CN110058012A - The fluorescence immunoassay test paper group and its preparation and application of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine - Google Patents
The fluorescence immunoassay test paper group and its preparation and application of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine Download PDFInfo
- Publication number
- CN110058012A CN110058012A CN201910275940.4A CN201910275940A CN110058012A CN 110058012 A CN110058012 A CN 110058012A CN 201910275940 A CN201910275940 A CN 201910275940A CN 110058012 A CN110058012 A CN 110058012A
- Authority
- CN
- China
- Prior art keywords
- test paper
- ketamine
- quantitative
- cocaine
- diacetylmorphine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
Abstract
The present invention provides a kind of synchronous fluorescence immunoassay test paper group and its preparation and application for detecting diacetylmorphine, crystal methamphetamine, ketamine and cocaine, it is related to biomedicine technical field, fluorescence immunoassay test paper group of the present invention, including qualitative test paper and quantitative test paper；The sample pad of qualitative test paper is respectively arranged with the fluorescent microsphere labeled monoclonal antibody area of independent four kinds of drugs, and there are four types of the mixtures of drugs for detection line spraying；The polyclonal antibody that the sample pad setting of quantitative test paper is obtained there are four types of drugs mixed immunity then sprays four kinds of drugs antigens as four quantitative detection lines on NC film respectively.Fluorescence immunoassay test paper group provided by the invention can measure four kinds of drugs simultaneously, and disposably realize qualitative and quantitative detection needs after combining qualitative test paper and quantitative test paper, time saving and energy saving, be of great significance to the on-site test of drugs law enforcement.
Description
Technical field
The present invention relates to biomedicine technical fields, more particularly to synchronous detection diacetylmorphine, crystal methamphetamine, chlorine
The fluorescence immunoassay test paper group and its preparation and application of amine ketone and cocaine.
Background technique
In recent years, the abuse of drugs has become a problem of whole world concern.Drugs are not only easily addicted, and
The physical and mental health for endangering people causes irremediable influence to the peacefulness of society.So in this illegal behavior,
We must be prevented with most fast speed, in time rescued drug addict from the environment of drug abuse, created fine
Society.It therefore, being capable of more efficiently redemption drug addict if law enfrocement official can quickly detect drugs at the scene.
Heroin is one kind of drugs such as morphine, morphine i.e. our usually described opium.It is semi-synthetic drugs, tool
There is very strong drug dependence.Heroin can not only cause very big harm to human body, and relapse rate is high, once it is stained with
Dye, almost without that may give up.It regards as level-one control drugs, and the main poison that China monitors, bans by the United Nations
One of product.
Methamphetamine is a kind of novel drug also known as meth, and main component is crystal methamphetamine, due to its shape
Shape is similar to ice, therefore is named as methamphetamine.The medicine mainly can result in human body and excitedly symptom occurs, be mainly shown as essence
Refreshing excited, sexual hyperesthesia, and along with violent tenet.Methamphetamine can reduce the flowing velocity of brain blood, lead to the dead of brain cell
It dies, makes brain by serious injury.Medication can generate very strong dependence when excessive.
Ketamine, also known as Ketamine are the derivatives of phenanthroline (PCP).It will lead to human body after taking and illusion, fortune occur
Dynamic dysfunction, is easy to produce schizophrenia, is a kind of very strong excitant.After sucking Ketamine, drug addict can constantly into
Cardiopulmonary and nervous system are caused very serious damage by row twisting, and sucking excessively can be lethal, have certain psychological dependence
Property.
Cocaine is to extract to use as anaesthetic from coca leaf earliest, but due to its strong toxicity, and can
The nervous centralis of people is set to generate excitation, the ranks for being classified as drugs have been expressly provided in countries in the world.Cocaine is to human body
Digestive system, immune system, cardiovascular system and urogenital system all have damage, can also result in necrosis of liver cells.
It is easily addicted after largely sucking, and relapse rate is more than 90%, is difficult to give up.
Infra-red sepectrometry (FTIR), gas chromatography (GC), liquid phase color are mainly used to the detection of these four drugs at present
Spectrometry (HPLC), capillary electrophoresis, gas chromatography-mass spectrometry (GC-MS) and Liquid Chromatography-Mass Spectrometry
(HPLC-MS) etc., although the comparison of these methods detection is accurate, it is longer to detect the consumed time, and to experiment people
Member and equipment have very high requirement, can not be satisfied with the on-site test of law enfrocement official outside.
Summary of the invention
The present invention provides to overcome the defect that the existing illicit drugs inspection method time is long, needs to carry out laboratory testing
A kind of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and fluorescence immunoassay test paper group of cocaine and preparation method thereof
And application method, qualitative and quantitative detection quickly can be carried out to sample to be tested, meet the needs of law enfrocement official's on-site test.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides the fluorescence of a kind of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine to exempt from
Epidemic disease test paper group, including qualitative test paper and quantitative test paper；
The qualitative test paper and quantitative test paper successively include sample pad, sample bonding pad, NC film and water suction from the bottom to top
Paper；
The sample bonding pad of the qualitative test paper includes the diacetylmorphine monoclonal antibody of independent fluorescent microsphere label
Area, the crystal methamphetamine monoclonal antibody area of fluorescent microsphere label, fluorescent microsphere label ketamine monoclonal antibody area and glimmering
The cocaine monoclonal antibody area of light microballoon label；
A qualitative detection line and a nature controlling line, the qualitative detection line are provided on the NC film of the qualitative test paper
For the mixture of diacetylmorphine, crystal methamphetamine, ketamine and cocaine；
The sample bonding pad of the quantitative test paper includes the glimmering of diacetylmorphine, crystal methamphetamine, ketamine and cocaine
Light microballoon marks polyclonal antibody；
Four quantitative detection lines and a nature controlling line, the quantitative detection line are provided on the NC film of the quantitative test paper
It is quantitatively examined including diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection line and cocaine
Survey line is respectively coated with diacetylmorphine, crystal methamphetamine, ketamine or cocaine；
The diacetylmorphine, crystal methamphetamine, ketamine and cocaine fluorescent microsphere labeled monoclonal antibody in,
The fluorescent microsphere of label is different.
Preferably, it is micro- to be selected from rare-earth fluorescent microballoon, time-resolved fluorescence microballoon, silica fluorescent for the fluorescent microsphere
One of ball and monodisperse fluorescent microsphere are a variety of.
Preferably, in the qualitative test paper and quantitative test paper, sample pad is lined with the overlapping of 1/3 length, sample in conjunction with sample
There is the overlapping of 1/3 length in one end and NC film that product bonding pad is not overlapped with sample pad, NC film do not overlapped with sample bonding pad one
There is the overlapping of 1/3 length at end with blotting paper.
Preferably, the nature controlling line is IgG polyclonal antibody.
It preferably, successively include the diacetylmorphine list of fluorescent microsphere label in the qualitative test paper, on sample bonding pad
Clonal antibody area, the crystal methamphetamine monoclonal antibody area of fluorescent microsphere label, the ketamine monoclonal of fluorescent microsphere label are anti-
The cocaine monoclonal antibody area in body area and fluorescent microsphere label.
