diclofenac quantum dot immune of signal amplifying system
Chromatography detection card and detection method.
Background technology
Entitled 2- [(2, the 6- bis- chloro- 3- aminomethyl phenyls) amino] benzoic acid of chemistry of Diclofenac, belongs to non-steroidal anti-inflammatory
Medicine, have it is anti-inflammatory, analgesia and refrigeration function, for rheumatic arthritis, ankylosing spondylitis, non-inflammatory arthralgia, arthritis,
Pain caused by nonarticular rheumatism, non-non-articular inflammatory, various neuralgias, cancer pain, post-traumatic pain and various inflammation
Generate heat caused by disease etc., but have certain side effect, clinically belong to prescription medicine, it is necessary to be taken under the guidance of doctor.
Rheumatic disease is using pain such as joint, muscle, soft tissue, nerves as cardinal symptom, and the course of disease is mostly in chronic and anti-
Recurrence is made.There is the tradition by " dietotherapy " or herbal treatment chronic disease in China, and in chemosynthesis medicine, there are many side effects
Background under, favor in recent years by market for health food (based on health liquor), the Chinese patent drug of rheumatic disease.
Diclofenac analgesic effect is rapid and cheap, then has the illegal producer illegal in health food, Chinese patent drug
Diclofenac is added, illegitimate benefits is tryed to gain to heighten the effect of a treatment.It is improper take Diclofenac there may be nausea, headache or allergy
The adverse reactions such as property fash, cause to damage to alimentary canal, liver and kidney, serious to even result in anaphylactic shock and acute kidney
Functional failure.Patient is ignorant and takes the health food containing Diclofenac for a long time, and harmfulness is very serious, it is necessary to be added
By force to the supervision and check of illegal addition Diclofenac product.At present, the confirmation method for detecting illegal addition Diclofenac is efficient
Liquid chromatogram, LC-MS detection.But these method equipment investments are big, operating cost is high, sample pretreatment is complicated, it is impossible to
Scene is detected, it is difficult on a large scale to illegal addition phenomenon expansion screening.
Existing Diclofenac rapid detection method is mainly chemical detection method and thin-layered chromatography detection method, detection
The needs that sensitivity and antijamming capability are improved.Immunological detection method is sensitive, special, quick and inexpensive, in environmental monitoring
It is had been widely used with field of food safety.The immunological method of detection micromolecular compound reported at present, is all based on detecting
Semiochemicals are marked on detection antibody, and in chromatographic film by " the binary system Immune competition method " of antigen and detection antibody
Fixed detection antigen forms immune complex, generates detection signal." binary system Immune competition method " has the following disadvantages：1、
It need to cause semiochemicals that can not unify to prepare each detection antibody difference marking signal substance；2nd, detection antibody is in liquid phase, surely
Qualitative deficiency；3rd, it detects signal strength and sensitivity is relatively low.
Invention content
Based on this, the object of the present invention is to provide a kind of Diclofenac quantum dot immunes based on signal amplifying system
Chromatography detection card and detection method have the advantages that increase detection signal strength and stability, improve detection sensitivity.
The purpose of the present invention is what is be achieved through the following technical solutions：
Diclofenac quantum dot immune chromatography detection card based on signal amplifying system, adheres to including liner plate and successively
In detection antigen release pad, quantum dot probe release pad, chromatographic film and the water absorption pad of overlapping portions on liner plate and between adjacent；Institute
Detection antigen release pad is stated to include by oralbumin and the diclofenac coupled detection antigen formed；The quantum dot probe is released
It puts pad and includes the quantum dot probe for being marked with anti-oralbumin antibody；The chromatographic film is equipped with detection line and nature controlling line, described
Detection line fixes anti-Diclofenac antibody, and the nature controlling line fixes oralbumin antigen.
