CN118150823A - Etomidate immunofluorescence chromatography rapid detection test paper and detection method - Google Patents
Etomidate immunofluorescence chromatography rapid detection test paper and detection method Download PDFInfo
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Classifications
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- G—PHYSICS
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- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- 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
- G01N33/577—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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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
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- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses etomidate immunofluorescence chromatography rapid detection test paper and a detection method, and relates to the technical fields of qualitative identification and drug administration detection. The etomidate immunofluorescence chromatography rapid detection test paper comprises a bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad; the nitrocellulose membrane is attached to the bottom plate, the water absorbing pad and the bonding pad are respectively tightly pressed against two ends of the nitrocellulose membrane, and the sample pad is tightly pressed against one end, far away from the nitrocellulose membrane, of the bonding pad; and a detection line and a quality control line are sequentially arranged on the nitrocellulose membrane from one end close to the bonding pad to one end far away from the bonding pad. The immunofluorescence chromatography test paper can be applied to the rapid detection of etomidate, and has strong detection specificity and high sensitivity. The invention provides powerful technical support for on-site rapid detection work and high-throughput screening of etomidate.
Description
Technical Field
The invention relates to the technical field of qualitative identification and drug-arresting detection, in particular to etomidate immunofluorescence chromatography rapid detection test paper and a detection method.
Background
At present, the main analysis and detection means of etomidate comprise High Performance Liquid Chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS/MS), portable mass spectrometry and other analysis methods. The existing analysis method has advantages in qualitative and quantitative detection of in-vivo and in-vitro detection materials, but cannot completely meet the requirements of on-site rapid detection and high-throughput screening due to limitations of the existing analysis method in terms of instrument condition requirements, analysis time, portability and the like. Further development and enrichment of rapid detection means for etomidate is needed.
The immunochromatography diagnosis method based on the chromatography technology and antigen-antibody specific immune reaction is a dominant product in the field of rapid detection. The immunochromatography technology is a lateral flow rapid detection technology for detecting the content of a substance to be detected in a sample by using a large-aperture microporous filter membrane as a carrier and a solid-phase marker according to an immunological principle. The invention aims to develop an etomidate immunofluorescence chromatography rapid detection paper card, thereby providing technical support for on-site rapid detection work and high-throughput screening of etomidate.
Disclosure of Invention
The invention aims to provide etomidate immunofluorescence chromatography rapid detection test paper and a detection method, which are used for solving the problems in the prior art. The immunofluorescence chromatography test paper can be applied to the rapid detection of etomidate, and has strong detection specificity and high sensitivity, and the detection limit can reach 20 ng/mL. The invention provides powerful technical support for on-site rapid detection work and high-throughput screening of etomidate.
In order to achieve the above object, the present invention provides the following solutions:
The invention provides etomidate immunofluorescence chromatography rapid test paper which comprises a bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad, wherein the bottom plate is provided with a sample pad; the nitrocellulose membrane is attached to the bottom plate, the water absorbing pad and the bonding pad are respectively tightly pressed against two ends of the nitrocellulose membrane, and the sample pad is tightly pressed against one end, far away from the nitrocellulose membrane, of the bonding pad;
The nitrocellulose membrane is sequentially provided with a detection line and a quality control line from one end close to the bonding pad to one end far away from the bonding pad;
The detection line is coated with antigen; the quality control line is coated with goat anti-mouse secondary antibody;
The binding pad carries a fluorescent quantum dot-monoclonal antibody complex;
The fluorescent quantum dot-monoclonal antibody compound is obtained by coupling fluorescent quantum dots and etomidate monoclonal antibodies.
Further, the bottom plate is a PVC plate.
Further, the nitrocellulose membrane is a Millipore 135s membrane.
Further, the fluorescent quantum dots are water-soluble CdTe/CdS/ZnS quantum dots.
Further, the sample pad is obtained by soaking a glass fiber membrane or a polyester fiber membrane in a Tris-HCL buffer system containing 2.0% Tween 20, and then drying;
The bonding pad is obtained by soaking a glass fiber film or a polyester fiber film in a Tris-HCL buffer system containing 2.0% Tween 20, drying and spraying; the spraying treatment refers to spraying the fluorescent quantum dot-monoclonal antibody compound.
Further, when the detection line is coated with the antigen, the concentration of the coated antigen solution used is 1.5 mg/mL.