Preferably, in the quantitative test paper, quantitative detection line is followed successively by diacetylmorphine quantitative detection line, methylbenzene third
Amine quantitative detection line, ketamine quantitative detection line and cocaine quantitative detection line.
Preferably, the fluorescent microsphere label of the diacetylmorphine, crystal methamphetamine, ketamine and cocaine is polyclonal
Antibody is the Anti-TNF-α obtained using diacetylmorphine, crystal methamphetamine, ketamine and cocaine as antigen mixed immunity
Body is marked through fluorescent microsphere.
The present invention also provides the preparation methods of fluorescence immunoassay test paper group described in above-mentioned technical proposal, comprising the following steps:
S1, diacetylmorphine monoclonal antibody area, the fluorescent microsphere mark that independent fluorescent microsphere marks are sprayed to bonding pad
What the crystal methamphetamine monoclonal antibody area of note, the ketamine monoclonal antibody area of fluorescent microsphere label and fluorescent microsphere marked
Cocaine monoclonal antibody area, obtains the bonding pad of qualitative test paper；Diacetylmorphine, crystal methamphetamine, chlorine are sprayed to bonding pad
The fluorescent microsphere of amine ketone and cocaine marks polyclonal antibody, obtains the bonding pad of quantitative test paper；
S2, to NC film spraying diacetylmorphine, crystal methamphetamine, ketamine and cocaine mixture as detection line,
IgG polyclonal antibody is sprayed as nature controlling line, obtains the NC film of qualitative test paper；Diacetylmorphine, methyl are sprayed respectively to NC film
Amphetamine, ketamine and cocaine are fixed as diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine
Detection line and cocaine quantitative detection line are measured, IgG polyclonal antibody is sprayed as nature controlling line, obtains the NC film of quantitative test paper；
S3, from bottom to top successively by sample pad, the bonding pad of qualitative test paper, the NC film of qualitative test paper and blotting paper group
Dress, obtains qualitative test paper；From bottom to top successively by sample pad, the bonding pad of quantitative test paper, the NC film of quantitative test paper and water suction
Paper assembling, obtains quantitative test paper；
S4, qualitative test paper and quantitative test paper are assembled on same sample plate, obtain fluorescence immunoassay test paper group.
Preferably, in the quantitative test paper, the spacing between each quantitative detection line is 0.8~2mm；The quantitative test paper
Quantitative detection line and nature controlling line spacing be not less than 8mm.
The present invention also provides a kind of synchronous detection diacetylmorphines, four kinds of crystal methamphetamine, ketamine and cocaine poison
The fluorescence immunoassay method of product, includes the following steps:
(1) sample to be tested is added drop-wise to the examination of fluorescence immunoassay described in technical solution described in claim 1~7 any one
The sample pad of qualitative test paper in paper group, detects the fluorescence intensity of nature controlling line and detection line after incubation, while blank control is arranged
Group；
If the fluorescence signal of qualitative test paper detection line is lower than blank control and nature controlling line has fluorescence signal, judge to be measured
Contain one of diacetylmorphine, crystal methamphetamine, ketamine and cocaine or a variety of in sample；
If the fluorescence signal intensity of qualitative test paper detection line is identical as blank control and nature controlling line has fluorescence signal, sentence
Sample to be tested break without diacetylmorphine, crystal methamphetamine, ketamine and cocaine；
(2) concentration quantitative inspection corresponding to its of diacetylmorphine, crystal methamphetamine, ketamine or cocaine is drawn respectively
The standard curve of the fluorescence intensity of survey line；
(3) by judgement in step (1) containing a kind of or more in diacetylmorphine, crystal methamphetamine, ketamine and cocaine
The sample to be tested of kind is added drop-wise to the sample pad of the quantitative test paper, and quantitative detection line is detected after incubation and the fluorescence of nature controlling line is strong
Degree calculates the one or more of diacetylmorphine in sample, crystal methamphetamine, ketamine and cocaine according to standard curve
Concentration.
Compared with prior art, beneficial effects of the present invention:
The present invention provides the fluorescence of a kind of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine to exempt from
Epidemic disease test paper group, including qualitative test paper and quantitative test paper；The sample pad of qualitative test paper is respectively arranged with the glimmering of independent four kinds of drugs
Light microballoon labeled monoclonal antibody area, there are four types of the mixtures of drugs for detection line spraying；The sample pad of quantitative test paper is provided with four
The polyclonal antibody that drugs mixed immunity obtains is planted, then sprays four kinds of drugs antigens on NC film respectively as four quantitative detections
Line.The present invention uses the principle of competition law: in the detection of qualitative test paper, four kinds of drugs to be measured will if it exists in sample
The monoclonal antibody of the fluorescent microsphere label on bonding pad corresponding to its carries out specific reaction, generated conjugate respectively
By qualitative test paper detection line when will not again with antigen binding, cause detection line fluorescence intensity reduce or disappear, thus
It determines in sample to be tested and whether contains four kinds of drugs to be measured；In the detection of quantitative test paper, it is detected as containing by qualitative test paper
There is the sample of drugs to be measured to continue quantitative detection, the drugs in sample to be tested pass through micro- with fluorescence therein when bonding pad
Ball label polyclonal antibody is specifically bound, and conjugate successively passes through four quantitative detection lines and wherein when passing through NC film
Drugs combine, after drawing the fluorescence intensity of each quantitative detection line and the standard curve of known sample concentration, can calculate
Four kinds of drugs concentration to be measured in sample to be tested.
Fluorescence immunoassay test paper group provided by the invention can measure four kinds of drugs simultaneously, and in conjunction with qualitative test paper and quantitatively
Qualitative and quantitative detection needs are disposably realized after test paper, it is time saving and energy saving, there is great meaning to the on-site test of drugs law enforcement
Justice.
It is detected using fluorescence immunoassay test paper group provided by the invention, high sensitivity (can reach 100ng/mL), specifically
Property it is strong, accuracy height (reaching 99% or more).It is low to professional knowledge and operation level requirement, drugs law enforcement field preferably
The demand of detection is of great significance to the quick primary dcreening operation of addicts.
Detailed description of the invention
Fig. 1 is the fluorescence of four kinds of synchronous quantitative detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine drugs
The front elevation of immune test paper group；
Fig. 2 is the qualitative test paper side view of fluorescence immunoassay test paper；
Fig. 3 is the quantitative test paper side view of fluorescence immunoassay test paper；
Wherein, left side is qualitative test paper, and right side is quantitative test paper；1-1 is the sample pad of qualitative test paper, and 1-2 is quantitative examination
The sample pad of paper, 2-1 are the heroin monoclonal antibody bonding pad area of fluorescent microsphere label, and 2-2 is what fluorescent microsphere marked
Methamphetamine monoclonal sample bonding pad area, 2-3 be fluorescent microsphere label ketamine monoclonal antibody bonding pad area, 2-4 be for
The cocaine monoclonal antibody bonding pad area of fluorescent microsphere label, 2-5 are the bonding pad of quantitative test paper；3-1 is qualitative test paper NC
Film, 3-2 are quantitative test paper NC film, and 4-1 is qualitative test paper nature controlling line, and 4-2 is quantitative test paper nature controlling line；5-1 is qualitative test paper
Detection line, 5-2 are the heroin quantitative detection line of quantitative test paper, and 5-3 is methamphetamine quantitative detection line, and 5-4 is that ketamine is quantitatively examined
Survey line, 5-5 are cocaine quantitative detection line, and 6-1 is qualitative test paper blotting paper, and 6-2 is quantitative test paper blotting paper, and 7-1 is qualitative
Test paper PVC bottom plate, 7-2 are quantitative test paper PVC bottom plate, and 8 be sample panel.