Compared to traditional " binary system Immune competition method ", detection card of the invention is utilized that " ternary system is immunized competing
Strive method " principle, except " detection antigen " and " detection antibody " the two immune detection elements, increase and carried for detection antigen
" labelled antibody " of body protein.The present invention is covalently attached detection substance (Diclofenac) on carrier protein (oralbumin) and closes
Into detection antigen, labelled antibody (anti-oralbumin antibody) is marked on quantum dot, by detection antibody, (anti-Diclofenac resists
Body) it is fixed in chromatographic film, so as to detect the detection substance being coupled on antibody capture detection antigen, it is anti-that labelled antibody combines detection
The carrier protein being coupled in original forms double-antibody sandwich immune complex, generates fluorescent assay signal.When detection antibody is dissociated
Detection substance combine, fluorescence intensity will be suppressed, so as to generate inhibit detection signal.
Specifically, dripping to sample solution in detection antigen release pad during detection, sample solution will inspection during moving
Antigen and quantum dot probe dissolving are surveyed, and is carried to detection line and nature controlling line, the anti-egg white of quantum dot probe label being carried
Albumin (OVA) antibody, detection antigen and the anti-Diclofenac antibody in detection line form double-antibody sandwich immune conjugate, make
Detection line generates fluorescence signal；The anti ova antibody and fixed OVA antigens knot on nature controlling line for the quantum dot probe label being carried
It closes, is accumulated on nature controlling line and form fluorescence signal.Reach a certain concentration when there is free Diclofenac in sample, in detection line
Immune response will be blocked by competition, and fluorescence signal is suppressed；And the fluorescence signal on nature controlling line, not by Diclofenac concentration
It influences.
Relevant report currently with fluorescence quantum immune chromatography method detection Diclofenac is less, and the present invention is food
Drug safety detection work provides new tool.Quantum dot is the nanometer that three diameter of Spherical Volume are collected at by 200-10000 atoms
Semiconductor crystal, spherical nucleus diameter 2-8nm, in order to increase the water solubility of quantum dot and biocompatibility, spherical nucleus outer layer
It would generally be coated by organic molecule and introduce functional group, diameter can be increased to 15-30nm, have good colloidal property and power
Characteristic is learned, is suitable as the Nanoparticle labeling material of Biological Detection.Compared with the nanoparticle of traditional organic fluorescent dye filling,
Quantum dot has wide and is in continuously distributed excitation spectrum, narrow and symmetrical emission peak；Stability is strong, anti-light Bleachability strong；Luminous energy
Power transformation efficiency height, brightness are that the decades of times of organic dyestuff is even more.
Relative to the prior art, detection fixture of the invention has the advantage that：1st, it is uniformly marked with anti-carrier protein antibodies
Quantum dot generate detection signal, only need centralized system for a kind of as fluorescence probe, be conducive to detect scale and the mark of work
Standardization；2nd, detection antibody is fixed in chromatographic film, by the closing and drying to film, is more had to the active protection effect of antibody
Profit increases Antibody stability；3rd, steric hindrance smaller when combining is immunized, improves detection signal strength and sensitivity；4th, it is specific
By force, detection time is short (5-10 minutes), can execute-in-place, and can excite fluorescence signal by portable 360nm light sources, visually sentence
It reads as a result, testing cost is low, easy to operate, suitable base testing staff operates.
Further, the detection antigen release pad is absorbed using glass fibre element film containing detection antigen, surface-active
Agent, mannitol, sucrose PBS solution be made.Preferably, use thickness for the glass fibre element film of 0.85mm fully absorb containing
10 μ g/mL detect the PBS solution of antigen, 50 μ g/mL surfactants, 30mg/mL mannitol, 50mg/mL sucrose.Wherein, it is sweet
Dew alcohol is used to ensure that detecting antigen in detection process to dissolve rapidly as freeze-drying stent；Sucrose detects solution viscosity control for adjusting
Preparative layer analyses development rate；Surfactant is used to eliminate the non-specific adsorption of detection process, preferably polyethylene glycol octyl phenyl
Ether (Triton X-100)；Detection antigen is synthesized by OVA and Diclofenac covalent coupling, can detect antibody combination on envelope,
It can be combined again by the labelled antibody on quantum dot.