Further, when the quality control line coats the goat anti-mouse secondary antibody, the goat anti-mouse secondary antibody solution is obtained by 20000 times dilution of the goat anti-mouse secondary antibody.
The invention also provides application of the etomidate immunofluorescence chromatography rapid detection test paper in etomidate detection for non-disease diagnosis purposes.
The invention also provides a method for detecting etomidate in a liquid sample for non-disease diagnosis purpose, which comprises the following steps:
dropwise adding a liquid sample to be detected into a sample pad of the etomidate immunofluorescence chromatography rapid detection test paper, observing a fluorescence signal after chromatography treatment, and judging according to an observation result as follows:
If fluorescent strips appear on the quality control line and the detection line, judging that etomidate is not detected or exceeds a detection limit;
If the quality control line has a fluorescent strip and the detection line has no fluorescent strip, judging that etomidate is detected;
if the quality control line has no fluorescent strip, judging whether the detection line has the fluorescent strip or not, and judging that the detection result is invalid.
The invention also provides a method for detecting etomidate in a solid sample for non-disease diagnosis purpose, which comprises the following steps:
after grinding the solid sample to be detected, dissolving the solid sample to be detected by using an aqueous solution containing an organic solvent to obtain a liquid sample to be detected;
Dropwise adding the liquid sample to be detected into a sample pad of the etomidate immunofluorescence chromatography rapid detection test paper, observing a fluorescence signal after chromatography treatment, and judging according to the observation result as follows:
If fluorescent strips appear on the quality control line and the detection line, judging that etomidate is not detected;
If the quality control line has a fluorescent strip and the detection line has no fluorescent strip, judging that etomidate is detected;
if the quality control line has no fluorescent strip, judging whether the detection line has the fluorescent strip or not, and judging that the detection result is invalid.
The invention discloses the following technical effects:
The immunofluorescence chromatography test paper capable of being used for quick detection of etomidate is constructed through serial works such as fluorescent marker coupling, membrane material screening, buffer system optimization, surfactant optimization, streak dosage screening, methodology investigation and the like. The immunofluorescence chromatography test paper can be applied to the rapid detection of etomidate, and has strong detection specificity and high sensitivity, and the detection limit can reach 20 ng/mL. The invention provides powerful technical support for on-site rapid detection work and high-throughput screening of etomidate.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic structural diagram of etomidate immunofluorescence chromatography test paper of the invention; wherein, 1-a bottom plate; 2-sample pad; 3-a bonding pad; 4-nitrocellulose membrane; 5-detecting lines; 6-a quality control line; 7-water absorbing pad.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, 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. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Example 1
As shown in figure 1, the invention constructs an etomidate immunofluorescence chromatography test paper, which comprises a bottom plate (1), a sample pad (2), a binding pad (3), a Nitrocellulose (NC) membrane (4) and a water absorption pad (7); the nitrocellulose membrane is attached to the bottom plate, the water absorption pad and the bonding pad are respectively tightly pressed against two ends of the nitrocellulose membrane, and the sample pad is tightly pressed against one end of the bonding pad, which is far away from the nitrocellulose membrane;
A detection line (5, namely a T line) and a quality control line (6, namely a C line) are sequentially arranged on the nitrocellulose membrane from one end close to the bonding pad to one end far from the bonding pad; the detection line is coated with antigen; coating a quality control line with goat anti-mouse secondary antibody;
the conjugate pad carries a fluorescent quantum dot-mab complex.
The absorbent pad length, sample pad, conjugate pad and NC membrane were all 0.35 cm in width, and 2. 2 cm, 1.8 cm, 0.5 cm and 3 cm in length, respectively. The preparation process of the etomidate immunofluorescence chromatography test paper is as follows:
1. preparation of fluorescent quantum dot labeled antibody
Water-soluble CdTe/CdS/ZnS fluorescent quantum dots (from Beijing North Dacron technologies Co., ltd., product No. W-511-620) were formulated into a 1. Mu.M fluorescent quantum dot solution using sodium borate buffer (pH=7.4), and etomidate monoclonal antibodies (from Beijing Huakang Toyoshiba Biotechnology Co., ltd.) were formulated into 4 mg/mL etomidate monoclonal antibody solutions using the same sodium borate buffer. Carbodiimide (EDC) and N-hydroxysulfosuccinimide (NHS) were dissolved in 10mM sodium borate buffer, respectively, to prepare 2: 2 mg/mL EDC solution and 1:1 mg/mL NHS solution, respectively. 500. Mu.L of EDC solution and 500. Mu.L of fluorescent quantum dot solution were mixed and stirred at low speed (rotation speed 100 rpm) at room temperature for 30: 30 min. After completion, 500. Mu.L of NHS solution was added thereto, followed by stirring at room temperature for 30min at low speed, then 500. Mu.L of etomidate monoclonal antibody solution was added thereto, and stirring at room temperature for 2: 2h at low speed was continued to obtain a mixture. The resulting mixture was purified using an ultrafiltration tube (MWCO: 10 kDa) and concentrated to 50. Mu.L, and centrifuged at 6000 rpm for 10 min to obtain a precipitate, which was the fluorescent quantum dot-mab complex.