Specific embodiment
The present invention provides the fluorescence of a kind of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine to exempt from
Epidemic disease test paper group, including qualitative test paper and quantitative test paper；
The qualitative test paper and quantitative test paper successively include sample pad, sample bonding pad, NC film and water suction from the bottom to top
Paper；
The sample bonding pad of the qualitative test paper includes the diacetylmorphine monoclonal antibody of independent fluorescent microsphere label
Area, the crystal methamphetamine monoclonal antibody area of fluorescent microsphere label, fluorescent microsphere label ketamine monoclonal antibody area and glimmering
The cocaine monoclonal antibody area of light microballoon label；
A qualitative detection line and a nature controlling line, the qualitative detection line are provided on the NC film of the qualitative test paper
For the mixture of diacetylmorphine, crystal methamphetamine, ketamine and cocaine；
The sample bonding pad of the quantitative test paper includes the glimmering of diacetylmorphine, crystal methamphetamine, ketamine and cocaine
Light microballoon marks polyclonal antibody；
Four quantitative detection lines and a nature controlling line, the quantitative detection line are provided on the NC film of the quantitative test paper
It is quantitatively examined including diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection line and cocaine
Survey line is respectively coated with diacetylmorphine, crystal methamphetamine, ketamine or cocaine；
The diacetylmorphine, crystal methamphetamine, ketamine and cocaine fluorescent microsphere labeled monoclonal antibody in,
The fluorescent microsphere of label is different.
In qualitative test paper of the present invention and quantitative test paper, in order to make sample adequately absorb and react,
Preferably have the overlapping of 1/3 length between the sample pad and sample bonding pad, sample bonding pad do not overlapped with sample pad one
Preferably there is the overlapping of 1/3 length at end with NC film, and one end and blotting paper that NC film is not overlapped with sample bonding pad preferably have 1/
3 length overlap.
In the present invention, the material of the sample pad is preferably glass fibre membrane.Sample pad is for being added dropwise sample.In order to
Improve detection sensitivity, reduce sample variation, it is currently preferred sample pad is soaked in sample pad preprocessing solution after
It is dry；By mass percentage, the sample pad preprocessing solution is spat including 1~5%BSA, 1~4% sucrose, 0.5~2%
The water of warm -20,0.01~0.05% Sodium azide, 0.1~0.5% PVPK-30 and surplus；More preferably include 3%BSA,
2.5% sucrose, 1% Tween-20,0.02% Sodium azide, 0.3%PVPK-30 and surplus water.In the present invention, the immersion
Time is preferably 20~50min, more preferably 30~40min.
In the present invention, the material of the sample bonding pad is preferably glass fibre.Sample bonding pad is coated with to be measured four
The monoclonal antibody or polyclonal antibody of kind of drugs, so as on sample to be tested and sample bonding pad monoclonal antibody or more grams
Grand antibody combines.It is preferred successively (from sample pad to NC on the sample bonding pad in qualitative test paper of the present invention
Film direction) spray the diacetylmorphine monoclonal antibody of fluorescent microsphere label, the crystal methamphetamine monoclonal of fluorescent microsphere label
The cocaine monoclonal antibody of antibody, the ketamine monoclonal antibody of fluorescent microsphere label and fluorescent microsphere label.
In the present invention, the fluorescent microsphere for marking the monoclonal antibody include but is not limited to rare-earth fluorescent microballoon,
Four kinds in time-resolved fluorescence microballoon, silica fluorescent microballoon and monodisperse fluorescent microsphere.In the present invention, for marking
The fluorescent microsphere for remembering the polyclonal antibody of diacetylmorphine, crystal methamphetamine, ketamine and cocaine includes but is not limited to rare earth
Fluorescent microsphere, time-resolved fluorescence microballoon, in silica fluorescent microballoon and monodisperse fluorescent microsphere any one or it is more
Kind.In the present invention, when carrying out fluorescent microsphere label, preferably fluorescent microsphere is activated using EDC and NHS solution, institute
Stating the preferred concentration of EDC and NHS solution is 5mg/mL.
In the present invention, the sample bonding pad is before spraying monoclonal antibody or polyclonal antibody, preferably with buffering
Liquid is pre-processed；The buffer includes 15~30% sucrose of quality volume fraction, the small ox blood of quality volume fraction 1~6%
The Tris-HCl buffer of pure albumen and 0.02~0.1M；It more preferably include 20% sucrose of quality volume fraction, mass body
The Tris-HCl buffer of 2% balf serum albumin of fraction and 0.05M.In the present invention, the buffer is pretreated
Time is preferably 1.5~4h, more preferably 2h, dry after being pre-processed with buffer.
In the present invention, the methyl that the diacetylmorphine monoclonal antibody of the fluorescent microsphere label, fluorescent microsphere mark
The cocaine monoclonal of amphetamine monoclonal antibody, the ketamine monoclonal antibody of fluorescent microsphere label or fluorescent microsphere label
Antibody sources are prepared in commercial goods or voluntarily.It is described when voluntarily preparing, it is currently preferred with diacetylmorphine, first
Base amphetamine, ketamine or cocaine are that animal is immunized in antigen, obtain diacetylmorphine, crystal methamphetamine, ketamine
Or it is marked again with fluorescent microsphere after the monoclonal antibody of cocaine.
In the present invention, the fluorescent marker of the diacetylmorphine, crystal methamphetamine, ketamine and cocaine is polyclonal
Antibody can be specifically bound with four kinds of diacetylmorphine, crystal methamphetamine, ketamine and cocaine drugs, can be derived from
Commercial goods voluntarily prepare.In the present invention, the fluorescence of the diacetylmorphine, crystal methamphetamine, ketamine and cocaine
Microballoon label polyclonal antibody preparation be preferably: using diacetylmorphine, crystal methamphetamine, ketamine and cocaine as
Antigen, polyclonal antibody obtained from the same animal of co-immunization after mixing, then marked with fluorescent microsphere.
It in the present invention, include a detection line and a nature controlling line, the inspection on NC film in the qualitative test paper
Survey line is the mixture of diacetylmorphine, crystal methamphetamine, ketamine and cocaine, is used for and " sample-fluorescent microsphere Dan Ke
The being at war with property of conjugate of grand antibody " combines, so that the conjugate is fixed on detection line position, if not containing in sample
Four kinds of drugs to be measured, then four in detection line kind drugs can mark monoclonal anti-to the corresponding fluorescent microsphere in sample pad
Body combines, so that there are higher fluorescence intensities for detection line；Conversely, containing in the case where four kinds of drugs to be measured in sample
The fluorescence intensity of detection line can reduce accordingly.Therefore, the present invention can be to sample by the fluorescence intensity size of detection line
In whether containing four kinds of drugs to be measured carry out it is qualitative, it is simple and convenient, quickly can determine and exclude drug addict.Specific
It detects under environment, it can be by blank sample as compareing, thus the detection line fluorescence intensity change in more qualitative test paper.
