CN111751537A - Test strip, device and method for detecting trace drugs - Google Patents
Test strip, device and method for detecting trace drugs Download PDFInfo
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- CN111751537A CN111751537A CN202010435428.4A CN202010435428A CN111751537A CN 111751537 A CN111751537 A CN 111751537A CN 202010435428 A CN202010435428 A CN 202010435428A CN 111751537 A CN111751537 A CN 111751537A
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- G—PHYSICS
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- 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
- G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
- G01N33/587—Nanoparticles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
- G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
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- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- G—PHYSICS
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- 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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Abstract
The invention relates to a test strip, a device and a method for detecting trace drugs. The test strip comprises a test strip body, wherein the test strip body comprises a substrate, and a sample pad, a combination pad, a detection membrane and a water absorption pad which are arranged on the substrate and are sequentially connected from one end of the substrate to the other end of the substrate; the combination pad is coated with a specific monoclonal antibody of a drug marked by fluorescent microspheres, and the drug comprises morphine and 6-monoacetylmorphine; the detection membrane is provided with a detection line for detecting the specific monoclonal antibody of the drug and a quality control line consisting of quality control protein, and the detection line is coated with a drug antigen for detecting the specific monoclonal antibody of the drug. The method identifies whether the subject takes morphine drugs or not by detecting the combination of morphine and 6-monoacetylmorphine, and has high detection efficiency and reliable result.
Description
Technical Field
The invention relates to the field of drug detection, in particular to a test strip, a device and a method for detecting trace drugs.
Background
At present, most abusers mainly take amphetamine drugs and morphine drugs, wherein: amphetamine-type drugs such as amphetamine, methamphetamine, 3, 4-methylenedioxyamphetamine, 3, 4-methylenedioxymethamphetamine, and the like; examples of morphine-like drugs include 6-monoacetylmorphine and morphine. With the promotion of public security departments to comprehensively developing drug detection work for all employees and public staff in the transportation industry, the entertainment places and the hotel industry, new requirements for drug detection equipment appear. If the detection equipment can simultaneously detect amphetamine drugs and morphine drugs, the detection efficiency of the officers is obviously improved.
The traditional test strip capable of simultaneously detecting amphetamine drugs and morphine drugs sequentially comprises a sample absorption pad, a conjugate release pad, a reaction membrane, a water absorption pad and a bottom plate; the conjugate release pad is sprayed with a methamphetamine antibody, a morphine antibody and a ketamine antibody marked by time-resolved fluorescent microspheres; the reaction membrane is coated with a detection line T1 of methamphetamine antigen, a detection line T2 of morphine antigen, a detection line T3 of ketamine antigen and a quality control line C of goat anti-mouse secondary antibody or Biotin-BSA.
Although the presence of morphine detected in a sample of hair, body fluid, etc. may indicate to some extent that the subject may ingest morphine-like drugs, this is not absolute. Since, when a human body takes an antitussive or analgesic drug, morphine, a component thereof, such as hair, body fluid, etc., can be detected. Therefore, if the conventional test strip is used for detection, it is not favorable to obtain a reliable identification result, and how to improve the reliability of the identification result while realizing efficient detection is an urgent problem to be solved.
Disclosure of Invention
Accordingly, the invention provides a morphine and 6-monoacetylmorphine test strip and a device thereof. When the test strip or the device is used for detection, the reliability of an identification result can be effectively improved while the detection efficiency is ensured.
A test strip for detecting trace drugs comprises a test strip body, wherein the test strip body comprises a substrate, and a sample pad, a combination pad, a detection membrane and a water absorption pad which are arranged on the substrate and are sequentially connected from one end of the substrate to the other end of the substrate;
the combination pad is coated with a specific monoclonal antibody of a drug marked by fluorescent microspheres, and the drug comprises morphine and 6-monoacetylmorphine;
the detection membrane is provided with a detection line for detecting the specific monoclonal antibody of the drug and a quality control line consisting of quality control protein, and the detection line is coated with a drug antigen for detecting the specific monoclonal antibody of the drug.
A preparation method of the test strip comprises the following steps:
separately preparing the conjugate pad and the detection membrane, wherein,
the preparation of the conjugate pad comprises the following steps: preparing a specific monoclonal antibody of the drug marked by the fluorescent microspheres, spraying the monoclonal antibody on a binding pad, and drying;
the preparation of the detection membrane comprises the following steps: respectively taking the drug antigen and the quality control protein, spraying the drug antigen and the quality control protein on a detection membrane, and drying;
assembling the sample pad, conjugate pad, detection membrane, absorbent pad, and the substrate.
The utility model provides a detect device of trace drug, the device includes the casing and as above the test paper strip, the casing includes casing and lower casing, the test paper strip sets up go up the casing with between the casing down, go up the casing and seted up application of sample hole and detection window, application of sample hole corresponds the sample pad of test paper strip, detection window corresponds the detection line and the quality control line of test paper strip.
A method for detecting trace drugs, the method comprising:
and (3) taking a sample to be detected, dripping the sample to the sample pad of the test strip or dripping the sample to the sample adding hole of the device, and carrying out fluorescence detection.
The invention has the following beneficial effects:
according to the test strip and the device provided by the invention, the item 6-monoacetylmorphine is added in morphine detection, and whether a subject takes in opioid drugs or not is identified through the detection of the combination of the two items, so that the detection efficiency is high, and the identification result is reliable. The opioid comprises heroin, codeine, and morphine. Wherein the antitussive liquid contains codeine, and the analgesic contains morphine, codeine, methadone, and dolantin. The heroin is the most common opioid narcotic abused in China, when blood in a body is metabolized, the half life period is 3-9 min, the intermediate metabolite 6-monoacetylmorphine can be converted, the deacetylation of the latter is metabolized into morphine after the half life period is 45min, namely, the 6-monoacetylmorphine is only derived from the heroin, if the intermediate metabolite 6-monoacetylmorphine and the final metabolite morphine are simultaneously used as detection items, an addict can be excluded from evading legal liability by taking cough drops or/and analgesics as reasons, a reliable identification result is obtained, the identification result is strong evidence of opioid narcotic abuse, and the abuser can not avoid shape. The inventors have surprisingly found that when fluorescent microspheres of a suitable inclusion are selected, the relative error between the test results can be reduced.