2. Pretreatment of sample pad and conjugate pad
In the immunochromatography test paper card preparation process, a buffer solution is required to be used for pretreatment of a binding pad and a sample pad. In the research and development process, three different buffer solutions (Tris-HCL, phosphate buffer solution and boric acid buffer solution) are optimally screened, and an optimal buffer system is screened according to the color development condition and the background intensity of the T/C line in the detection result. Results of testing etomidate-containing urine, hair and blood samples using different buffer solutions are shown in table 1. The result shows that the interference of different pH matrixes on the detection result is minimum when Tris-HCL is used as a buffer system in consideration of the application scene of the test paper card and the complexity of a sample to be detected.
Table 1 results of testing urine, hair and blood samples using different buffer solutions
The surfactant has the main functions of improving hydrophilicity and wettability, reducing the surface tension of liquid, and the addition of the surfactant in the chromatographic system is favorable for the rapid and full reaction of antigen and antibody. In order to optimize and ensure the detection result, the dosage of the surfactant Tween20 used for preparing the test paper card is screened. After different doses (volume fractions of 0.5%, 1.0%, 2.0% and 3.0%) of Tween20 were added to the buffer system, respectively, the effect of the different doses of surfactant on the release of the Quantum dot labeled etomidate antibody and the T/C line fluorescence development was tested. According to the screening results of Table 2, the requirements of avoiding background residues, clear fluorescent stripes and no interference of T/C lines are comprehensively considered, and the addition amount of the surfactant is finally determined to be 2.0% Tween 20.
TABLE 2 results of etomidate-containing samples tested with different doses of Tween 20
The glass fiber film sample pad (or the polyester fiber film sample pad) and the glass fiber film bonding pad (or the polyester fiber film bonding pad) are respectively soaked in a Tris-HCL buffer system containing 2.0% Tween 20 for 5 s, taken out, dried and stored. And uniformly spraying the prepared fluorescent quantum dot-monoclonal antibody compound on the pretreated bonding pad by using a metal spraying instrument, wherein the spraying amount is 5 mu L/cm, and drying and preserving.
3. NC film scribing
NC membrane is the binding position of etomidate antibody and antigen on the test paper, and is an important component of the test paper. Two NC membrane materials, millipore 135s and Millipore 180s, were examined, and optimized and selected based on their effect on chromatographic procedures and detection. Tests have found that due to the difference in density, the chromatography speed of Millipore 180s is relatively slow, and the chromatography time of Millipore 135s is faster than that of Millipore 180s, which means that the reaction time is longer when a Millipore 180s system is used and shorter when a Millipore 135s system is used. According to the chemical structure and molecular weight (C 14H16N2O2, 244.3 Da) of etomidate and the competition method adopted by the test paper detection system, the short antigen and antibody binding reaction time is more beneficial to guaranteeing the sensitivity of the test paper, so that Millipore 135s is finally selected as NC membrane material in the research and development of the quick test paper.
And screening and optimizing the scribing dose of the C line and the T line. Respectively selecting mass ratio 1: 10000. 1: 20000. 1: membrane streaking was performed at three concentrations of C-line goat anti-mouse secondary antibodies diluted with 50000 and three T-line antigen concentrations of 1.0, 1.5, 2.0 mg/mL. Tests were performed using negative sample solutions and the effect of different dose combinations on the test results of the test strips was compared. The lowest concentration of the T/C line fluorescent strip which is clear and has no background interference is selected as the scribing concentration of the T line and the C line. And finally determining that the streak concentration of the anti-mouse secondary antibody of the C line sheep is 1:20000 and the streak concentration of the T line antigen is 1.5 mg/mL.
