CN108548925A - Ternary system Immune competition method detects the quantum dot immune chromatography detection card and detection method of silaenafil similar drug - Google Patents
Ternary system Immune competition method detects the quantum dot immune chromatography detection card and detection method of silaenafil similar drug Download PDFInfo
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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
- G01N33/9453—Cardioregulators, e.g. antihypotensives, antiarrhythmics
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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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to quantum dot immune chromatography detection cards and detection method that a kind of ternary system Immune competition method detects silaenafil similar drug.The detection card includes liner plate, detection antigen release pad, quantum dot probe release pad, chromatographic film and water absorption pad；Detection antigen release pad includes to detect antigen made of oralbumin and the coupling of silaenafil similar drug；Quantum dot probe release pad includes to be marked with the quantum dot probe of anti-oralbumin antibody；Chromatographic film is equipped with detection line and nature controlling line, and detection line fixes anti-silaenafil similar drug 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 silaenafil similar drug 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, especially ternary system Immune competition method detect silaenafil similar drug
Quantum dot immune chromatography detection card and detection method.
Background technology
In the phase at the beginning of the nineties in last century, a kind of PDE-5 inhibitor medicaments for treating coronary heart disease of Pfizer are facing
It finds effectively treat impotence in bed experiment, here it is sildenafil citrates, produce huge business profit on the market
Benefit.
It is driven by huge market interest, criminal's synthesis illegal addition silaenafil class medicine in natural establishing-Yang product
Object (including its various analogue) increases Yang-strengthening effect to promote to sell, seeks the behavior of unlawful interests.This behavior pair
The harm of public health has become troubling global problem.
The harmfulness of illegal addition silaenafil class drug is embodied in：First, PDE-5 inhibitor medicaments can cause a series of
Side effect, such as headache, flush, indigestion, eye-blurred and DOMS.The drug of U.S. FDA official Internet page publication
Adverse reaction presentation of information, taking silaenafil class drug may cause to blind, and also result in hearing and decline or even become deaf suddenly.
Therefore, illegal add is taken in countries in the world to silaenafil class drug all in strict accordance with prescription medicine management under no physician guidance
The similar drug added is dangerous.Secondly, the interaction of silaenafil class drug and nitrate drug can cause serious low
Blood pressure.Nitrate drug is widely used in diabetes, hypertension, the treatment of hyperlipidemia and coronary heart disease, with these diseases and
With the patient of impotence symptom, natural establishing-Yang drug is often sought help from.If these patients have taken nitrate drug,
Without knowing it, have and taken the Chinese patent drug (health products) containing illegal silaenafil class drug, it will it is fearful to cause
Consequence.
Criminal also carries out modification transformation to the structure of silaenafil, synthesized a series of sildenafil derivative into
The illegal addition of row.Cost is lower, effect becomes apparent from due to being added in health products, and silaenafil similar drug is in Chinese patent drug (health care
Product) in illegal addition the case where, have the tendency that more and more fiery.
Illegal additive capacity smaller, the concealment of some silaenafil analogs are stronger, lead to screening detection difficulty bigger,
The R＆D intensity of corresponding detection technique must be increased.And in recent years, the detection department of countries in the world and scientific research institution are that exploitation is disobeyed
Method PDE-5 drug screening methods have done a large amount of work, are mainly the following technology：1, thin-layer chromatography (TLC) technology.TLC
Technically simple easy, testing cost is cheap, is applicable in extensive Preliminary Identification.T.S.Reddy etc. is developed the color using bismuth potassium iodide, is established
High Performance Thin Layer Chromatography (HPTLC) technology, under the reference of standard items, the silaenafil class of illegal addition in screening natural medicine
Drug achieves good effect.2, high performance liquid chromatography (HPLC) and LC-MS (LC-MS) technology.High performance liquid chromatography
(HPLC) technology is widely used in the screening of silaenafil class drug, and maximum advantage can be achieved on quantitative detection, liquid
Matter combination (LC-MS) can carry out confirmation inspection in conjunction with the Information in Mass Spectra of inspection product to the silaenafil class drug of illegal addition.3、
HPLC and LC-MS can only detect known silaenafil, for molecular structure by the emerging silaenafil class of premeditated modification
Like object, since chromatographic behavior and mass spectrum behavior have differences with known drug, only liquid phase/tandem ion-trap mass spectrum (LC/ESI-
MS/MS) technology can detect.Inspection product isolated LC are converted to quasi-molecular ion by the technology with electron spray ionisation (ESI),
The characteristic ion fragment that dissociation generation is carried out through collision cell (CID) is detected in second order ms (MS2), and comprehensive analysis examines product liquid
The compound structure information that phase separation result and tandem mass spectrum obtain can detect known and unknown in same process
Silaenafil class drug.4, in addition to expensive LC/ESI-MS/MS, only pass through hydrolysising product analysis, thus it is speculated that new west ground occur
The molecular structure of that non-class drug.By carrying out acidolysis to inspection product, hydrolysate is analyzed with LC-MS, grasps the inspection product
Each hydrolysate column retention time and m/z values, detected with the LC-MS of the hydrolysate of known PDE-5 drug standards
As a result it is compared, can speculate the precision architecture of target inspection product.