Drawings
Fig. 1 is a schematic structural diagram of a test strip in embodiment 1 of the present invention;
FIG. 2 is a schematic structural view of an apparatus according to example 1 of the present invention;
figure 3 is a standard curve for morphine in the device of example 3 of the present invention;
FIG. 4 is a standard curve for amphetamines in the apparatus of example 3 of the present invention;
figure 5 is a standard curve for 6-monoacetylmorphine in the device of example 3 of the present invention.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The embodiment of the invention provides a test strip for detecting drugs, which comprises a test strip body, wherein the test strip body comprises a substrate, and a sample pad, a combination pad, a detection membrane and a water absorption pad which are arranged on the substrate and are sequentially connected from one end of the substrate to the other end of the substrate;
the combination pad is coated with a specific monoclonal antibody of a drug marked by fluorescent microspheres, and the drug comprises morphine and 6-monoacetylmorphine;
the detection membrane is provided with a detection line for detecting the specific monoclonal antibody of the drug and a quality control line consisting of quality control protein, and the detection line is coated with a drug antigen for detecting the specific monoclonal antibody of the drug.
Preferably, the fluorescent microspheres are selected from time-resolved fluorescent microspheres or quantum dot fluorescent microspheres, the time-resolved fluorescent microspheres are embedded with europium rare earth elements, and the quantum dot fluorescent microspheres are embedded with CdSe/ZnS.
Preferably, the fluorescent microsphere is wrapped by polystyrene and modified by carboxyl, the particle size is 100nm-400nm, the excitation wavelength is 350nm-400nm, and the emission wavelength is 600nm-630 nm.
It is understood that the drug antigens described in the embodiments of the present invention are all synthetic antigens. Preferably, the morphine antigen is a morphine-conjugated protein complex; and/or the 6-monoacetylmorphine antigen is a 6-monoacetylmorphine-coupled protein complex.
Preferably, the conjugated protein is bovine serum albumin or casein.
Preferably, the quality control protein is selected from rabbit anti-mouse polyclonal antibody IgG or goat anti-mouse IgG. Further preferably, rabbit anti-mouse polyclonal antibody IgG is used.
Preferably, the interval between the detection line and the quality control line and the interval between the adjacent detection lines is 3.5mm-4.5 mm. For example: 3.5mm, 4mm, 4.5 mm. Preferably 3.5mm, 4 mm.
Preferably, the conjugate pad is selected from a fiberglass film or a polyester film; or/and the detection membrane is selected from a cellulose membrane or a cellulose acetate membrane; or/and the substrate is a PVC substrate or other hard non-water-absorbing material substrate.
It is to be understood that the detection line for coating morphine antigen, and the detection line for coating 6-monoacetylmorphine antigen are not limited in order, and preferably, for example, the detection line for coating 6-monoacetylmorphine antigen, the detection line for coating amphetamine drug antigen, and the detection line for coating morphine antigen are sequentially arranged in a direction away from the sample pad.
Preferably, the drugs also include other kinds of drugs. Such as propylamine drugs, ketamine, tetrahydrocannabinol, cocaine, and the like.
Preferably, the other kinds of drugs are phenylpropanoid drugs. The amphetamines in the examples of the present invention refer to a series of synthetic drugs (collectively referred to as amphetamines) that have significant excitatory effects on the central nervous system. It is understood that the following categories are included, but not limited to: amphetamine, methamphetamine, 3, 4-methylenedioxyamphetamine, and 3, 4-methylenedioxymethamphetamine, and the like.
The embodiment of the invention also provides a preparation method of the test strip, which comprises the following steps:
separately preparing the conjugate pad and the detection membrane, wherein,
the preparation of the conjugate pad comprises the following steps: preparing a specific monoclonal antibody of the drug marked by the fluorescent microspheres, spraying the monoclonal antibody on a binding pad, and drying;
the preparation of the detection membrane comprises the following steps: respectively taking the drug antigen and the quality control protein, spraying the drug antigen and the quality control protein on a detection membrane, and drying;
assembling the sample pad, conjugate pad, detection membrane, absorbent pad, and the substrate.
Preferably, the spot concentration of the drug antigen is 0.1mg/mL-1.1mg/mL, and the dosage of the spot is 0.4 μ l/cm-1.0 μ l/cm.
It will be appreciated that the drug antigen may or may not be diluted prior to the spray point, as desired. Preferably, the spot concentration of the drug antigen is 0.1mg/mL-1.1mg/mL, and the dosage of the spot is 0.4 μ l/cm-1.0 μ l/cm.
Preferably, the spot concentration of the amphetamine antigen is 0.2mg/mL-0.6mg/mL, the spot concentration of the morphine antigen is 0.1mg/mL-0.5mg/mL, the spot concentration of the 6-monoacetylmorphine antigen is 0.2mg/mL-0.6mg/mL, the concentration of the dilution of the quality control protein is 0.1mg/mL-1.1mg/mL, and the dosage of the spot is 0.4 mu l/cm-1.0 mu l/cm.
Preferably, the step of preparing the fluorescent microsphere labeled drug-specific monoclonal antibody comprises:
taking the fluorescent microspheres, adding a coupling agent, incubating, centrifuging, removing a supernatant, dispersing in an activation buffer solution, and preparing a mixed solution a;
taking the drug specific monoclonal antibody, dispersing the drug specific monoclonal antibody in a coupling buffer solution to prepare a mixed solution b;
adding the dispersion liquid a into the dispersion liquid b, mixing and shaking to prepare a mixed liquid c;
centrifuging the mixed solution c, removing a supernatant, adding a confining liquid, and oscillating to prepare a mixed solution d;
and (4) centrifuging the mixed solution d, and removing the supernatant.
Preferably, in the step of preparing the mixed solution a: the mass concentration of the fluorescent microspheres is 10-40%.