The NC film scribing operation was determined as follows, through material and dose optimization:
The dialyzed coating antigen (diluted 1:20000) and goat anti-mouse secondary antibody (concentration 1.5 mg/mL, goat anti-mouse secondary antibody is purchased from Beijing pride gene technology Co., ltd.) are respectively loaded by using a metal spraying and film drawing instrument, and the goat anti-mouse secondary antibody with the width of 1mm is uniformly sprayed on the water absorbing end of the Millipore 135s NC film to form a quality control line (C line). And uniformly spraying a coating antigen with the width of 1mm on the sample end of the NC film to form a detection line (T line), and drying at room temperature.
4. Assembly of test paper
Firstly, pasting the NC film after scribing on a PVC board (one end is a sample end, and the other end is a water absorption end), and controlling the distance between the NC film and the edge of the water absorption end of the PVC board to be 2 cm. And (3) attaching the pretreated bonding pad above the NC film, and controlling the overlapping length of the bonding pad and the NC film to be 0.1 cm. The pretreated sample pad is stuck on a PVC plate below the bonding pad, and the overlapping length of the sample pad and the bonding pad is controlled to be 2 mm. The water absorption pad is stuck on a PVC plate below the NC film, and the overlapping of the water absorption pad and the NC film is controlled to be 2 mm. And compacting the assembled test paper by adopting a batch lamination system, cutting the test paper into a width of 0.35 cm, sealing the test paper in an aluminum foil bag containing a drying agent, and storing the test paper in a dark place for later use.
And (3) carrying out methodology investigation, diluting etomidate into a series of concentration gradient solutions, respectively and simultaneously sucking 50 mu L of the solutions into a sample adding hole, repeating each concentration test for three times in parallel, observing the result under 365 nm ultraviolet light excitation after 5 minutes, and finally determining that the detection limit of the test strip is 20 ng/mL. When the etomidate concentration in the sample to be detected is more than 20 ng/mL, the sample to be detected fully competes and binds with the fluorescent-labeled monoclonal antibody sprayed on the membrane, so that the antibody is inhibited from being bound with the antigen sprayed on the T detection line along with the chromatographic system, the T detection line does not show a fluorescent strip, and the result is judged to be positive. When the etomidate concentration in the sample to be detected is less than 20 ng/mL, the etomidate in the sample to be detected is insufficient to be combined with the fluorescent marked etomidate monoclonal antibody in the sample pad in a fully competitive manner, so that part of the antibody is combined with the antigen sprayed at the T line after being unfolded along with a chromatographic system, and finally two fluorescent strips are displayed at the T line and the C line, thereby judging that the result is negative.
Example 2
In order to examine the specificity of the etomidate immunofluorescence chromatography test paper and the rapid detection method constructed by the invention, 20 compounds of the amphetamine (methamphetamine, amphetamine and methylenedioxymethamphetamine), the zolam (triazolam, midazolamne and esmolam), the synthetic cannabinoids (4F-MDMB-BUTINACA and 5F-APINAC), the synthetic carbostyril (carboximide and mecarbazepine), the zepam (fluazepam, diazepam and lorazepam), the methadone, caffeine, fentanyl, ketamine and non-shell pharmaceutical compositions (reserpine, ursolic acid and metformin) are used for detection, and whether cross reaction occurs or not, namely false positive is observed. The detection results are shown in Table 3, and the results show that the test paper is negative to the detection results of 20 selected compounds except etomidate, and has no cross reaction, so that the etomidate immunochromatography test paper card and the quick detection method have higher specificity.
TABLE 3 results of specificity experiments
Example 3
Qualitative detection of etomidate in urine:
The urine sample to be tested is balanced to room temperature, 2 drops of the test paper prepared in the embodiment 1 are dripped into the sample adding place, and the test paper is tested under a fluorescence excitation light source (365 nm) after waiting for 10min (during which the sample solution migrates to the other end of the test paper due to the capillary action driven by the absorbent paper). Observing fluorescent signals, and if fluorescent strips appear on both the quality control line and the detection line, judging that the detection sample is negative, namely, etomidate is not detected (or the detection limit is exceeded); if the quality control line has a fluorescent strip and the detection line has no fluorescent strip, judging that the detection sample is positive, namely detecting etomidate; if the quality control line has no fluorescent strip, judging whether the detection line has the fluorescent strip or not, and judging that the detection result is invalid.