Above instrument detection method is although sensitive, accurate, reliable, but equipment costly, service condition it is harsh, right
Personnel qualifications are high, cannot carry out Site Detection, and cracking down on counterfeit goods, there are limitations for detection working effect.Must have it is quick, sensitive,
Reliably, inexpensive in-situ check and test method is supplemented.
Existing silaenafil similar drug rapid detection method is mainly chemical detection method and thin-layered chromatography detection method,
The needs that the sensitivity of detection and anti-interference ability are improved.Immunological detection method is sensitive, special, quick and inexpensive, in ring
Border monitors and field of food safety has been widely used, and great expectations is also increasingly given in food and drug safety quickly detects.Mesh
Preceding report detection micromolecular compound immunological method, be all based on detection antigen and detect antibody " binary system is exempted from
Semiochemicals are marked on detection antibody, are formed with fixed detection antigen in chromatographic film immune compound by epidemic disease competition law "
Object generates detection signal." binary system Immune competition method " has the following disadvantages：1, letter need to be marked respectively to each detection antibody
Number substance causes semiochemicals that can not unify to prepare；2, for detection antibody in liquid phase, stability is insufficient；3, detect signal strength and
Sensitivity is relatively low.
In the previous work of the present invention, animal is immunized in the artificial antigen to protrude silaenafil similar drug common group, lures
Artificial delivery life can identify the specific antibody of silaenafil similar drug and the like common group, and illegal addition is detected for exploitation
The fluorescence quantum immune chromatography method of silaenafil similar drug lays the foundation.
Invention content
Based on this, the object of the present invention is to provide a kind of ternary system Immune competition methods to detect silaenafil similar drug
Quantum dot immune chromatography detection card and detection method, have increase detection signal strength and stability, improve detection sensitivity
The advantages of.
The purpose of the present invention is what is be achieved through the following technical solutions：
The quantum dot immune chromatography detection that ternary system Immune competition method detects silaenafil similar drug blocks, including liner plate,
And it is adhered to successively on liner plate and the detection antigen release pad, quantum dot probe release pad of overlapping portions between adjacent, chromatography
Film and water absorption pad；The detection antigen release pad includes that the detection made of oralbumin and the coupling of silaenafil similar drug is anti-
It is former；The quantum dot probe release pad includes to be marked with the quantum dot probe of anti-oralbumin antibody；The chromatographic film is equipped with
Detection line and nature controlling line, the detection line fix anti-silaenafil similar drug antibody, and it is anti-that the nature controlling line fixes oralbumin
It is former.
Compared to traditional " binary system Immune competition method ", detection card of the invention is utilized that " ternary system is immunized competing
Strive method " principle increases except " detection antigen " and " detection antibody " the two immune detection elements for detection antigen load
" labelled antibody " of body protein.The present invention is covalently attached detection substance (silaenafil class on carrier protein (oralbumin)
Drug) synthesis detection antigen, labelled antibody (anti-oralbumin antibody) is marked on quantum dot, by detection antibody (anti-west
That non-similar drug antibody of ground) it is fixed in chromatographic film, to detect the detection substance being coupled on antibody capture detection antigen, label
Antibody combines the carrier protein being coupled on detection antigen, forms double-antibody sandwich immune complex, generates fluorescent assay signal.When
The detection substance that detection antibody is dissociated combines, and fluorescence intensity will be suppressed, and inhibits detection signal to generate.
Specifically, dripping to sample solution in detection antigen release pad when 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 for the quantum dot probe label being carried
Albumin (OVA) antibody, detection antigen and the anti-silaenafil similar drug antibody in detection line form the immune knot of double-antibody sandwich
Object is closed, detection line is made to generate fluorescence signal；Be carried quantum dot probe label anti ova antibody with it is fixed on nature controlling line
OVA antigen bindings accumulate on nature controlling line and form fluorescence signal.Reach one when there is free silaenafil similar drug in sample
Determine concentration, immune response will be by competition blocking in detection line, and fluorescence signal is suppressed；And the fluorescence signal on nature controlling line, no
It is influenced by silaenafil similar drug concentration.