Further, the quantum dot probe release pad using artificial cellulose's film absorb containing quantum dot fluorescence probe,
Polyethylene glycol, mannitol, sucrose, glycine PBS solution be made.Preferably, artificial cellulose of the thickness for 0.34mm is used
Film is absorbed containing 20 μ g/mL quantum dot fluorescence probes, 60 μ g/mL polyethylene glycol (PEG-500), 10mg/mL mannitol, 60mg/
The PBS solution of mL sucrose, 10mg/mL glycine.Wherein, mannitol is used to ensure quantum dot in detection process as freeze-drying stent
Probe dissolves rapidly；Polyethylene glycol, sucrose chromatograph development rate for adjusting detection solution viscosity control；Glycine is used to eliminate
The non-specific adsorption of detection process；Quantum dot fluorescence probe is made of quantum dot surface label anti ova antibody, can combine quilt
The detection antigen in detection line and the OVA antigens being fixed on nature controlling line are captured, generates fluorescent assay signal respectively.
Further, the chromatographic film use nitrocellulose filter, the detection line by contain anti-Diclofenac antibody and
The PBS solution of sucrose is sprayed on nitrocellulose filter and is made.Preferably, will contain the anti-Diclofenac antibody of 0.3mg/mL and
The 0.05M PBS solutions (pH 7.4) of 10mg/mL sucrose, are sprayed on the amount of 1.0~4.0 μ g/cm on nitrocellulose filter.Inspection
The basis that survey line fluorescence signal is formed is the immune response of antibody and Diclofenac coupled by detection antigen, can be detected in product and dissociate
Diclofenac Competitive assays.
Further, the nature controlling line is sprayed on nitrocellulose filter by the PBS solution containing oralbumin and sucrose
It is upper to be made.Preferably, by the 0.05M PBS solutions (pH 7.4) containing 0.2mg/mL OVA and 10mg/mL sucrose, with 1.0~
The amount of 3.0 μ g/cm is sprayed on nitrocellulose filter.The basis that nature controlling line fluorescence signal is formed is the anti ova antibody of quantum dot
It is unrelated with Diclofenac inspection product with the immune response of Quality Control OVA antigens, the Diclofenac competition suppression to dissociate in product will not be detected
System.
Diclofenac quantum dot immune chromatography detection method based on signal amplifying system, includes the following steps：
S1：Prepare detection antigen, quantum dot probe, detection line and nature controlling line；The detection antigen by oralbumin and
Diclofenac coupled to form, the quantum dot probe is marked with anti-oralbumin antibody, and it is fragrant that the detection line fixes anti-double chlorine
Sour antibody, the nature controlling line fix oralbumin antigen；
S2：Sample solution, detection antigen and quantum dot probe are mixed, are delivered to detection line and nature controlling line together, according to
Fluorescence signal in detection line and nature controlling line comes in judgement sample whether contain Diclofenac.
Diclofenac is micromolecular compound, and the immune detection principle of conventional detection micromolecular compound is " detection antigen "
" the binary system Immune competition method " formed with " detection antibody ", for this method in the lateral immunochromatography of quantum dot not
Foot, the present invention add in " labelled antibody " formation " ternary system Immune competition on the basis of " detection antigen " with " detection antibody "
Method ", and combination of each immune response element in tomographic system is also adjusted, detection antibody is fixed on detection line,
Label anti ova antibody forms fluorescence probe over the qds, with detecting antigen (OVA- Diclofenacs) " bridging " formation " double antibody
It is sandwich " immune complex, fluorescence probe accumulates to form detection signal, increases the signal strength and stability of detection, improve
Detection sensitivity, when detection antibody is combined by free detection substance, fluorescence intensity will be suppressed, and inhibit detection letter so as to generate
Number.Be 2.0ng/mL to the minimum detectability degree of Diclofenac in sample solution using the detection method of the present invention, to health products,
The minimum detectability degree of illegal addition Diclofenac and the like is 2.0 μ g/kg, and can complete in 5 minutes in Chinese patent drug
Quick detection.