Example 4
Qualitative detection of etomidate in hair:
30 mg hair samples were weighed and 1mL volume percent 5% aqueous methanol was added and milled using a high speed mill. The grinding fluid is balanced to room temperature, the detection test paper prepared in the embodiment 1 is dripped into a sample adding hole for 2 drops, and the detection is carried out under a fluorescence excitation light source (365 nm) after waiting for 10min. Observing fluorescent signals, and if fluorescent strips appear on both the quality control line and the detection line, judging that the detection sample is negative, namely, etomidate is not detected (or the detection limit is exceeded); if the quality control line has a fluorescent strip and the detection line has no fluorescent strip, judging that the detection sample is positive, namely detecting etomidate; if the quality control line has no fluorescent strip, judging whether the detection line has the fluorescent strip or not, and judging that the detection result is invalid.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (10)
1. An etomidate immunofluorescence chromatography rapid test paper is characterized by comprising a bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad; the nitrocellulose membrane is attached to the bottom plate, the water absorbing pad and the bonding pad are respectively tightly pressed against two ends of the nitrocellulose membrane, and the sample pad is tightly pressed against one end, far away from the nitrocellulose membrane, of the bonding pad;
The nitrocellulose membrane is sequentially provided with a detection line and a quality control line from one end close to the bonding pad to one end far away from the bonding pad;
The detection line is coated with antigen; the quality control line is coated with goat anti-mouse secondary antibody;
The binding pad carries a fluorescent quantum dot-monoclonal antibody complex;
The fluorescent quantum dot-monoclonal antibody compound is obtained by coupling fluorescent quantum dots and etomidate monoclonal antibodies.
2. An etomidate immunofluorescence chromatography rapid test strip according to claim 1, wherein said base plate is a PVC plate.
3. An etomidate immunofluorescence chromatography rapid test strip according to claim 1, wherein said nitrocellulose membrane is a Millipore 135s membrane.
4. An etomidate immunofluorescence chromatography rapid test paper according to claim 1, wherein the fluorescent quantum dots are water-soluble CdTe/CdS/ZnS quantum dots.
5. An etomidate immunofluorescence chromatography rapid test paper according to claim 1, wherein the sample pad is obtained by soaking a glass fiber membrane or a polyester fiber membrane in a Tris-HCL buffer system containing 2.0% Tween 20, and drying;
The bonding pad is obtained by soaking a glass fiber film or a polyester fiber film in a Tris-HCL buffer system containing 2.0% Tween 20, drying and spraying; the spraying treatment refers to spraying the fluorescent quantum dot-monoclonal antibody compound.
6. An etomidate immunofluorescence chromatography rapid test strip according to claim 1, wherein when said detection line is coated with said antigen, the concentration of the coated antigen solution used is 1.5 mg/mL.
7. An etomidate immunofluorescence chromatography rapid test paper according to claim 1, wherein when the quality control line is coated with the goat anti-mouse secondary antibody, the goat anti-mouse secondary antibody solution is obtained by 20000 times dilution of the goat anti-mouse secondary antibody.
8. Use of an etomidate immunofluorescence chromatography rapid test strip according to any one of claims 1-7 for detection of etomidate for non-disease diagnostic purposes.
9. A method for detecting etomidate in a liquid sample for non-disease diagnostic purposes, comprising the steps of:
Dropping a liquid sample to be detected into a sample pad of the etomidate immunofluorescence chromatography rapid detection test paper according to any one of claims 1-7, observing a fluorescence signal after chromatography treatment, and judging according to the observation result as follows:
If fluorescent strips appear on the quality control line and the detection line, judging that etomidate is not detected or exceeds a detection limit;
If the quality control line has a fluorescent strip and the detection line has no fluorescent strip, judging that etomidate is detected;
if the quality control line has no fluorescent strip, judging whether the detection line has the fluorescent strip or not, and judging that the detection result is invalid.
10. A method for detecting etomidate in a solid sample for non-disease diagnostic purposes, comprising the steps of:
after grinding the solid sample to be detected, dissolving the solid sample to be detected by using an aqueous solution containing an organic solvent to obtain a liquid sample to be detected;
dropping the liquid sample to be detected into the sample pad of the etomidate immunofluorescence chromatography rapid detection test paper according to any one of claims 1-7, observing a fluorescence signal after chromatography treatment, and judging according to the observation result as follows:
If fluorescent strips appear on the quality control line and the detection line, judging that etomidate is not detected;
If the quality control line has a fluorescent strip and the detection line has no fluorescent strip, judging that etomidate is detected;
if the quality control line has no fluorescent strip, judging whether the detection line has the fluorescent strip or not, and judging that the detection result is invalid.
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