Preferably, in the step of preparing the mixed solution a: the coupling agent is selected from EDC or/and the incubation is performed at 24-28 ℃ for 0.3-0.6 h; or/and the centrifugation is frozen centrifugation at 12000rpm-15000rpm for 8min-12 min.
Preferably, in the step of preparing the mixed solution c, the shaking is carried out at 25-37 ℃ for 0.5-2 h.
Preferably, in the step of preparing the mixed solution d: the centrifugation is frozen and centrifuged for 5min to 10min at 8000rpm to 10000 rpm; or/and the shaking is carried out for 0.5h-2h at the temperature of 25 ℃ -37 ℃.
Preferably, the centrifugation of the mixed solution d is refrigerated centrifugation at 8000-10000 rpm for 5-10 min.
The embodiment of the invention also provides a device for detecting drugs, which comprises a shell and the test strip, wherein the shell comprises an upper shell and a lower shell, the test strip is arranged between the upper shell and the lower shell, the upper shell is provided with a sample adding hole and a detection window, the sample adding hole corresponds to a sample pad of the test strip, and the detection window corresponds to a detection line and a quality control line of the test strip.
Preferably, one end of the upper shell, which is far away from the sample adding hole, is provided with a handheld part. Such a design is more advantageous to operate.
Preferably, the device is also provided with an information label. It is understood that the information label may be a bar code, two-dimensional code, etc. with information including, but not limited to, standard curves and production lot numbers. In fact, information such as the validity period can be embedded in the information tag according to needs. It is to be understood that the information label may be disposed on the upper case surface or the lower case surface without limitation.
The embodiment of the invention also provides a drug detection method, which comprises the following steps:
and (3) taking a sample to be detected, dripping the sample to the sample pad of the test strip or dripping the sample to the sample adding hole of the device, and carrying out fluorescence detection.
Preferably, the sample to be tested is selected from at least one of blood, saliva, urine, sweat and hair. It will be understood that the test sample described herein may be directly loaded for measurement, or may be subjected to some type of pretreatment, such as dilution, lysis, etc., followed by loading for measurement.
According to the detection requirement, the test strip and the device provided by the embodiment of the invention can be matched with a diluent for use, the diluent is preferably a diluent which can stably store drugs for at least 1 to 2 years at normal temperature, and has a rapid cracking effect on hair, and the sample can be loaded after cracking and standing for 3 to 5 min.
The following examples of the invention: time-resolved fluorescent microspheres, purchased from Shanghai tracing Biotechnology Ltd; the quantum dot fluorescent microspheres are purchased from Shanghai Kun-Dai Biotechnology Limited; an activation buffer solution with 10 mM-100 mM MES pH 5-7; coupling buffer solution, 10 mM-50 mM PBS; a storage solution, 10mM of Tirs-HCl PH 7-9, and 4 ten thousand of 2% glucan; spraying a membrane buffer solution, 10mM of Tirs-HCl, pH 7.5-8.5, 2% of D-trehalose and 0.5% -1% of S9; sample diluent 10 mM-20 mMPBS, 0.5% -2% OVA and 0.1% -0.9% NaCl.
As shown in fig. 1, the test strip of the present embodiment includes a test strip body, and the test strip body includes a substrate 5, and a sample pad 1, a conjugate pad 2, a detection membrane 3, and a water absorption pad 4, which are disposed on the substrate 5 and connected in sequence from one end of the substrate 5 to the other end; wherein: the combination pad 2 is a glass fiber membrane, the detection membrane 3 is a nitrocellulose membrane, and the substrate 5 is a PVC rubber plate;
the combination pad 2 is coated with a drug-specific monoclonal antibody marked by fluorescent microspheres, and the drug-specific monoclonal antibody comprises a phenylpropylamine-specific monoclonal antibody, a morphine-specific monoclonal antibody and a 6-monoacetylmorphine-specific antibody; wherein: the fluorescent microsphere adopts a time-resolved fluorescent microsphere, is embedded with europium rare earth elements, is wrapped by polystyrene and modified by carboxyl, and has the particle size of 100nm, the excitation wavelength of 360nm and the emission wavelength of 615 nm.
The detection membrane 3 is provided with a detection line 6 and a quality control line 7(C line) formed by quality control protein, wherein the detection line 6 comprises a detection line (T1 line) for coating 6-monoacetylmorphine antigen, a detection line T2(T2 line) for coating amphetamine antigen and a detection line (T3 line) for coating morphine antigen; wherein: the 6-monoacetylmorphine antigen is a 6-monoacetylmorphine-BSA compound, the amphetamine antigen is an amphetamine-BSA compound, the morphine antigen is a morphine-BSA compound, and the quality control protein is a rabbit anti-mouse polyclonal antibody; the detection line and the quality control line are arranged in parallel on the detection membrane 3 from the sample pad to the absorbent pad according to the sequence of a T1 line, a T2 line, a T3 line and a C line, and the interval between two adjacent lines is 4 mm.
As shown in fig. 2, the device for detecting drugs in this embodiment includes a housing and the above test strip, where the housing includes an upper housing and a lower housing, the test strip is disposed between the upper housing and the lower housing, the upper housing is provided with a sample hole 8 and a detection window 9, the sample hole 8 corresponds to a sample pad 1 of the test strip, and the detection window 9 corresponds to a detection line and a quality control line on a detection membrane 3 of the test strip; a handheld part 10 is arranged at one end of the upper shell, which is far away from the sample adding hole; the device is also provided with a bar code 11, and the bar code is provided with a standard curve and production batch number information.