Further, in step S2, judged by the following method：Detection line and nature controlling line all show fluorescence signal,
Conclude that for feminine gender, Diclofenac is free of in sample for result；Detection line does not show fluorescence signal, and nature controlling line shows fluorescence signal, breaks
Result is determined for the positive, contains Diclofenac in sample.Wherein, nature controlling line be in order to the method for inspection effectively whether and set, Quality Control
Line shows that fluorescence signal shows that method is effective, and nature controlling line does not show that fluorescence signal shows that method is invalid in itself；And detection line is formed
Fluorescence signal being generated during double-antibody sandwich immune complex, when there is free Diclofenac in sample, being exempted from detection line
Epidemic disease reaction will be blocked by competition, so as to which fluorescence signal disappears.
Further, the quantum dot probe generates the transmitting light of 630nm under 365nm light source activations.
In order to better understand and implement, the invention will now be described in detail with reference to the accompanying drawings.
Description of the drawings
Fig. 1 is the structural representation of the chromatography detection card of the Diclofenac quantum dot immune based on signal amplifying system of embodiment
Figure.
Fig. 2 a are that immune response, the schematic diagram for generating fluorescence signal occur in detection line.
Fig. 2 b are the schematic diagram that immune response is blocked by competition, fluorescence signal disappears in detection line.
Fig. 3 is that immune response, the schematic diagram for generating fluorescence signal occur on nature controlling line.
Fig. 4 is the judgment principle of the principle that fluorescence immunoassay testing result generates and testing result.
Specific embodiment
Referring to Fig. 1, its chromatography detection of the Diclofenac quantum dot immune based on signal amplifying system for the present embodiment
Card is adhered to including PVC liner plates and successively on PVC liner plates and the detection antigen release pad of overlapping portions, quantum between adjacent
Point probe release pad, nitrocellulose filter (NC films) and water absorption pad；The detection antigen release pad is included by OVA and Diclofenac
It is coupled the detection antigen formed；The quantum dot probe release pad includes the quantum dot probe for being marked with anti ova antibody；The nitre
Acid cellulose film is equipped with detection line and nature controlling line, and the detection line fixes anti-Diclofenac antibody, and the nature controlling line is fixed OVA and resisted
It is former.
Specifically, the preparation method of detection antigen release pad is as follows：Thickness is used to be filled for the glass fibre element film of 0.85mm
Divide the PBS for absorbing and antigen, 50 μ g/mL Triton X-100,30mg/mL mannitol, 50mg/mL sucrose being detected containing 10 μ g/mL
Solution, freeze-dried back.
Specifically, the preparation method of quantum dot probe release pad is as follows：First, using active ester method to carboxylated quantum dot
It is marked, 1.0mL carboxylated quantum dots is taken to be diluted in 20mL ultra-pure waters, add in the EDC solution of 0.05mL Fresh
(10mg/mL), being stirred at room temperature 20 minutes makes activated carboxylic；0.45mg anti ova antibody is added in pre-activate microballoon, is stirred at room temperature
Connect 30 minutes the amino of activated carboxyl and antibody；Solution in 16000rpm low-temperature centrifugations 30 minutes, discard supernatant liquid with point
From unbonded antibody；Quantum dot probe is resuspended with the PBS solution of the X-100 containing 10%Triton, is protected from light 4 DEG C and is saved backup.So
Afterwards, thickness is used to be absorbed for 85 artificial cellulose's films of Whatman of 0.34mm containing 20 μ g/mL quantum dot fluorescence probes, 60 μ
G/mL PEG-500,10mg/mL mannitol, 60mg/mL sucrose, 10mg/mL glycine PBS solution, freeze-dried back.
Specifically, the preparation method of detection line is as follows：The anti-Diclofenac specific antibodies of 0.3mg/mL and 10mg/ will be contained
The 0.05M PBS solutions (pH 7.4) of mL sucrose, are sprayed on the amount of 1.00 μ g/cm on nitrocellulose filter, form detection line.
Specifically, the preparation method of nature controlling line is as follows：By the 0.05M containing 0.2mg/mL OVA and 10mg/mL sucrose
PBS solution (pH 7.4) is sprayed on the amount of 1.00 μ g/cm on nitrocellulose filter, forms nature controlling line.