In the present invention, described including four quantitative detection lines and a nature controlling line on NC film in the quantitative test paper
Four quantitative detection lines be respectively that diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine are quantitatively examined
Survey line and cocaine quantitative detection line, are respectively coated with diacetylmorphine, crystal methamphetamine, ketamine or cocaine.Described
Four quantitative detection lines are the antigen of four kinds of drugs, are respectively independently arranged, dense to distinguish specifically which kind of drugs and drugs
Degree.When quantitative detection, four detection lines difference can carry out specificity with " sample-fluorescent marker polyclonal antibody "
In conjunction with, when " sample-fluorescent microsphere mark polyclonal antibody " passes through each quantitative detection line, then this quantitative detection line
On drugs antigen specifically bound with polyclonal antibody therein, if not containing this kind of drugs in sample, this is fixed
It measures in conjunction with the antigen in detection line all marks polyclonal antibody with fluorescent microsphere, fluorescence intensity highest at this time；If containing in sample
There is this kind of virus, then the fluorescent microsphere label polyclonal antibody that the antigen on this quantitative detection line can combine is reduced, thus
Fluorescence intensity reduces.Therefore, pass through the standard curve of fluorescence intensity size in the drugs of known concentration and detection line, Ji Keji
The concentration for calculating contained this kind of drugs in corresponding sample, realizes quantitative detection.
In the present invention, four quantitative detections put in order it is preferred successively (from sample bonding pad to the film side NC
To) it is that diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection line and cocaine are quantitative
Detection line.
In the present invention, the nature controlling line is IgG polyclonal antibody.In qualitative detection and quantitative detection, not with inspection
Survey line combine part monoclonal antibody or polyclonal antibody pass through nature controlling line when, can in conjunction with IgG polyclonal antibody, from
And generate fluorescence intensity.The purpose that nature controlling line is arranged in the present invention is that test strips itself fail in order to prevent, when nature controlling line unstressed configuration
When intensity, then testing result is invalid.
In the present invention, it from sample bonding pad to NC film direction, is arranged successively as detection line, nature controlling line.
In the present invention, the test paper group further includes bottom plate, for fixing qualitative test paper and quantitative test paper together.?
In the present invention, the bottom plate is do not absorb water material, preferably PVC.
The present invention also provides the preparation methods of fluorescence immunoassay test paper group described in above-mentioned technical proposal, comprising the following steps:
S1, diacetylmorphine monoclonal antibody area, the fluorescent microsphere mark that independent fluorescent microsphere marks are sprayed to bonding pad
What the crystal methamphetamine monoclonal antibody area of note, the ketamine monoclonal antibody area of fluorescent microsphere label and fluorescent microsphere marked
Cocaine monoclonal antibody area, obtains the bonding pad of qualitative test paper；Diacetylmorphine, crystal methamphetamine, chlorine are sprayed to bonding pad
The fluorescent microsphere of amine ketone and cocaine marks polyclonal antibody, obtains the bonding pad of quantitative test paper；
S2, to NC film spraying diacetylmorphine, crystal methamphetamine, ketamine and cocaine mixture as detection line,
IgG polyclonal antibody is sprayed as nature controlling line, obtains the NC film of qualitative test paper；Diacetylmorphine, methyl are sprayed respectively to NC film
Amphetamine, ketamine and cocaine are fixed as diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine
Detection line and cocaine quantitative detection line are measured, IgG polyclonal antibody is sprayed as nature controlling line, obtains the NC film of quantitative test paper；
S3, from bottom to top successively by sample pad, the bonding pad of qualitative test paper, the NC film of qualitative test paper and blotting paper group
Dress, obtains qualitative test paper；From bottom to top successively by sample pad, the bonding pad of quantitative test paper, the NC film of quantitative test paper and water suction
Paper assembling, obtains quantitative test paper；
S4, qualitative test paper and quantitative test paper are assembled on same sample plate, obtain fluorescence immunoassay test paper group.
In the present invention, in order to further increase the accuracy of quantitative detection, reduce error, in the quantitative test paper, respectively
Spacing between quantitative detection line is preferably 0.8~2mm, more preferably 1mm；The quantitative detection line and matter of the quantitative test paper
The spacing of control line is preferably not less than 8mm.
The present invention also provides a kind of synchronous detection diacetylmorphines, four kinds of crystal methamphetamine, ketamine and cocaine poison
The fluorescence immunoassay method of product, includes the following steps:
(1) sample to be tested is added drop-wise to the sample of the qualitative test paper in fluorescence immunoassay test paper group described in preceding solution
Product pad, detects the fluorescence intensity of nature controlling line and detection line after incubation, while blank control group is arranged；
If the fluorescence signal of qualitative test paper detection line is lower than blank control and nature controlling line has fluorescence signal, judge to be measured
Contain one of diacetylmorphine, crystal methamphetamine, ketamine and cocaine or a variety of in sample；
If the fluorescence signal intensity of qualitative test paper detection line is identical as blank control and nature controlling line has fluorescence signal, sentence
Sample to be tested break without diacetylmorphine, crystal methamphetamine, ketamine and cocaine；
(2) concentration quantitative inspection corresponding to its of diacetylmorphine, crystal methamphetamine, ketamine or cocaine is drawn respectively
The standard curve of the fluorescence intensity of survey line；
(3) by judgement in step (1) containing a kind of or more in diacetylmorphine, crystal methamphetamine, ketamine and cocaine
The sample to be tested of kind is added drop-wise to the sample pad of the quantitative test paper, and quantitative detection line is detected after incubation and the fluorescence of nature controlling line is strong
Degree calculates the one or more of diacetylmorphine in sample, crystal methamphetamine, ketamine and cocaine according to standard curve
Concentration.
In the present invention, for the sample to be tested amount of taking of detection preferably at 80~200 μ L/ times.In the present invention,
Sample to be tested can be solid or liquid, to be detected again after water or organic solvent dissolution if sample to be tested is solid, example
The flour of drugs to be measured may be such as mixed with；Sample to be tested can be body fluid, the beverage etc. that may contain drugs to be measured.
The high sensitivity that is detected using test paper group of the present invention, accuracy are high, easy to operate, are suitable for scene
Detection.
Technical solution provided by the invention is described in detail below with reference to embodiment, but they cannot be managed
Solution is limiting the scope of the present invention.
Embodiment 1
It is of the present invention to be used for while detecting diacetylmorphine, crystal methamphetamine, ketamine and cocaine four kinds of drugs
Test paper group includes that qualitative test paper and quantitative test paper are constituted.The structure of qualitative test paper and quantitative test paper is followed successively by sample from bottom to top
Pad, bonding pad, NC film and blotting paper.Sample pad, bonding pad, NC film and blotting paper are adhered on PVC bottom plate.Sample pad with
Bonding pad fibreglass film is constituted, and reaction film is then nitrocellulose filter, and blotting paper is strong water absorption function paper.Specific knot
Structure is as shown in Figure 1.