The relevant report that silaenafil similar drug is detected currently with fluorescence quantum immune chromatography method is less, the present invention
Work is detected for food and drug safety, and new tool is provided.Quantum dot is to be collected at three diameter of Spherical Volume by 200-10000 atoms
Nano semiconductor crystal, spherical nucleus diameter 2-8nm is spherical brilliant in order to increase the water solubility and biocompatibility of quantum dot
Core outer layer would generally be coated by organic molecule and introduce functional group, and diameter can be increased to 15-30nm, have good colloidal property
And kinetic characteristics, it is suitable as the Nanoparticle labeling material of Biological Detection.It is micro- with the nanometer of traditional organic fluorescent dye filling
Ball is compared, and quantum dot has width and in continuously distributed excitation spectrum, narrow and symmetrical emission peak；Stability is strong, anti-light Bleachability
By force；Light ability transformation efficiency height, brightness are that the decades of times of organic dyestuff is even more.
Compared with the existing technology, detection fixture of the invention has the advantage that：1, it is uniformly marked with anti-carrier protein antibodies
Quantum dot generate detection signal, only need centralized system for it is a kind of be used as fluorescence probe, be conducive to detect work scale and mark
Standardization；2, 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；3, steric hindrance smaller when combining is immunized, improves detection signal strength and sensitivity；4, 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, the PBS solution of sucrose are obtained.Preferably, use thickness for the glass fibre element film of 0.85mm fully absorb containing
The PBS solution of 10 μ g/mL detections antigens, 50 μ g/mL surfactants, 30mg/mL mannitol, 50mg/mL sucrose.Wherein, sweet
Dew alcohol is as freeze-drying holder for ensuring that detecting antigen in detection process dissolves rapidly；Sucrose is for adjusting detection solution viscosity control
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 silaenafil similar drug covalent coupling, can detect antibody on envelope
In conjunction with, and can be combined 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, the PBS solution of glycine are obtained.Preferably, use thickness for the artificial cellulose of 0.34mm
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 holder
Probe dissolves rapidly；Polyethylene glycol, sucrose are for adjusting detection solution viscosity control chromatography development rate；Glycine is for eliminating
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 OVA antigens for capturing the detection antigen in detection line and being fixed on nature controlling line, generate fluorescent assay signal respectively.
Further, the chromatographic film uses nitrocellulose filter, and the detection line is by containing anti-silaenafil similar drug
The PBS solution of antibody and sucrose is sprayed on nitrocellulose filter and is made.Preferably, the anti-silaenafil classes of 0.3mg/mL will be contained
The 0.05M PBS solutions (pH 7.4) of drug antibody and 10mg/mL sucrose are sprayed on nitric acid fibre with the amount of 1.0~4.0 μ g/cm
On the plain film of dimension.The basis that detection line fluorescence signal is formed is the immune anti-of silaenafil similar drug coupled by antibody and detection antigen
It answers, the silaenafil similar drug Competitive assays to dissociate in product can be detected.
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 silaenafil similar drug inspection product with the immune response of Quality Control OVA antigens, the silaenafil class dissociated in product will not be detected
Drug Competitive assays.
The quantum dot immune that ternary system Immune competition method detects silaenafil similar drug chromatographs detection method, including following
Step：
S1：Prepare detection antigen, quantum dot probe, detection line and nature controlling line；The detection antigen by oralbumin and
Silaenafil similar drug is coupled, and the quantum dot probe is marked with anti-oralbumin antibody, and the detection line is fixed anti-
Silaenafil similar drug 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 silaenafil similar drug.
Silaenafil similar drug is micromolecular compound, and the immune detection principle of conventional detection micromolecular compound is " inspection
" the binary system Immune competition method " that survey antigen " is formed with " detection antibody ", for this method in the lateral immunochromatography of quantum dot
In deficiency, the present invention on the basis of " detection antigen " is with " detection antibody ", " labelled antibody " formation is added, and " exempt from by ternary system
Epidemic disease competition law ", and combination of each immune response element in tomographic system is also adjusted, capture antibody is fixed on inspection
Survey line, labelled antibody composition fluorescence probe, forms with detection antigen (OVA- silaenafils similar drug) " bridging " over the qds
" double-antibody sandwich " immune complex, fluorescence probe accumulate to form detection signal, increase the signal strength and stability of detection,
Detection sensitivity is improved, when the detection substance that detection antibody is dissociated combines, fluorescence intensity will be suppressed, and be inhibited to generate
Detect signal.It is 2.0ng/ to the minimum detectability degree of silaenafil similar drug in sample solution using the detection method of the present invention
ML, the minimum detectability degree to illegal addition silaenafil similar drug and the like in health products, Chinese patent drug is 2.0 μ g/kg,
And it can complete quickly to detect in 5 minutes.