Example 2 and example 1 preparation of test paper strips
The embodiment provides a preparation method of a test strip for detecting amphetamines, morphine and 6-monoacetylmorphine, which comprises the following steps:
A. preparing the drug specific monoclonal antibody marked by the fluorescent microspheres, diluting, spraying the diluted drug specific monoclonal antibody on a binding pad, drying, and preparing the binding pad coated with the drug specific monoclonal antibody marked by the fluorescent microspheres. The method comprises the following specific steps:
a1, preparing a fluorescent microsphere labeled drug specific monoclonal antibody:
(a) preparing the fluorescent microspheres into 10% of working concentration;
(b) adding 2 μ l of coupling agent EDC solution (containing EDC with concentration of 20mg/mL), mixing, and incubating at room temperature for 0.5 h; carrying out refrigerated centrifugation at 12000rpm for 10min, removing supernatant, dispersing the fluorescent microspheres in 100 mu l of activation buffer solution, and carrying out uniform ultrasonic dispersion;
(c) then adding 100 mu g of drug specific antibody, dispersing in 100 mu l of coupling buffer solution, adding the fluorescent microspheres into the drug specific antibody, uniformly mixing, keeping the temperature at 25 ℃, shaking and uniformly mixing for 2 h;
(d) performing refrigerated centrifugation at 10000rpm for 10min to remove free drug specific antibodies;
(e) adding 200 mul of BSA blocking solution with the mass concentration of 1%, performing ultrasonic dispersion, keeping the temperature at 25 ℃, and shaking and mixing uniformly for 2 h;
(f) the mixture was frozen and centrifuged at 10000rpm for 10min, and 200. mu.l of the stock solution was added.
A2 preparation of bonding pad
(1) Taking the amphetamine monoclonal antibody, the morphine monoclonal antibody and the 6-monoacetylmorphine monoclonal antibody which are marked by the fluorescent microspheres prepared in the step A1;
(2) mixing the amphetamine monoclonal antibody, the morphine monoclonal antibody and the 6-monoacetylmorphine monoclonal antibody in the step (1) according to the ratio of 2:3: 2;
(3) diluting the mixture obtained in the step (2) by adopting a membrane spraying buffer solution, spraying the mixture on a glass fiber pad treated by 0.25-2% PVP5.8 ten thousand solution, and drying to prepare a bonding pad (also called an immunofluorescence pad); wherein the dilution factor of the mixture obtained in the step (2) by the membrane spraying buffer solution is 5 times.
B. Respectively taking the amphetamine antigen, the morphine antigen, the 6-monoacetylmorphine antigen and the quality control protein, diluting, spraying the diluted solution on a detection membrane, drying, and manufacturing the detection membrane provided with a detection line and a quality control line. The method specifically comprises the following steps:
spraying 6-monoacetylmorphine-BSA (with the spraying point concentration of 0.2mg/mL), amphetamine-BSA (with the spraying point concentration of 0.2mg/mL), morphine-BSA (with the spraying point concentration of 0.1mg/mL) and rabbit anti-mouse polyclonal antibody (with the spraying point concentration of 0.1mg/mL) on one end, close to the binding pad, of the detection membrane in sequence to form a T1 line, a T2 line, a T3 line and a C line correspondingly; the spot size was 0.4. mu.l/cm.
C. Assembling the sample pad, conjugate pad, detection membrane, absorbent pad, and the substrate. The method comprises the following specific steps:
and (3) sequentially sticking the sample pad, the bonding pad (also called an immunofluorescence pad) prepared in the step (A), the detection membrane and the water absorption pad prepared in the step (B) onto a PVC rubber plate to prepare a large plate, and cutting to prepare the single-person detection reagent strip.
And finally, filling the detection reagent strip into a plastic card shell (between the upper shell and the lower shell) to prepare the device for detecting the drugs.
The device of the embodiment can be placed in an aluminum foil bag together with a matched sample diluent, a quantitative dropper and a drying agent, sealed by a sealing machine and placed and stored at normal temperature. The detection sensitivity, precision, thermal stability, low-temperature stability of testing environment, stability of testing time and specificity before delivery.
Example 3
Establishing standard curves of amphetamines, morphine and 6-monoacetylmorphine, and performing precision detection, wherein the implementation method comprises the following steps:
(1) establishing a standard curve
Morphine enterprise reference, amphetamine enterprise reference and 6-monoacetylmorphine enterprise reference are respectively diluted by sample diluent matched with the enterprise to obtain nine concentrations of 0ng/mg, 0.05ng/mg, 0.1ng/mg, 0.2ng/mg, 0.5ng/mg, 1.0ng/mg, 2.0ng/mg, 5.0ng/mg and 10 ng/mg.
The assay was performed by loading 80. mu.L of sample using the prepared apparatus of example 2, and the average was taken by repeating 4 times for each concentration. The fluorescence intensity of the detection window of the detection device of the matched fluorescence analyzer is used, and the relationship of the concentration corresponding to the fluorescence intensity is obtained by adopting the moving average fitting of the ExCel table. As shown in fig. 3,4 and 5.
(2) Detection of precision
The morphine business reference, amphetamine business reference and 6-monoacetylmorphine business reference prepared in example 2 were diluted as shown in (1) to concentrations of 0ng/mg, 0.1ng/mg and 1.0ng/mg, respectively, and each concentration was tested in duplicate for 10 times, and coefficient of variation CV was calculated. The results are shown in tables 1, 2 and 3 below.
TABLE 1 measurement of the precision of different morphine concentrations
TABLE 2 measurement of the precision of the amphetamine-type compounds at different concentrations
| Test concentration | 0ng/mg | 0.1ng/mg | 1.0ng/ |
| Test | |||
| 1 | <0.05 | 0.10 | 1.11 |
| |
<0.05 | 0.10 | 1.05 |
| Test 3 | <0.05 | 0.09 | 0.93 |
| |
<0.05 | 0.09 | 1.07 |
| Test 5 | <0.05 | 0.10 | 0.95 |
| |
<0.05 | 0.11 | 0.81 |
| Test 7 | <0.05 | 0.09 | 1.01 |
| |
<0.05 | 0.08 | 1.12 |
| |
<0.05 | 0.10 | 1.01 |
| |
<0.05 | 0.10 | 0.99 |
| Coefficient of |
0 | 8.78％ | 9.23％ |
TABLE 3 precision measurement of different concentrations of 6-monoacetylmorphine
| Test concentration | 0ng/mg | 0.1ng/mg | 1.0ng/ |
| Test | |||
| 1 | <0.05 | 0.09 | 1.00 |
| |
<0.05 | 0.08 | 1.02 |
| Test 3 | <0.05 | 0.10 | 0.88 |
| |
<0.05 | 0.09 | 1.03 |
| Test 5 | <0.05 | 0.10 | 1.04 |
| |
<0.05 | 0.09 | 0.93 |
| Test 7 | <0.05 | 0.11 | 0.99 |
| |
<0.05 | 0.10 | 1.00 |
| |
<0.05 | 0.10 | 1.15 |
| |
<0.05 | 0.10 | 1.04 |
| Coefficient of |
0 | 8.78％ | 7.07％ |
As can be seen from tables 1, 2 and 3, when 0ng/mg was measured, the fluorescence analyzer automatically reported <0.05ng/mg and judged negative. When the test concentration is 0.1ng/mg and 1.0ng/mg, the variation coefficients of the three detection items are all less than 10%, and the product requirements are met.