1, the preparation of qualitative test paper:
The sample pad of qualitative test strips is divided into independent four areas, is followed successively by the diacetylmorphine list of fluorescent microsphere label
Clonal antibody area, the crystal methamphetamine monoclonal antibody area of fluorescent microsphere label, the ketamine monoclonal of fluorescent microsphere label are anti-
The cocaine monoclonal antibody area in body area and fluorescent microsphere label；Detection line and nature controlling line, detection line are successively coated on NC film
For the mixture of diacetylmorphine, crystal methamphetamine, ketamine and cocaine.
1) preparation of monoclonal antibody
The synthetic antigen that acetylmorphine, crystal methamphetamine, ketamine or cocaine is respectively adopted passes through the side of abdomen injection
Formula is immunized BALB/c mouse, then by preparing hybridoma and carrying out monoclonal antibody in Mice Body using hybridoma
Large-scale production.The monoclonal antibody produced must need octanoic acid-ammonium sulfate precipitation method to be purified, and finally obtain two
Acetylmorphine monoclonal antibody, crystal methamphetamine monoclonal antibody, ketamine monoclonal antibody, cocaine monoclonal antibody.
2) fluorescent microsphere-antibody conjugates preparation
With PBS solution by fluorescent microsphere grain dissolution, and EDC and NHS solution is added and is activated, its compound is put
37 DEG C of incubator 1h are placed in, it is spare that precipitating is collected by centrifugation.Various monoclonal antibodies prepared by step 1), respectively and after activation
Fluorescent microsphere be combined, supernatant is removed in centrifugation after concussion shakes up, and precipitating wash at least 2 times with PBS, adds after ultrasonic treatment
Enter PBS buffer solution and places 4 DEG C of refrigerators.
3) preparation of sample pad
The material of sample pad is glass fibre membrane, and being soaked in chemical substance, (3%BSA, 2.5% sucrose, 1% are spat
Warm -20,0.02% Sodium azide and 0.3% PVPK-30) in half an hour, abundant drying for standby.To improve sensitivity, subtract
The otherness of few sample.
4) preparation of bonding pad
Bonding pad is glass fibre membrane, in advance using buffer (20% sucrose, 2% balf serum albumin,
The Tris-Hcl of 0.05%M) carry out 2h processing, handle afterwards 37 DEG C of drying for standby.By four kinds of drugs of step 2) preparation
Fluorescent microsphere-monoclonal antibody is sprayed onto respectively on the corresponding region of sample bonding pad, is subsequently placed at 37 DEG C and is incubated 2h,
It is spare after taking-up.
5) preparation of NC film
Containing a detection line and a nature controlling line on qualitative test strips NC film, wire spraying Heroine antigens, methamphetamine are detected
Antigen, ketamine antigen and cocaine antigen mixture.Nature controlling line is that sheep anti-mouse igg is mostly anti-, be will test respectively using synovial membrane instrument
Line and nature controlling line slide on NC film, and the spacing of every detection line is 1mm.Detection line is 8mm at a distance from nature controlling line.It will preparation
It is spare that good NC film places dry 1h at 37 DEG C.
2, the preparation of quantitative test paper:
The sample pad of quantitative test paper item is that the fluorescence of diacetylmorphine, crystal methamphetamine, fluorescence ketamine and cocaine is micro-
Ball marks polyclonal antibody；Four detection lines and a nature controlling line are successively coated on NC film, detection line is followed successively by diacetyl
Coffee quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection line and cocaine quantitative detection line.
1) preparation of monoclonal antibody
By the way of the synthetic antigen mixing postabdomen injection of acetylmorphine, crystal methamphetamine, ketamine or cocaine
It is immunized BALB/c mouse, then by preparing hybridoma and carrying out the big of monoclonal antibody in Mice Body using hybridoma
Large-scale production.The polyclonal antibody produced must need octanoic acid-ammonium sulfate precipitation method to be purified, and finally obtain diethyl
Acyl morphine, crystal methamphetamine, ketamine and cocaine polyclonal antibody.
2) fluorescent microsphere-antibody conjugates preparation
With PBS solution by fluorescent microsphere grain dissolution, and EDC and NHS solution is added and is activated, its compound is put
37 DEG C of incubator 1h are placed in, it is spare that precipitating is collected by centrifugation.By step 1) prepare polyclonal antibody, respectively with it is glimmering after activation
Light microballoon is combined, and supernatant is removed in centrifugation after concussion shakes up, and precipitating is washed at least 2 times with PBS, PBS is added after ultrasonic treatment
Buffer places 4 DEG C of refrigerators.
3) preparation of sample pad
The material of sample pad is glass fibre membrane, and being soaked in chemical substance, (3%BSA, 2.5% sucrose, 1% are spat
Warm -20,0.02% Sodium azide and 0.3% PVPK-30) in half an hour, abundant drying for standby.To improve sensitivity, subtract
The otherness of few sample.
4) preparation of bonding pad
Bonding pad is glass fibre membrane, in advance using buffer (20% sucrose, 2% balf serum albumin,
The Tris-Hcl of 0.05%M) carry out 2h processing, handle afterwards 37 DEG C of drying for standby.Fluorescent microsphere-prepared by step 2)
Polyclonal antibody is sprayed on sample bonding pad, is subsequently placed at 37 DEG C and is incubated 2h, spare after taking-up.
5) preparation of NC film
Containing four detection lines and a nature controlling line on qualitative test strips NC film, it is anti-that Heroine antigens, methamphetamine are sprayed respectively
Former, ketamine antigen and cocaine antigen are detected as four.Nature controlling line is that sheep anti-mouse igg is mostly anti-, respectively will using synovial membrane instrument
Detection line and nature controlling line slide on NC film, and four detection lines successively contain Heroine antigens, methamphetamine antigen, chloramines from top to bottom
Ketone antigen and cocaine antigen.The spacing of every detection line is 1mm.Heroine Detection line is 8mm at a distance from nature controlling line.It will
It is spare that the NC film prepared places dry 1h at 37 DEG C.
It is assembled, is obtained according to the sequence for being followed successively by sample pad, sample bonding pad, NC film and blotting paper from bottom to top
Quantitative test paper and qualitative test paper.Wherein, 1/3 on the downside of bonding pad is located at the lower section of sample pad, and the 1/3 of upside is located at NC film
Top is conducive to sample in this way and preferably absorbs and combine.Sample pad, bonding pad, NC film and blotting paper are on PVC bottom plate.
Test strips are cut into 6mm width size after being completed, be subsequently mounted into detection in getting stuck, then will block
Quantitative analysis is carried out in shell insertion fluorescence detector.
The formulation of 2 standard curve of embodiment
The concentration of standard items and final fluorescence intensity are done into standard curve, can thus pass through the glimmering of sample
Luminous intensity carries out quantitative analysis.
The diacetylmorphine of known concentration, crystal methamphetamine, ketamine and cocaine are diluted to following concentration system respectively
Column: 1000ng/mL, 500ng/mL, 250ng/mL, 100ng/mL, 50ng/mL, 25ng/mL, 10ng/mL, 0ng/mL.