Further, in step S2, judged by the following method：Detection line and nature controlling line all show fluorescence signal,
Conclude that result is feminine gender, silaenafil similar drug is free of in sample；Detection line does not show that fluorescence signal, nature controlling line show fluorescence letter
Number, conclude that result is the positive, contains silaenafil similar drug in sample.Wherein, nature controlling line be for the method for inspection effectively whether
And set, nature controlling line shows that fluorescence signal shows that method is effective, and nature controlling line does not show that fluorescence signal shows that method itself is invalid；And
Detection line generates fluorescence signal when forming double-antibody sandwich immune complex, when there is free silaenafil similar drug in sample
When, the immune response in detection line will be blocked by competition, 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 that the ternary system Immune competition method of embodiment detects the quantum dot immune chromatography detection of silaenafil similar drug
The structural schematic diagram of card.
Fig. 2 a are the schematic diagram that immune response occurs in detection line, generates fluorescence signal.
Fig. 2 b are the schematic diagram that immune response is blocked by competition, fluorescence signal disappears in detection line.
Fig. 3 is the schematic diagram that immune response occurs on nature controlling line, generates fluorescence signal.
Fig. 4 is the judgment principle of the principle that fluorescence immunoassay testing result generates and testing result.
Specific implementation mode
Referring to Fig. 1, it detects the quantum dot of silaenafil similar drug for the ternary system Immune competition method of the present embodiment
Immunochromatographydetection detection card, including PVC liner plates and be adhered to successively on PVC liner plates and between adjacent overlapping portions detection it is anti-
Former release pad, quantum dot probe release pad, nitrocellulose filter (NC films) and water absorption pad；The detection antigen release pad include by
Antigen is detected made of OVA and the coupling of silaenafil similar drug；The quantum dot probe release pad includes to be marked with anti ova antibody
Quantum dot probe；The nitrocellulose filter is equipped with detection line and nature controlling line, and the detection line fixes anti-silaenafil class medicine
Product antibody, the nature controlling line fix OVA antigens.
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 and absorbs the PBS containing 10 μ g/mL detections antigens, 50 μ g/mL Triton X-100,30mg/mL mannitol, 50mg/mL sucrose
Solution, freeze-dried back.
Specifically, the preparation method of quantum dot probe release pad is as follows：Use thickness for 85 people of Whatman of 0.34mm
Cellulose membrane is made to absorb containing 20 μ g/mL quantum dot fluorescence probes, 60 μ g/mL PEG-500,10mg/mL mannitol, 60mg/mL
The PBS solution of sucrose, 10mg/mL glycine, freeze-dried back.
Specifically, the preparation method of detection line is as follows：The anti-silaenafil similar drug specific antibodies of 0.3mg/mL will be contained
It with the 0.05M PBS solutions (pH 7.4) of 10mg/mL sucrose, is sprayed on nitrocellulose filter, is formed with the amount of 1.00 μ g/cm
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.
The specific antibody induction production method of silaenafil similar drug and the like, anti ova antibody induction generation side
Method and over the qds label anti ova antibody, the method for preparing quantum dot fluorescence probe are all made of the prior art.
The quantum dot immune chromatography detection method of the silaenafil similar drug 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
So that impurity is fully precipitated within 10 minutes, 0.2mL supernatants is taken to be added to 20mL sample buffers, fully shakes 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
So that detection antigen and quantum dot probe is discharged, and successively cross detection line and nature controlling line, according to glimmering in detection line and nature controlling line
Optical signal comes in judgement sample whether contain silaenafil similar drug.
The anti ova marked on anti-silaenafil similar drug antibody, quantum dot probe in nitrocellulose filter detection line
Antibody forms double-antibody sandwich immune conjugate, so that detection line is generated fluorescent assay signal, such as respectively with detection antigen binding
Shown in Fig. 2 a.Reach a certain concentration when there is free silaenafil similar drug in sample, immune response will be by detection line
Competition blocks, and fluorescence signal can disappear, as shown in Figure 2 b.The anti ova antibody and nitrocellulose that quantum dot probe passes through label
Fixed OVA antigen bindings on film nature controlling line accumulate on nature controlling line and form fluorescence signal, as shown in Figure 3.Nature controlling line is to examine
It is set whether method itself is effective, colour developing is effective, does not develop the color and shows that method itself is invalid.