Example 4
The device prepared in example 2 was used to compare the effects of test temperatures in different environments on the test results, and the experimental method was as follows:
cutting suspected positive and negative hair sample with scissors, adding into 5 mg-800 μ L of the sample diluent, mixing, standing for cracking for 5min, and adding 80 μ L of sample at 5 deg.C, 10 deg.C, 15 deg.C, 20 deg.C, 25 deg.C, 30 deg.C, 35 deg.C, 40 deg.C for detection. The measurement results and the standard deviation RSD with respect to 25 ℃ were calculated, and the results are shown in tables 4, 5, and 6 below.
TABLE 4 comparison of temperature of morphine at different environmental tests
| Test temperature | Negative of | RSD | Suspected positivity | RSD |
| 5℃ | <0.05 | 0.00％ | 4.11 | -11.99％ |
| 10℃ | <0.05 | 0.00％ | 3.99 | -14.56％ |
| 15℃ | <0.05 | 0.00％ | 5.01 | 7.28％ |
| 20℃ | <0.05 | 0.00％ | 4.42 | -5.35％ |
| 25℃ | <0.05 | 0.00％ | 4.67 | 0.00％ |
| 30℃ | <0.05 | 0.00％ | 4.52 | -3.21％ |
| 35℃ | <0.05 | 0.00％ | 4.88 | 4.50％ |
| 40℃ | <0.05 | 0.00％ | 4.95 | 6.00％ |
TABLE 5 comparison of temperatures tested in different environments for amphetamines
| Test temperature | Negative of | RSD | Suspected positivity | RSD |
| 5℃ | <0.05 | 0.00％ | 2.21 | -14.67％ |
| 10℃ | <0.05 | 0.00％ | 2.96 | 14.29％ |
| 15℃ | <0.05 | 0.00％ | 2.89 | 11.58％ |
| 20℃ | <0.05 | 0.00％ | 2.33 | -10.04％ |
| 25℃ | <0.05 | 0.00％ | 2.59 | 0.00％ |
| 30℃ | <0.05 | 0.00％ | 2.61 | 0.77％ |
| 35℃ | <0.05 | 0.00％ | 2.74 | 5.79％ |
| 40℃ | <0.05 | 0.00％ | 2.86 | 10.42％ |
TABLE 6 comparison of temperature of 6-monoacetylmorphine at different environmental tests
As can be seen from tables 4, 5, and 6, when a negative sample is tested, the fluorescence analyzer automatically reports <0.05ng/mg, and the negative coincidence rate of all three items is 100%, and when a positive sample is tested, the relative deviation is less than 15%, indicating that the device can be used for detection in the ambient temperature range of 5 ℃ to 40 ℃.
Example 5
The device prepared in example 2 was used to compare the effect of different test times on the test results, and the experimental method was as follows:
cutting suspected positive and negative hair samples with scissors, adding into 5 mg-800 μ L of the supporting sample diluent, mixing, standing and cracking for 5min, and adding 80 μ L of sample for detection at 1min, 3min, 5min, 10min, 15min, 20min, 25min, 30min, 35min, and 40 min. The measurement results and the standard deviation RSD with respect to 5min were calculated, and the results are shown in tables 7, 8, and 9 below.
TABLE 7 comparison of morphine for different test times
| Time of measurement | Negative of | RSD | Suspected positivity | RSD |
| 1min | <0.05 | 0.00％ | 3.18 | 25.35％ |
| 3min | <0.05 | 0.00％ | 4.33 | -1.64％ |
| 5min | <0.05 | 0.00％ | 4.26 | 0.00％ |
| 10min | <0.05 | 0.00％ | 4.51 | -5.87％ |
| 15min | <0.05 | 0.00％ | 4.69 | -10.09％ |
| 20min | <0.05 | 0.00％ | 4.55 | -6.81％ |
| 25min | <0.05 | 0.00％ | 4.88 | -14.55％ |
| 30min | <0.05 | 0.00％ | 4.73 | -11.03％ |
| 35min | 0.08 | 60.0％ | 5.11 | -19.95％ |
| 40min | <0.05 | 0.00％ | 5.39 | -26.53％ |
TABLE 8 comparison of different test times for amphetamines
TABLE 9 comparison of different test times for 6-monoacetylmorphine
| Time of measurement | Negative of | RSD | Suspected positivity | RSD |
| 1min | 0.10 | 100.0％ | 5.23 | -39.47％ |
| 3min | <0.05 | 0.00％ | 3.96 | -5.60％ |
| 5min | <0.05 | 0.00％ | 3.75 | 0.00％ |
| 10min | <0.05 | 0.00％ | 4.13 | -10.13％ |
| 15min | <0.05 | 0.00％ | 4.21 | -12.27％ |
| 20min | <0.05 | 0.00％ | 3.33 | 11.20％ |
| 25min | <0.05 | 0.00％ | 4.02 | -7.20％ |
| 30min | <0.05 | 0.00％ | 3.89 | -3.73％ |
| 35min | <0.05 | 0.00％ | 4.35 | -16.00％ |
| 40min | 0.08 | 60.0％ | 4.57 | -21.87％ |
As can be seen from tables 7, 8 and 9, the test times for three items of morphine, amphetamines and 6-monoacetylmorphine can be performed within a range of 3min to 30min after sample lysis with a relative deviation of less than 15%.