It takes the sample of 100 each concentration of μ L to be added drop-wise in the sample pad of the quantitative test paper of the building of embodiment 1, is stood at 25 DEG C
5min；The inspection of fluorescence intensity size is carried out to the detection line and nature controlling line reacted in the test paper completed with immunofluorescence analysis instrument
It surveys.Replication 3 times, respectively using obtained fluorescence intensity signals T/C value as ordinate.It is done concentration as abscissa
Standard curve.It is written into the system of instrument to carry out quantitative analysis later.
The standard curve of diacetylmorphine is y=6E-06x2- 0.0089x+5.4502, R2It is 0.9786.
The standard curve of crystal methamphetamine is y=3E-06x2- 0.0044x+4.7449, R2It is 0.9811.
The standard curve of ketamine is y=2E-06x2- 0.0033x+3.7098, R2It is 0.9732.
The standard curve of cocaine is y=5E-06x2- 0.0077x+4.1097, R2It is 0.9908.
1, sensitivity determination
The measurement that sensitivity is carried out using the quantitative test paper of test paper group carries out fluorescence intensity from high concentration to low concentration
Observation, the as detection line when being presented negative.The fluorescence intensity of T line is better than C line when negative, so, by the detection of quantitative test paper
Line is compared with C line, to determine the detection line of 4 kinds of drugs respectively.The sensitivity for finally obtaining diacetylmorphine is 100ng/
ML, the sensitivity of crystal methamphetamine are 500ng/mL, and the sensitivity of ketamine is 500ng/mL, and the sensitivity of cocaine is
100ng/mL。
2, specific assay
The specificity of test paper can be measured by the qualitative test paper of test paper group.Fentanyl, C16H25NO2, Ah Si are taken respectively
Woods, berberine, diacetylmorphine, crystal methamphetamine, ketamine, cocaine, phenobarbital, Ofloxacin, flutters heat at orfloxacin
Breath pain is just leading to 17 kinds of drugs such as peaceful piece, Lofexidine, naloxone, clonidine and alprazolam and the specific survey of drugs progress
It is fixed.The results show that only positive findings are presented in diacetylmorphine, crystal methamphetamine, ketamine and cocaine, remaining is
Cross reaction does not occur for feminine gender.The result shows that this test paper group is to diacetylmorphine, crystal methamphetamine, ketamine and cocaine
Detection have stronger specificity.
Embodiment 3
The measurement that 50 people carry out its urine is randomly selected in certain public place of entertainment, 100 μ L urine samples is taken to be added to embodiment
In the qualitative test paper sample pad of the test paper group of 1 building, 5min is stood at 25 DEG C, qualitative examination is detected using immunofluorescence analysis instrument
The detection line and nature controlling line of paper, and as a control group with clear water.If the detection line fluorescence intensity of sample lower than control group and
Nature controlling line has fluorescence intensity, then judges in the sample containing in diacetylmorphine, crystal methamphetamine, ketamine and cocaine
It is one or more, it is denoted as the positive；Conversely, if the detection line fluorescence intensity of sample is identical as control group and nature controlling line has fluorescence strong
Degree then judges that the sample without containing diacetylmorphine, crystal methamphetamine, ketamine and cocaine, is denoted as feminine gender.
Obtain there is the urine sample of 3 people to be positive result after testing.By this 50 parts of urine specimens be sent to public security organ into
Row detection, testing result in detail is consistent with test strips testing result of the invention, it was demonstrated that the accuracy of qualitative detection of the invention
It is very high, it is able to carry out the primary dcreening operation work at scene, and to operator without the technical requirements of especially big difficulty.
Embodiment 4
The blood sample that 25 people are chosen from narcotic house carries out qualitative analysis.The above-mentioned 25 parts of blood samples of 100 μ L are taken respectively
This, is added drop-wise in the sample pad of the quantitative test paper of the building of embodiment 1 respectively, stands 5min at 25 DEG C；Fluorescence analyser is immunized
The detection for the detection line and nature controlling line progress fluorescence intensity size in test paper that reaction is completed.
It is analyzed, determines its drug species sucked by observing the situation of change of these four detection lines first, then
It is compared with data provided by narcotic house, as the result is shown unanimously.
Then carry out quantitative analysis, using fluorescence analyser by most start measurement concentration record, then carry out for
Phase one month monitoring, diagnosing, the blood for extracting smoker daily carry out the monitoring of drugs concentration.Pass through one month prison
It surveys, it is found that most of intracorporal toxin concentration of smoker is substantially reduced, and the testing result of this month is mentioned with narcotic house
The result of confession is roughly the same.Compared with test paper group provided by the invention detecting instrument used by the narcotic house, to operator's skill
Art require it is low and convenient and efficient, can the intracorporal endotoxin level of real-time detection smoker, and take phase according to the actual situation
The measure answered.
As seen from the above embodiment, test paper group provided by the invention can quickly, be completed at the same time to diacetylmorphine, methyl
The on-site test of four kinds of amphetamine, ketamine and cocaine drugs, accuracy height (being consistent with Laboratory Instruments testing result),
Quantitative detection result is accurate, simple portable, easily operated.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications
It should be regarded as protection scope of the present invention.
Claims (10)
1. the fluorescence immunoassay test paper group of a kind of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine, feature
It is, including qualitative test paper and quantitative test paper；
The qualitative test paper and quantitative test paper successively include sample pad, sample bonding pad, NC film and blotting paper from the bottom to top；
The sample bonding pad of the qualitative test paper includes the diacetylmorphine monoclonal antibody area, glimmering of independent fluorescent microsphere label
The crystal methamphetamine monoclonal antibody area of light microballoon label, the ketamine monoclonal antibody area of fluorescent microsphere label and fluorescent microsphere
The cocaine monoclonal antibody area of label；
A qualitative detection line and a nature controlling line are provided on the NC film of the qualitative test paper, the qualitative detection line is diethyl
Acyl morphine, crystal methamphetamine, ketamine and cocaine mixture；
The sample bonding pad of the quantitative test paper includes that the fluorescence of diacetylmorphine, crystal methamphetamine, ketamine and cocaine is micro-
Ball marks polyclonal antibody；
Four quantitative detection lines and a nature controlling line are provided on the NC film of the quantitative test paper, the quantitative detection line includes two
Acetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection line and cocaine quantitative detection line, respectively
From being coated with diacetylmorphine, crystal methamphetamine, ketamine or cocaine；
The diacetylmorphine, crystal methamphetamine, ketamine and cocaine fluorescent microsphere labeled monoclonal antibody in, label
Fluorescent microsphere is different.
2. fluorescence immunoassay test paper group according to claim 1, which is characterized in that it is micro- that the fluorescent microsphere is selected from rare-earth fluorescent
One of ball, time-resolved fluorescence microballoon, silica fluorescent microballoon and monodisperse fluorescent microsphere are a variety of.
3. fluorescence immunoassay test paper group according to claim 1, which is characterized in that in the qualitative test paper and quantitative test paper,
Sample pad is lined with the overlapping of 1/3 length in conjunction with sample, and there are 1/3 length in one end and NC film that sample bonding pad is not overlapped with sample pad
Degree overlaps, and there be the overlapping of 1/3 length in one end and blotting paper that NC film is not overlapped with sample bonding pad.
4. fluorescence immunoassay test paper group according to claim 1, which is characterized in that the nature controlling line is IgG polyclonal antibody.