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, conclude that result is negative (A), silaenafil class is free of in sample
Drug；Detection line does not show that fluorescence signal, nature controlling line show fluorescence signal, concludes that result is positive (B and C), contains in sample
Silaenafil similar drug, wherein the Chinese and Western a concentration of 1.0ng/mL of silaenafil similar drug in positive findings B, positive findings C ground that
Non- similar drug concentration is higher than 2.0ng/mL.
Compared with the existing technology, " the ternary system Immune competition method " that the present invention uses has the following advantages：1, it uniformly uses
The quantum dot of anti-carrier protein antibodies label generates detection signal, only needs centralized system to be used as fluorescence probe for a kind of, is conducive to examine
Survey the scale and standardization of work；2, detection antibody is fixed in chromatographic film, by the closing and drying to film, to antibody
Active protection acts on advantageously, increases Antibody stability；3, steric hindrance smaller when combining is immunized, improves detection signal strength
And sensitivity.Using the detection method of the present invention to being to the minimum detectability degree of silaenafil similar drug in sample solution
2.0ng/mL, the minimum detectability degree to illegal addition silaenafil similar drug and the like in health products, Chinese patent drug is 2.0
μ g/kg, and can complete quickly to detect in 5 minutes.
Several embodiments of the invention above described embodiment only expresses, the description thereof 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 coming for those of ordinary skill in the art
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. ternary system Immune competition method detects the quantum dot immune chromatography detection card of silaenafil similar drug, it is characterised in that：
It including liner plate and is adhered on liner plate successively and the detection antigen release pad of overlapping portions, quantum dot probe is released between adjacent
Put pad, chromatographic film and water absorption pad；The detection antigen release pad include by oralbumin and silaenafil similar drug coupling and
At detection antigen；The quantum dot probe release pad includes to be marked with the quantum dot probe of anti-oralbumin antibody；It is described
Chromatographic film is equipped with detection line and nature controlling line, and the detection line fixes anti-silaenafil similar drug antibody, and the nature controlling line fixes ovum
Pure proteantigen.
2. the quantum dot immune chromatography of ternary system Immune competition method detection silaenafil similar drug according to claim 1
Detection card, it is characterised in that：The detection antigen release pad is absorbed using glass fibre element film containing detection antigen, surface-active
Agent, mannitol, the PBS solution of sucrose are obtained.
3. the quantum dot immune chromatography of ternary system Immune competition method detection silaenafil similar drug according to claim 1
Detection card, it is characterised in that：The quantum dot probe release pad using artificial cellulose's film absorb containing quantum dot fluorescence probe,
Polyethylene glycol, mannitol, sucrose, the PBS solution of glycine are obtained.
4. the quantum dot immune chromatography of ternary system Immune competition method detection silaenafil similar drug according to claim 1
Detection card, it is characterised in that：The chromatographic film uses nitrocellulose filter, and the detection line is by containing anti-silaenafil similar drug
The PBS solution of antibody and sucrose is sprayed on nitrocellulose filter and is made.
5. the quantum dot immune chromatography of ternary system Immune competition method detection silaenafil similar drug according to claim 4
Detection card, it is characterised in that：The nature controlling line is sprayed on nitrocellulose filter by the PBS solution containing oralbumin and sucrose
It is upper to be made.
6. the quantum dot immune that ternary system Immune competition method detects silaenafil similar drug chromatographs detection method, feature exists
In：Include the following steps：
S1：Prepare detection antigen, quantum dot probe, detection line and nature controlling line；The detection antigen is by oralbumin and west ground
That non-similar drug is coupled, and the quantum dot probe is marked with anti-oralbumin antibody, and the detection line fixes anti-west ground
That non-similar drug antibody, 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 silaenafil similar drug.
7. the quantum dot immune of silaenafil similar drug according to claim 6 chromatographs detection method, it is characterised in that：Step
In rapid S2, judged by the following method：Detection line and nature controlling line all show fluorescence signal, conclude that result is feminine gender, sample
In be free of silaenafil similar drug；Detection line does not show that fluorescence signal, nature controlling line show fluorescence signal, concludes that result is the positive,
Contain silaenafil similar drug in sample.
8. the quantum dot immune of silaenafil similar drug according to claim 6 chromatographs detection method, it is characterised in that：Institute
Quantum dot probe is stated under 365nm light source activations, generates the transmitting light of 630nm.
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Cited By (1)
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| CN111562384A (en) * | 2020-04-20 | 2020-08-21 | 韶关学院 | Bifunctional antigen-guided antibody array detection card for sildenafil and tadalafil |
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