As shown in fig. 1, the test strip of the present embodiment includes a test strip body, and the test strip body includes a substrate 5, and a sample pad 1, a conjugate pad 2, a detection membrane 3, and a water absorption pad 4, which are disposed on the substrate 5 and connected in sequence from one end of the substrate 5 to the other end; wherein: the combination pad 2 is a polyester film, the detection film 3 is a cellulose acetate film, and the substrate 5 is a PVC rubber plate;
the combination pad 2 is coated with a drug-specific monoclonal antibody marked by fluorescent microspheres, and the drug-specific monoclonal antibody comprises a phenylpropylamine-specific monoclonal antibody, a morphine-specific monoclonal antibody and a 6-monoacetylmorphine-specific antibody; wherein: the fluorescent microsphere adopts quantum dot fluorescent microsphere, is embedded with CdSe/ZnS, is wrapped by polystyrene and modified by carboxyl, and has particle size of 200nm, excitation wavelength of 390nm and emission wavelength of 600 nm.
The detection membrane 3 is provided with a detection line 6 and a quality control line 7(C line) formed by quality control protein, wherein the detection line 6 comprises a detection line (T1 line) for coating 6-monoacetylmorphine antigen, a detection line T2(T2 line) for coating amphetamine antigen and a detection line (T3 line) for coating morphine antigen; wherein: the 6-monoacetylmorphine antigen is a 6-monoacetylmorphine-BSA compound, the amphetamine antigen is an amphetamine-BSA compound, the morphine antigen is a morphine-BSA compound, and the quality control protein is a rabbit anti-mouse polyclonal antibody; the detection line and the quality control line are arranged in parallel on the detection membrane 3 from the sample pad to the absorbent pad according to the sequence of a T1 line, a T2 line, a T3 line and a C line, and the interval between two adjacent lines is 3.5 mm.
As shown in fig. 2, the device of this embodiment includes a housing and the above test strip, where the housing includes an upper housing and a lower housing, the test strip is disposed between the upper housing and the lower housing, the upper housing is provided with a sample hole 8 and a detection window 9, the sample hole 8 corresponds to a sample pad 1 of the test strip, and the detection window 9 corresponds to a detection line and a quality control line on a detection membrane 3 of the test strip; a handheld part 10 is arranged at one end of the upper shell, which is far away from the sample adding hole; the device is also provided with a bar code 11, and the bar code is provided with a standard curve and production batch number information.
Example 7 and example 6 preparation of test strips
The embodiment provides a preparation method of a test strip for detecting amphetamines, morphine and 6-monoacetylmorphine, which comprises the following steps:
A. preparing the drug specific monoclonal antibody marked by the fluorescent microspheres, diluting, spraying the diluted drug specific monoclonal antibody on a binding pad, drying, and preparing the binding pad coated with the drug specific monoclonal antibody marked by the fluorescent microspheres. The method comprises the following specific steps:
a1, preparing a fluorescent microsphere labeled drug specific monoclonal antibody:
(a) preparing the fluorescent microspheres into a working concentration of 40%;
(b) adding 10 μ l of coupling agent EDC solution (containing EDC with concentration of 15mg/mL), mixing, and incubating at room temperature for 0.5 h; freezing and centrifuging at 15000rpm for 12min, removing supernatant, dispersing fluorescent microspheres in 500 μ l of activation buffer, and ultrasonically dispersing uniformly;
(c) then adding 100 mu g of drug specific antibody, dispersing in 1000 mu l of coupling buffer solution, adding the fluorescent microspheres into the drug specific antibody, uniformly mixing, keeping the temperature at 37 ℃, shaking and uniformly mixing for 0.5 h;
(d) carrying out refrigerated centrifugation at 8000rpm for 5min to remove free drug specific antibody;
(e) adding 1000 mul of BSA blocking solution with the mass concentration of 1%, ultrasonically dispersing, keeping the temperature at 37 ℃, shaking and uniformly mixing for 0.5 h;
(f) the mixture was frozen and centrifuged at 8000rpm for 5min, and then the supernatant was removed and 1000. mu.l of the stock solution was added.
A2 preparation of bonding pad
(1) Taking the amphetamine monoclonal antibody, the morphine monoclonal antibody and the 6-monoacetylmorphine monoclonal antibody which are marked by the fluorescent microspheres prepared in the step A1;
(2) mixing the amphetamine monoclonal antibody, the morphine monoclonal antibody and the 6-monoacetylmorphine monoclonal antibody in the step (1) according to the ratio of 2:3: 2;
(3) diluting the mixture obtained in the step (2) by adopting a membrane spraying buffer solution of 5% glucose and 1% BSA, spraying the mixture on a glass fiber pad pretreated by 4% PVP5.8, and drying to prepare a bonding pad (also called an immunofluorescence pad); wherein the dilution factor of the mixture obtained in the step (2) by the membrane spraying buffer solution is 20 times.
B. Respectively taking the amphetamine antigen, the morphine antigen, the 6-monoacetylmorphine antigen and the quality control protein, diluting, spraying the diluted solution on a detection membrane, drying, and manufacturing the detection membrane provided with a detection line and a quality control line. The method specifically comprises the following steps:
spraying 6-monoacetylmorphine-BSA (with the spraying point concentration of 0.6mg/mL), amphetamine-BSA (with the spraying point concentration of 0.6mg/mL), morphine-BSA (with the spraying point concentration of 0.5mg/mL) and rabbit anti-mouse polyclonal antibody (with the spraying point concentration of 1.1mg/mL) on one end, close to the binding pad, of the detection membrane in sequence to form a T1 line, a T2 line, a T3 line and a C line correspondingly; the amount of the sprayed dots was 1.0. mu.l/cm.