Diclofenac and the like specific antibody induction production method, anti ova antibody induction production method and
Label anti ova antibody, the method for preparing quantum dot fluorescence probe use the prior art over the qds.
Diclofenac quantum dot immune based on the signal amplifying system chromatography detection method of the present embodiment is as follows：
0.2g (or 0.2mL) sample is taken, is dissolved in 2.0mL absolute ethyl alcohols and fully extracts target inspection product, extract is static
Impurity is made fully to precipitate within 10 minutes, 0.2mL supernatants is taken to be added to 20mL sample buffers, fully shaken up as sample solution；
3-4 drop sample solutions are added dropwise in detection antigen release pad with dropper, during sample solution is moved to nitrocellulose filter
Detection antigen and quantum dot probe are discharged, and successively crosses detection line and nature controlling line, according to glimmering in detection line and nature controlling line
Optical signal comes in judgement sample whether contain Diclofenac.
The anti ova antibody marked on anti-Diclofenac antibody, quantum dot probe in nitrocellulose filter detection line, point
Not with detecting antigen binding, double-antibody sandwich immune conjugate is formed, detection line is made to generate fluorescent assay signal, such as Fig. 2 a institutes
Show.Reach a certain concentration when there is free Diclofenac in sample, immune response will be by competition blocking, fluorescence in detection line
Signal can disappear, as shown in Figure 2 b.Quantum dot probe is by the anti ova antibody of label with being fixed on nitrocellulose filter nature controlling line
OVA antigen bindings, on nature controlling line accumulation form fluorescence signal, as shown in Figure 3.Nature controlling line be the method for inspection in itself effectively with
Not no and set, colour developing is effective, does not develop the color and shows that method is invalid in itself.
As shown in figure 4, quantum dot probe under 365nm light source activations, generates the transmitting light of 630nm, testing result is sentenced
Disconnected principle is：Detection line and nature controlling line all show fluorescence signal, and it is negative (A) to conclude result, and Diclofenac is free of in sample；Inspection
Survey line does not show fluorescence signal, and nature controlling line shows fluorescence signal, and it is positive (B and C) to conclude result, and it is fragrant that double chlorine are contained in sample
Acid, wherein, Diclofenac concentration is higher than 2.0ng/ in a concentration of 1.0ng/mL of Diclofenac in positive findings B, positive findings C
mL。
Relative to the prior art, " the ternary system Immune competition method " that the present invention uses has the following advantages：1st, it uniformly uses
The quantum dot of anti-carrier protein antibodies label generates detection signal, and centralized system is only needed, as fluorescence probe, to be conducive to examine for a kind of
Survey the scale and standardization of work；2nd, detection antibody is fixed in chromatographic film, by the closing and drying to film, to antibody
Active protection acts on advantageously, increases Antibody stability；3rd, steric hindrance smaller when combining is immunized, improves detection signal strength
And sensitivity.Using the detection method of the present invention to being 2.0ng/mL to the minimum detectability degree of Diclofenac in sample solution,
Minimum detectability degree to addition Diclofenac illegal in health products, Chinese patent drug and the like is 2.0 μ g/kg, and can be at 5 points
Quick detection is completed in clock.
Embodiment described above only expresses the several embodiments of the present invention, and description is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that those of ordinary skill in the art are come
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Range.
Claims (8)
1. the Diclofenac quantum dot immune chromatography detection card based on signal amplifying system, it is characterised in that：Including liner plate and
Be adhered to successively on liner plate and the detection antigen release pad of overlapping portions between adjacent, quantum dot probe release pad, chromatographic film and
Water absorption pad；The detection antigen release pad is included by oralbumin and the diclofenac coupled detection antigen formed；The amount
Sub- point probe release pad includes the quantum dot probe for being marked with anti-oralbumin antibody；The chromatographic film is equipped with detection line and matter
Line is controlled, the detection line fixes anti-Diclofenac antibody, and the nature controlling line fixes oralbumin antigen.
2. the Diclofenac quantum dot immune chromatography detection card according to claim 1 based on signal amplifying system, special
Sign is：The detection antigen release pad using glass fibre element film absorb containing detection antigen, surfactant, mannitol,
The PBS solution of sucrose is made.
3. the Diclofenac quantum dot immune chromatography detection card according to claim 1 based on signal amplifying system, special
Sign is：The quantum dot probe release pad is absorbed using artificial cellulose's film contains quantum dot fluorescence probe, polyethylene glycol, sweet
Dew alcohol, sucrose, glycine PBS solution be made.
4. the Diclofenac quantum dot immune chromatography detection card according to claim 1 based on signal amplifying system, special
Sign is：The chromatographic film uses nitrocellulose filter, and the detection line is molten by the PBS for containing anti-Diclofenac antibody and sucrose
Liquid is sprayed on nitrocellulose filter and is made.
5. the Diclofenac quantum dot immune chromatography detection card according to claim 4 based on signal amplifying system, special
Sign is：The nature controlling line is sprayed on nitrocellulose filter by the PBS solution containing oralbumin and sucrose and is made.
6. the Diclofenac quantum dot immune chromatography detection method based on signal amplifying system, it is characterised in that：Including following step
Suddenly：
S1：Prepare detection antigen, quantum dot probe, detection line and nature controlling line；The detection antigen is by oralbumin and double chlorine
Fragrant acid is coupled, and the quantum dot probe is marked with anti-oralbumin antibody, and the detection line is fixed anti-Diclofenac and resisted
Body, the nature controlling line fix oralbumin antigen；
S2：Sample solution, detection antigen and quantum dot probe are mixed, detection line and nature controlling line are delivered to together, according to detection
Fluorescence signal on line and nature controlling line comes in judgement sample whether contain Diclofenac.
7. the Diclofenac quantum dot immune chromatography detection method according to claim 6 based on signal amplifying system,
It is characterized in that：In step S2, judged by the following method：Detection line and nature controlling line all show fluorescence signal, conclude result
For feminine gender, Diclofenac is free of in sample；Detection line does not show fluorescence signal, and nature controlling line shows fluorescence signal, concludes that result is
The positive contains Diclofenac in sample.
8. the Diclofenac quantum dot immune chromatography detection method according to claim 6 based on signal amplifying system,
It is characterized in that：The quantum dot probe generates the transmitting light of 630nm under 365nm light source activations.
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| CN109884293A (en) * | 2019-03-29 | 2019-06-14 | 中国海洋大学 | A kind of food allergen rapid detection method based on quantum dot fluorescence |
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| WO1998023958A1 (en) * | 1996-11-27 | 1998-06-04 | Schleicher & Schuell, Inc. | Competitive lateral flow, specific binding chromatographic assay devices, kits, and methods |
| CN2515679Y (en) * | 2002-01-10 | 2002-10-09 | 河南省农业科学院生物技术研究所 | Clenbuterol hydrochloride fast detecting test paper strip (II) |
| CN2515678Y (en) * | 2002-01-10 | 2002-10-09 | 河南省农业科学院生物技术研究所 | Test paper strip for fast detecting drug residue of animal body and its product |
| CN1403815A (en) * | 2002-01-10 | 2003-03-19 | 河南省农业科学院生物技术研究所 | Fast detection test paper strip for medicine residue in animal body and product |
| CN101551398A (en) * | 2008-11-26 | 2009-10-07 | 中国计量学院 | Drug residue competition-type quantum dot-labeled immunochromatography assay test-strip and observation device thereof |
| CN102229672A (en) * | 2011-05-23 | 2011-11-02 | 重庆大学 | Single-chain antibody against fenitrothion and preparation method thereof |
| CN103267842A (en) * | 2013-05-15 | 2013-08-28 | 郭杰标 | Immune colloidal gold method for detecting diclofenac illegally added in Chinese patent medicament |
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| CN109884293A (en) * | 2019-03-29 | 2019-06-14 | 中国海洋大学 | A kind of food allergen rapid detection method based on quantum dot fluorescence |
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