5. fluorescence immunoassay test paper group according to claim 1, which is characterized in that in the qualitative test paper, sample bonding pad
On successively include fluorescent microsphere label diacetylmorphine monoclonal antibody area, fluorescent microsphere label crystal methamphetamine monoclonal
The cocaine monoclonal antibody area that antibody district, the ketamine monoclonal antibody area of fluorescent microsphere label and fluorescent microsphere mark.
6. fluorescence immunoassay test paper group according to claim 1 or 5, which is characterized in that in the quantitative test paper, quantitative detection
It is fixed that line is followed successively by diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection line and cocaine
Measure detection line.
7. fluorescence immunoassay test paper group according to claim 1, which is characterized in that the diacetylmorphine, crystal methamphetamine,
The fluorescent microsphere of ketamine and cocaine marks polyclonal antibody, is with diacetylmorphine, crystal methamphetamine, ketamine and Ke Ka
It is marked because of the polyclonal antibody obtained as antigen mixed immunity through fluorescent microsphere.
8. the preparation method of fluorescence immunoassay test paper group described in claim 1~7 any one, comprising the following steps:
S1, diacetylmorphine monoclonal antibody area, fluorescent microsphere that independent fluorescent microsphere marks are sprayed to bonding pad mark
The cocaine in crystal methamphetamine monoclonal antibody area, the ketamine monoclonal antibody area of fluorescent microsphere label and fluorescent microsphere label
Monoclonal antibody area obtains the bonding pad of qualitative test paper；To bonding pad spraying diacetylmorphine, crystal methamphetamine, ketamine and
The fluorescent microsphere of cocaine marks polyclonal antibody, obtains the bonding pad of quantitative test paper；
S2, the mixture of diacetylmorphine, crystal methamphetamine, ketamine and cocaine is sprayed to NC film as detection line, spraying
IgG polyclonal antibody obtains the NC film of qualitative test paper as nature controlling line；Diacetylmorphine, methylbenzene third are sprayed respectively to NC film
Amine, ketamine and cocaine are as diacetylmorphine quantitative detection line, crystal methamphetamine quantitative detection line, ketamine quantitative detection
Line and cocaine quantitative detection line spray IgG polyclonal antibody as nature controlling line, obtain the NC film of quantitative test paper；
S3, successively sample pad, the bonding pad of qualitative test paper, the NC film of qualitative test paper and blotting paper are assembled from bottom to top, is obtained
To qualitative test paper；From bottom to top successively by sample pad, the bonding pad of quantitative test paper, the NC film of quantitative test paper and blotting paper group
Dress, obtains quantitative test paper；
S4, qualitative test paper and quantitative test paper are assembled on same sample plate, obtain fluorescence immunoassay test paper group.
9. preparation method according to claim 8, which is characterized in that in the quantitative test paper, between each quantitative detection line
Spacing be 0.8~2mm；The quantitative detection line of the quantitative test paper and the spacing of nature controlling line are not less than 8mm.
10. the fluorescence immunoassay side of a kind of synchronous detection diacetylmorphine, four kinds of crystal methamphetamine, ketamine and cocaine drugs
Method includes the following steps:
(1) sample to be tested is added drop-wise in fluorescence immunoassay test paper group described in technical solution described in claim 1~7 any one
Qualitative test paper sample pad, the fluorescence intensity of nature controlling line and detection line is detected after incubation, while blank control group is set；
If the fluorescence signal of qualitative test paper detection line is lower than blank control and nature controlling line has fluorescence signal, judge in sample to be tested
Contain one of diacetylmorphine, crystal methamphetamine, ketamine and cocaine or a variety of；
If the fluorescence signal intensity of qualitative test paper detection line is identical as blank control and nature controlling line has fluorescence signal, judge to be measured
Sample is without diacetylmorphine, crystal methamphetamine, ketamine and cocaine；
(2) concentration quantitative detection line corresponding to its of diacetylmorphine, crystal methamphetamine, ketamine or cocaine is drawn respectively
The standard curve of fluorescence intensity；
(3) will judge in step (1) containing one or more in diacetylmorphine, crystal methamphetamine, ketamine and cocaine
Sample to be tested is added drop-wise to the sample pad of the quantitative test paper, and the fluorescence intensity of quantitative detection line and nature controlling line, root are detected after incubation
One or more concentration of diacetylmorphine, crystal methamphetamine, ketamine and cocaine in sample are calculated according to standard curve.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910275940.4A CN110058012B (en) | 2019-04-08 | 2019-04-08 | Fluorescent immune test paper group for synchronously detecting diacetylmorphine, methamphetamine, ketamine and cocaine and preparation and use methods thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910275940.4A CN110058012B (en) | 2019-04-08 | 2019-04-08 | Fluorescent immune test paper group for synchronously detecting diacetylmorphine, methamphetamine, ketamine and cocaine and preparation and use methods thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110058012A true CN110058012A (en) | 2019-07-26 |
| CN110058012B CN110058012B (en) | 2022-03-01 |
Family
ID=67318567
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910275940.4A Active CN110058012B (en) | 2019-04-08 | 2019-04-08 | Fluorescent immune test paper group for synchronously detecting diacetylmorphine, methamphetamine, ketamine and cocaine and preparation and use methods thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110058012B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110850095A (en) * | 2019-11-22 | 2020-02-28 | 佛山墨赛生物技术有限公司 | Drug trace detection test strip and preparation method and kit thereof |
| CN111474342A (en) * | 2020-04-20 | 2020-07-31 | 石家庄洹众生物科技有限公司 | Colloidal gold detection card for four-item joint detection of drugs and method for detecting drugs |
| CN113391062A (en) * | 2021-05-25 | 2021-09-14 | 黄淮学院 | Morphine immunofluorescence chromatography rapid detection test paper strip, preparation method and detection method thereof |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4207307A (en) * | 1977-01-21 | 1980-06-10 | Research Corporation | Simultaneous immunoassay of multiple antigens and assay for cocaine metabolites |
| US5962336A (en) * | 1997-10-20 | 1999-10-05 | Sun; Ming | Multi-test panel |
| CN101556280A (en) * | 2009-05-20 | 2009-10-14 | 江西中德生物工程有限公司 | Immunochromatography detection card for detecting fluorescent microspheres of ketamine and methyl amphetamine and preparation method thereof |
| CN202854140U (en) * | 2012-06-07 | 2013-04-03 | 北京维德维康生物技术有限公司 | Clenbuterol rapid triple detection card |
| CN103336113A (en) * | 2013-06-24 | 2013-10-02 | 苏州万木春生物技术有限公司 | Preparation method of human immunodeficiency virus (HIV) antibody detection test paper |