C. Assembling the sample pad, conjugate pad, detection membrane, absorbent pad, and the substrate. The method comprises the following specific steps:
and (3) sequentially sticking the sample pad, the bonding pad (also called an immunofluorescence pad) prepared in the step (A), the detection membrane and the water absorption pad prepared in the step (B) onto a PVC rubber plate to prepare a large plate, and cutting to prepare the single-person detection reagent strip.
And finally, filling the detection reagent strip into a plastic card shell (between the upper shell and the lower shell) to prepare the device for detecting the drugs.
The device of the embodiment can be placed in an aluminum foil bag together with a matched sample diluent, a quantitative dropper and a drying agent, sealed by a sealing machine and placed and stored at normal temperature. The detection sensitivity, precision, thermal stability, low-temperature stability of testing environment, stability of testing time and specificity before delivery.
Example 8
This example is a variation of example 1, and the main variation of the apparatus of this example with respect to the apparatus of example 1 is that the time-resolved fluorescent microspheres are embedded with lanthanide rare earth elements.
And (3) shearing suspected positive and negative hair samples by using scissors at room temperature, adding the hair samples into a matched sample diluent of which the volume is 5mg to 800 mu L, uniformly mixing, standing and cracking for 5min, and adding 80 mu L of the sample into a device for detection after 10 min.
As a result: negative samples, 5min test and 10min test results are negative; when the positive sample is detected within the range of 3min-30min, the relative error is more than 15%.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (15)
1. The test strip for detecting the trace drugs is characterized by comprising a test strip body, wherein the test strip body comprises a substrate, and a sample pad, a combination pad, a detection membrane and a water absorption pad which are arranged on the substrate and are sequentially connected from one end of the substrate to the other end of the substrate;
the combination pad is coated with a specific monoclonal antibody of a drug marked by fluorescent microspheres, and the drug comprises morphine and 6-monoacetylmorphine;
and the detection membrane is provided with a detection line and a quality control line consisting of quality control protein, and the detection line is coated with corresponding drug antigens.
2. The test strip of claim 1, wherein the fluorescent microspheres are selected from time-resolved fluorescent microspheres embedded with europium rare earth elements or quantum dot fluorescent microspheres embedded with CdSe/ZnS.
3. The test strip of claim 2, wherein the fluorescent microspheres are coated with polystyrene and modified by carboxyl groups, and have a particle size of 100nm-400nm, an excitation wavelength of 350nm-400nm, and an emission wavelength of 600nm-630 nm.
4. The test strip of any one of claims 1 to 3, wherein the drug antigen for detecting a monoclonal antibody specific for morphine is a morphine-conjugated protein complex; and/or the drug antigen for detecting the 6-monoacetylmorphine-specific monoclonal antibody is a 6-monoacetylmorphine coupling protein complex.
5. The test strip of claim 4, wherein the conjugated protein is bovine serum albumin or casein.
6. The test strip of any one of claims 1 to 3 and 5, wherein the quality control protein is selected from a rabbit anti-mouse polyclonal antibody or a goat anti-mouse;
or/and the interval between the detection line and the quality control line and the interval between the adjacent detection lines are 3.5mm-4.5 mm;
or/and, the conjugate pad is selected from a fiberglass film or a polyester film;
or/and the detection membrane is selected from a nitrocellulose membrane or a cellulose acetate membrane;
or/and the substrate is a PVC substrate or other hard non-water-absorbing material substrate;
or/and a detection line for detecting the monoclonal antibody of 6-monoacetylmorphine, a detection line for detecting the monoclonal antibody of morphine and the quality control line are sequentially arranged on the detection membrane from one end of the sample pad to one end of the absorbent pad;
and/or the drugs also comprise other drugs.
7. The test strip of claim 6, wherein the other types of drugs are amphetamine-type drugs.
8. A method for preparing the test strip of any one of claims 1 to 7, wherein the method comprises the following steps:
separately preparing the conjugate pad and the detection membrane, wherein,
the preparation of the conjugate pad comprises the following steps: preparing a specific monoclonal antibody of the drug marked by the fluorescent microspheres, spraying the monoclonal antibody on a binding pad, and drying;
the preparation of the detection membrane comprises the following steps: respectively taking the drug antigen and the quality control protein, spraying the drug antigen and the quality control protein on a detection membrane, and drying;
assembling the sample pad, conjugate pad, detection membrane, absorbent pad, and the substrate.
9. The method of claim 8, wherein the spot concentration of the drug antigen is 0.1mg/mL to 1.1mg/mL, and the spot dosage is 0.4 μ l/cm to 1.0 μ l/cm.
10. The method for preparing the test strip of claim 8 or 9, wherein the step of preparing the monoclonal antibody specific to the drug labeled by the fluorescent microsphere comprises:
taking the fluorescent microspheres, adding a coupling agent, incubating, centrifuging, removing a supernatant, dispersing in an activation buffer solution, and preparing a mixed solution a;
taking the drug specific monoclonal antibody, dispersing the drug specific monoclonal antibody in a coupling buffer solution to prepare a mixed solution b;
adding the dispersion liquid a into the dispersion liquid b, mixing and shaking to prepare a mixed liquid c;
centrifuging the mixed solution c, removing a supernatant, adding a confining liquid, and oscillating to prepare a mixed solution d;
and (4) centrifuging the mixed solution d, and removing the supernatant.
11. The method for preparing the test strip of claim 10, wherein the step of preparing the mixed solution a comprises: the mass concentration of the fluorescent microspheres is 10-40%; or/and, the coupling agent is selected from EDC; or/and the incubation is carried out for 0.3h to 0.6h at the temperature of 24 ℃ to 28 ℃; or/and the centrifugation is frozen centrifugation at 12000rpm-15000rpm for 8min-12 min;
or/and in the step of preparing the mixed solution c, the shaking is carried out for 0.5 to 2 hours at the temperature of between 25 and 37 ℃;
or/and in the preparation step of the mixed solution d, the centrifugation is refrigerated and centrifuged for 5min to 10min at 8000rpm to 10000 rpm; or/and the shaking is carried out for 0.5h-2h at the temperature of 25 ℃ -37 ℃;
or/and the step of centrifuging the mixed solution d is to freeze and centrifuge at 8000rpm-10000rpm for 5min-10 min.
12. A device for detecting trace drugs, comprising a housing and the test strip of any one of claims 1 to 7, wherein the housing comprises an upper housing and a lower housing, the test strip is disposed between the upper housing and the lower housing, the upper housing is provided with a sample hole and a detection window, the sample hole corresponds to a sample pad of the test strip, and the detection window corresponds to a detection line and a quality control line of the test strip.
13. The device of claim 12, wherein a hand-held portion is provided at an end of the upper housing away from the sample application hole;
or/and the device is also provided with an information label.
14. A method for detecting trace drugs, comprising:
dropping a sample to be detected on the sample pad of the test strip of any one of claims 1 to 7 or in the sample hole of the device of claim 12 or 13, and carrying out fluorescence detection.
15. The method according to claim 14, wherein the sample to be tested is at least one selected from blood, saliva, urine, sweat, and hair.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112698027A (en) * | 2020-12-14 | 2021-04-23 | 广东唯实生物技术有限公司 | Hapten immunochromatography detection reagent |
| CN113049827A (en) * | 2021-03-12 | 2021-06-29 | 郑州大学 | Time-resolved fluorescence immunochromatography reagent strip, preparation method and hemp concentration quantitative detection method |
| 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 |
| CN113640521A (en) * | 2021-06-25 | 2021-11-12 | 广东省大湾区华南理工大学聚集诱导发光高等研究院 | Test strip and kit for quantitatively determining drugs in hair, preparation method and application |
| CN114137232A (en) * | 2021-11-19 | 2022-03-04 | 佛山墨赛生物技术有限公司 | Latex microsphere immunoassay test strip, kit and method |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103207242A (en) * | 2012-01-11 | 2013-07-17 | 司法部司法鉴定科学技术研究所 | Method for rapidly identifying monoacetylmorphine and morphine in hair |
| CN104422762A (en) * | 2013-09-11 | 2015-03-18 | 杭州优玛达生物科技有限公司 | Drug testing enzyme linked immunosorbent assay kit and detection method thereof |
| CN105916860A (en) * | 2013-03-14 | 2016-08-31 | 美艾利尔圣地亚哥公司 | 6-acetylmorphine analogs, and methods for their synthesis and use |
| CN108844922A (en) * | 2018-05-17 | 2018-11-20 | 浙江诺迦生物科技有限公司 | The rapid detection method of drugs in a kind of hair |
| CN110208072A (en) * | 2019-05-10 | 2019-09-06 | 江苏苏博生物医学科技南京有限公司 | It is a kind of for the human hair rapid cleavage liquid of trace illicit drugs inspection and its application |
| 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 |
| CN110927381A (en) * | 2019-12-04 | 2020-03-27 | 郑州左安检测科技有限公司 | Test strip for detecting 6-monoacetylmorphine and preparation method and application method thereof |
-
2020
- 2020-05-21 CN CN202010435428.4A patent/CN111751537A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103207242A (en) * | 2012-01-11 | 2013-07-17 | 司法部司法鉴定科学技术研究所 | Method for rapidly identifying monoacetylmorphine and morphine in hair |
| CN105916860A (en) * | 2013-03-14 | 2016-08-31 | 美艾利尔圣地亚哥公司 | 6-acetylmorphine analogs, and methods for their synthesis and use |
| CN104422762A (en) * | 2013-09-11 | 2015-03-18 | 杭州优玛达生物科技有限公司 | Drug testing enzyme linked immunosorbent assay kit and detection method thereof |
| CN108844922A (en) * | 2018-05-17 | 2018-11-20 | 浙江诺迦生物科技有限公司 | The rapid detection method of drugs in a kind of hair |
| CN110208072A (en) * | 2019-05-10 | 2019-09-06 | 江苏苏博生物医学科技南京有限公司 | It is a kind of for the human hair rapid cleavage liquid of trace illicit drugs inspection and its application |
| 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 |
| CN110927381A (en) * | 2019-12-04 | 2020-03-27 | 郑州左安检测科技有限公司 | Test strip for detecting 6-monoacetylmorphine and preparation method and application method thereof |
Non-Patent Citations (5)
| Title |
|---|
| PRIYA MISHRA 等: "An immunochromatographic dipstick as an alternate for monitoring of heroin metabolites in urine samples", 《RSC ADVANCES》 * |
| SONU GANDHI 等: "A gold nanoparticle-single-chain fragment variable antibody as an immunoprobe for rapid detection of morphine by dipstick", 《RSC ADVANCES》 * |
| 刘佳 等: "吗啡免疫荧光层析快速定量方法的建立", 《中国药学杂志》 * |
| 康可人 等: "应用于侧向免疫层析方法学的高特异性抗吗啡单克隆抗体的制备", 《中国药物依赖性杂志》 * |
| 陈雅倩 等: "海洛因及其代谢物吗啡的分析研究进展", 《化学传感器》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112698027A (en) * | 2020-12-14 | 2021-04-23 | 广东唯实生物技术有限公司 | Hapten immunochromatography detection reagent |
| CN113049827A (en) * | 2021-03-12 | 2021-06-29 | 郑州大学 | Time-resolved fluorescence immunochromatography reagent strip, preparation method and hemp concentration quantitative detection method |
| 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 |
| CN113391062B (en) * | 2021-05-25 | 2024-06-07 | 黄淮学院 | Morphine immunofluorescence chromatography rapid detection test strip, preparation method and detection method thereof |
| CN113640521A (en) * | 2021-06-25 | 2021-11-12 | 广东省大湾区华南理工大学聚集诱导发光高等研究院 | Test strip and kit for quantitatively determining drugs in hair, preparation method and application |
| CN114137232A (en) * | 2021-11-19 | 2022-03-04 | 佛山墨赛生物技术有限公司 | Latex microsphere immunoassay test strip, kit and method |
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