| CN104634967A (en) * | 2014-11-30 | 2015-05-20 | 德康润生物科技（天津）有限公司 | Method for further detecting four narcotics in saliva by employing immuno-chromatographic technology |
| CN104655837A (en) * | 2015-02-27 | 2015-05-27 | 南京微测生物科技有限公司 | Fluorescent quantitative test paper strip for simultaneously detecting algal toxins MC-LR/RR/YR and preparation method and application of fluorescent quantitative test paper strip |
| CN105277693A (en) * | 2014-08-18 | 2016-01-27 | 董俊 | Human parainfluenza virus quantum dot immunochromatography typing detection card, preparation method and applications |
| CN107942062A (en) * | 2017-11-29 | 2018-04-20 | 洛阳现代生物技术研究院有限公司 | Test card, preparation and the detection method of synchronous detection 2 toxin of ochracin, vomitoxin and T |
| CN108845151A (en) * | 2018-05-29 | 2018-11-20 | 郑州左安检测科技有限公司 | A kind of test strips and its preparation method and application method detecting crystal methamphetamine |
| CN208607233U (en) * | 2018-06-29 | 2019-03-15 | 江苏达骏生物科技有限公司 | Simultaneous quantitative detects the reagent strip and test card of quinolone, tetracycline and sulfanilamide (SN) |
| CN109490546A (en) * | 2018-12-21 | 2019-03-19 | 广东顺德工业设计研究院(广东顺德创新设计研究院) | Six detection devices of drugs and preparation method thereof |
-
2019
- 2019-04-08 CN CN201910275940.4A patent/CN110058012B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4207307A (en) * | 1977-01-21 | 1980-06-10 | Research Corporation | Simultaneous immunoassay of multiple antigens and assay for cocaine metabolites |
| US5962336A (en) * | 1997-10-20 | 1999-10-05 | Sun; Ming | Multi-test panel |
| CN101556280A (en) * | 2009-05-20 | 2009-10-14 | 江西中德生物工程有限公司 | Immunochromatography detection card for detecting fluorescent microspheres of ketamine and methyl amphetamine and preparation method thereof |
| CN202854140U (en) * | 2012-06-07 | 2013-04-03 | 北京维德维康生物技术有限公司 | Clenbuterol rapid triple detection card |
| CN103336113A (en) * | 2013-06-24 | 2013-10-02 | 苏州万木春生物技术有限公司 | Preparation method of human immunodeficiency virus (HIV) antibody detection test paper |
| CN105277693A (en) * | 2014-08-18 | 2016-01-27 | 董俊 | Human parainfluenza virus quantum dot immunochromatography typing detection card, preparation method and applications |
| CN104634967A (en) * | 2014-11-30 | 2015-05-20 | 德康润生物科技（天津）有限公司 | Method for further detecting four narcotics in saliva by employing immuno-chromatographic technology |
| CN104655837A (en) * | 2015-02-27 | 2015-05-27 | 南京微测生物科技有限公司 | Fluorescent quantitative test paper strip for simultaneously detecting algal toxins MC-LR/RR/YR and preparation method and application of fluorescent quantitative test paper strip |
| CN107942062A (en) * | 2017-11-29 | 2018-04-20 | 洛阳现代生物技术研究院有限公司 | Test card, preparation and the detection method of synchronous detection 2 toxin of ochracin, vomitoxin and T |
| CN108845151A (en) * | 2018-05-29 | 2018-11-20 | 郑州左安检测科技有限公司 | A kind of test strips and its preparation method and application method detecting crystal methamphetamine |
| CN208607233U (en) * | 2018-06-29 | 2019-03-15 | 江苏达骏生物科技有限公司 | Simultaneous quantitative detects the reagent strip and test card of quinolone, tetracycline and sulfanilamide (SN) |
| CN109490546A (en) * | 2018-12-21 | 2019-03-19 | 广东顺德工业设计研究院(广东顺德创新设计研究院) | Six detection devices of drugs and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| WU, JING等: "Magnetic Lateral Flow Strip for the Detection of Cocaine in Urine by Naked Eyes and Smart Phone Camera", 《SENSORS》 * |
| 曾立波等: "氯胺酮、甲基苯丙胺和吗啡金标单抗试剂盒的研制", 《中国法医学杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110850095A (en) * | 2019-11-22 | 2020-02-28 | 佛山墨赛生物技术有限公司 | Drug trace detection test strip and preparation method and kit thereof |
| CN111474342A (en) * | 2020-04-20 | 2020-07-31 | 石家庄洹众生物科技有限公司 | Colloidal gold detection card for four-item joint detection of drugs and method for detecting drugs |
| CN113391062A (en) * | 2021-05-25 | 2021-09-14 | 黄淮学院 | Morphine immunofluorescence chromatography rapid detection test paper strip, preparation method and detection method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110058012B (en) | 2022-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108181458B (en) | A kind of micro-fluidic chip and its preparation method and application based on fluorescence immunoassay joint-detection | |
| Leute et al. | Spin immunoassay of opiate narcotics in urine and saliva | |
| US20190204309A1 (en) | Point-of-care immunoassay for quantitative small analyte detection | |
| CN110058012A (en) | The fluorescence immunoassay test paper group and its preparation and application of synchronous detection diacetylmorphine, crystal methamphetamine, ketamine and cocaine | |
| US20070254323A1 (en) | Malachite green derivatives for immunoassay reagents to detect malachite green | |
| CN103884682B (en) | Surface plasma body resonant vibration biochip and preparation method and application | |
| CN105652005B (en) | Quantitatively detect time-resolved fluoroimmunoassay chromatograph test strip, the preparation method and applications of Capsaicinoids | |
| CN108508215A (en) | A kind of time-resolved fluoroimmunoassay chromatograph test strip and its preparation method and application of detection tetracycline medication | |
| CN102947703A (en) | Multi epitope assay | |
| CN106084059A (en) | The general specific antibody of anti-Capsaicinoids, test strips and kitchen waste grease immunochromatography method for quick identification | |
| CN109813691A (en) | A kind of time-resolved fluoroimmunoassay chromatograph test strip and its preparation method and application detecting taxanes drug | |
| CN107110847A (en) | For detecting or measuring the ethyl glucuronide aldehydic acid glycosides lateral flow calibration tape of ethyl glucuronide aldehydic acid glycosides, immunoassays, apparatus and method | |
| CN104280437A (en) | Immunosensor and method for detecting various beta-adrenergic receptor stimulant residues | |
| CN104422762A (en) | Drug testing enzyme linked immunosorbent assay kit and detection method thereof | |
| CN108391432A (en) | The Bioconjugation molecule of heterocyclic compound | |
| Cao et al. | Determination of morphine in human urine by the novel competitive fluorescence immunoassay | |
| CN103018440B (en) | A kind of clopidol colloidal gold chromatographic test strip or card | |
| CN1963512A (en) | Chemiluminescence method for qualitative and quantitative detection of hepatitis B virus | |
| CN109444423A (en) | Tetraodotoxin immunofluorescence Rapid detection test strip and preparation method and detection method | |
| CN105624119B (en) | The general monoclonal antibody of anti-capsaicine, Dihydrocapsaicin, the synthetic capsaicin of hybridoma cell strain YQQD8 and its generation | |
| CN101781365A (en) | Artificial antigen of tetraodotoxin and corresponding specific antibody and preparation method and application thereof | |
| CN113109560A (en) | Test strip for quickly testing flonicamid pesticide, preparation method and detection method thereof | |
| CN106460056A (en) | Novel method for detecting detection object in sample, and detection kit using same | |
| CN112250641A (en) | Hydrochlorothiazide hapten, artificial antigen, antibody and preparation method and application thereof | |
| CN102942519B (en) | Nitenpyram hapten, artificial antigen and antibody, their preparation methods and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |