CN116794305A - Quick detection kit for swab samples and preparation method thereof - Google Patents
Quick detection kit for swab samples and preparation method thereof Download PDFInfo
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- CN116794305A CN116794305A CN202310886540.3A CN202310886540A CN116794305A CN 116794305 A CN116794305 A CN 116794305A CN 202310886540 A CN202310886540 A CN 202310886540A CN 116794305 A CN116794305 A CN 116794305A
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
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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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G01N33/54387—Immunochromatographic test strips
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
Abstract
The application discloses a rapid detection kit for a swab sample. The rapid detection kit for the swab sample comprises a kit main body, a test strip and a swab component, wherein the test strip and the swab component are both arranged in the kit main body, the swab component is provided with a detection channel, one end of the detection channel extends to the sample loading end of the test strip, the sample loading end of the test strip is at least partially positioned in the detection channel, and the other end of the detection channel faces the outside of the kit main body so as to be used for inserting a swab sample into the sample loading end of the contact test strip. Above-mentioned quick detection kit of swab sample combines together test strip and swab sample, and the swab sample need not elution treatment and directly contacts with the test strip and realize detecting, novel structure can realize the direct application of sample detection integration of swab, and detection efficiency is high, has solved the technical problem that the conventional art detects many and easily appear polluting of step.
Description
Technical Field
The application relates to the technical field of biological detection, in particular to a rapid detection kit for a swab sample and a preparation method thereof.
Background
In the development process of medical clinical detection technology, gold-labeled monoclonal antibody rapid detection test paper is rapidly developed, the detection technology can detect more than 100 antigen substances, including various hormones, cancer antigens, viruses, bacteria, antibodies, synthetic antigens, drug abuse and the like, and the gold-labeled monoclonal antibody rapid detection test paper has been applied to a plurality of clinical detection projects such as various infectious diseases, early cancers, venereal diseases, AIDS, early pregnancy, ovulation, fetal malformation early detection and the like. The method has the advantages incomparable with the common conventional method in various aspects such as sample requirement, detection time, instrument and equipment, personnel requirement, test cost, safety and the like of clinical measurement, and can rapidly, simply, conveniently and intuitively detect a plurality of detection results which can be obtained in hospitals only with time and labor consumption in about 10 minutes with extremely low cost. Test paper products based on gold-labeled monoclonal antibody rapid detection technology have market potential of about 8000 ten thousand per year only for female reproductive health (such as eugenic, contraceptive and infertility) detection. However, the rapid detection technology of the gold-labeled monoclonal antibody is limited by the color development sensitivity, the sensitivity can only reach 1ng/mL, and the rapid detection technology of the gold-labeled monoclonal antibody can not meet the requirements of clinical disease diagnosis, especially the early diagnosis of serious diseases due to the limitations of the sensitivity and the difficulty in quantification of the gold-labeled method.
The colloidal gold immunochromatography technology is an immunochromatography detection method using colloidal gold nanoparticles as an immune marker, and the result is intuitive to read and judge without any instrument and equipment, so that the colloidal gold immunochromatography technology is widely focused. Swab samples (including nasal swabs, pharyngeal swabs, and other swab samples) are in great demand during rapid testing. In the rapid diagnosis and use process of the swab sample in the prior art, the detection card is independently separated from the sample extraction, the swab sample needs to be treated independently, the operation steps are more, and pollution is very easy to occur.
Disclosure of Invention
Based on this, an embodiment of the present application provides a rapid detection kit for a swab sample. The rapid detection kit for the swab sample can realize rapid and accurate color development so as to detect an object to be detected in the swab sample.
The embodiment of the application provides a rapid detection kit for a swab sample.
The utility model provides a quick detection kit of swab sample, includes kit main part, test paper strip and swab subassembly, the test paper strip with the swab subassembly all install in the kit main part, the swab subassembly has detection channel, detection channel's one end extends to the loading end of test paper strip, the loading end of test paper strip is at least partly located in the detection channel, detection channel's the other end is towards the outside of kit main part for the swab sample to insert to the contact the loading end of test paper strip.
In some embodiments, a detection window is provided on the kit main body, and the detection window is opposite to the detection line and the control line on the test strip.
In some embodiments, the kit main body comprises a first shell and a second shell, the first shell is connected with the second shell and encloses a detection cavity, a detection station communicated with the detection cavity is arranged on the first shell or the second shell, the test strip and the swab component are both installed in the detection cavity, and the swab component is located at the detection station.
In some embodiments, the first housing is provided with the detection station facing the inner surface of the detection chamber, the detection station is used for mounting the swab component, and the second housing is provided with a mounting station used for mounting the test strip.
In some of these embodiments, the first housing is removably coupled to the second housing.
In some embodiments, the kit body further comprises a connection assembly, at least one of the connection assemblies is connected to the first housing and/or the second housing, and the first housing and the second housing are connected by the connection assembly.
In some of these embodiments, the test strip is mounted to the second housing via the connection assembly, and the swab assembly is mounted to the first housing via the connection assembly.
In some embodiments, the connecting component is one or more of double faced adhesive tape, a magnetic alignment adsorption structure and a rivet fastening structure.
In some embodiments, the swab assembly is provided with a pressing mechanism for fixing a sample swab, the pressing mechanism is provided with a first positioning groove for fixing a swab head of the sample swab and a second positioning groove for fixing a swab rod of the sample swab, the first positioning groove and the second positioning groove form the detection channel, and at least part of a sample loading end of the test strip is located in the first positioning groove.
In some of these embodiments, the swab sample rapid detection kit further comprises a swab sample comprising a swab head for sampling and a swab stem connected to the swab head.
In some of these embodiments, the swab sample comprises one or more of a nasal swab sample, a pharyngeal swab sample, a nasopharyngeal swab, a urethral swab, and a cervical swab sample.
In some embodiments, the preparation material of the kit main body is selected from one or more of hard paper, polyethylene terephthalate, polymethyl methacrylate and polypropylene.
In some embodiments, the swab component is made from a material selected from one or more of polyvinyl chloride, polyethylene terephthalate, polyethylene, polypropylene, polyvinylidene chloride, and polycarbonate.
The embodiment of the application also provides a preparation method of the rapid detection kit for the swab sample,
a preparation method of a swab sample rapid detection kit comprises the following steps:
(1) Preparing a test strip;
(2) The test strip is arranged at a preset position in the main body of the kit, the swab component is arranged at the preset position in the main body of the kit, one end of a detection channel of the swab component extends to the loading end of the test strip, so that the loading end of the test strip is at least partially positioned in the detection channel, and the other end of the detection channel faces the outside of the main body of the kit so as to be used for inserting a swab sample to be contacted with the loading end of the test strip.
In some embodiments, the test strip preparation method comprises the following steps:
preparing a conjugate pad:
preparing a colloidal gold nanoparticle solution;
preparing a colloidal gold virus antibody complex by adopting the colloidal gold nanoparticle solution, diluting the colloidal gold virus antibody complex and the colloidal gold quality control antibody complex by using a gold-labeled diluent to prepare a conjugate treatment solution, and coating the conjugate treatment solution on a conjugate pad;
preparation of nitrocellulose film: respectively diluting a virus antigen capture antibody and a quality control capture antibody with coating liquid, then spot-coating the diluted virus antigen capture antibody and the quality control capture antibody on corresponding positions of a nitrocellulose membrane, and drying and fixing the diluted virus antigen capture antibody and the quality control capture antibody to obtain the nitrocellulose membrane;
preparing a sample pad: smearing the treatment liquid on a sample pad, and drying for later use;
and sequentially adhering the sample pad, the conjugate pad, the nitrocellulose membrane and the absorbent paper on a bottom plate, and cutting into test strips with preset sizes.
In some embodiments, the volume ratio of the colloidal gold virus antibody complex in the conjugate treatment solution is 10% -20%, and the volume ratio of the colloidal gold quality control antibody complex is 2% -5%;
and/or the virus antigen capture antibody and the quality control capture antibody are respectively diluted to 0.5-2 mg/mL and 0.5-2 mg/mL by the coating liquid.
Above-mentioned quick detection kit of swab sample combines together test strip and swab sample, and the swab sample need not elution treatment and directly contacts with the test strip and realize detecting, novel structure can realize the direct application of sample detection integration of swab, and detection efficiency is high, has solved the technical problem that the conventional art detects many and easily appear polluting of step. According to the rapid detection kit for the swab samples, the test paper strip and the card are combined and assembled to form the reagent with the detection function, and the swab device is arranged at the sample adding end, so that the sample can be directly added for detection after sampling the swab, the operation steps are shortened, and the efficiency is improved, and the detection card for the swab samples is prepared, so that the integration of the direct sample adding and detection of the swab is realized, the operation is friendly, the sample is not easy to pollute, the detection efficiency is high, and the detection accuracy is high.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings that are required to be used in the description of the embodiments will be briefly described below. It is evident that the figures in the following description are only some embodiments of the application, from which other figures can be obtained without inventive effort for a person skilled in the art.
For a more complete understanding of the present application and the advantages thereof, reference is now made to the following descriptions taken in conjunction with the accompanying drawings. Wherein like reference numerals refer to like parts throughout the following description.
FIG. 1 is a schematic diagram of a rapid detection kit for a swab sample according to one embodiment of the present application;
FIG. 2 is a schematic diagram of a rapid detection kit for a swab sample according to one embodiment of the present application;
FIG. 3 is a schematic diagram of a first housing and a second housing of a rapid swab sample detection kit according to one embodiment of the present application;
FIG. 4 is a schematic diagram of a swab assembly of a rapid swab sample detection kit according to one embodiment of the present application;
fig. 5 is a schematic diagram illustrating a rapid detection kit for a swab sample according to an embodiment of the present application.
Description of the reference numerals
10. A swab sample rapid detection kit; 100. a kit body; 101. a first housing; 1011. a detection window; 1012. detecting a station; 102. a second housing; 103. a connection assembly; 200. a test strip; 300. a swab assembly; 301. a pressing and holding mechanism; 3011. a first positioning groove; 3012. a second positioning groove; 400. a swab sample.
Detailed Description
In order that the above objects, features and advantages of the application will be readily understood, a more particular description of the application will be rendered by reference to the appended drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. The present application may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the application, whereby the application is not limited to the specific embodiments disclosed below.
In the description of the present application, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present application and simplifying the description, and do not indicate or imply that the device or element being referred to must have a specific orientation, be configured and operated in a specific orientation, and therefore should not be construed as limiting the present application.
In the present application, unless explicitly specified and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly, through intermediaries, or both, may be in communication with each other or in interaction with each other, unless expressly defined otherwise. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art according to the specific circumstances.
In the present application, unless expressly stated or limited otherwise, a first feature "up" or "down" a second feature may be the first and second features in direct contact, or the first and second features in indirect contact via an intervening medium. Moreover, a first feature being "above," "over" and "on" a second feature may be a first feature being directly above or obliquely above the second feature, or simply indicating that the first feature is level higher than the second feature. The first feature being "under", "below" and "beneath" the second feature may be the first feature being directly under or obliquely below the second feature, or simply indicating that the first feature is less level than the second feature.
In the description of the present application, the meaning of a number is one or more, the meaning of a number is two or more, and greater than, less than, exceeding, etc. are understood to exclude the present number, and the meaning of a number is understood to include the present number. The description of the first and second is for the purpose of distinguishing between technical features only and should not be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of the technical features indicated.
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 application belongs. The terminology used herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The embodiment of the application provides a rapid detection kit 10 for a swab sample, which aims to solve the problems of more detection steps and easy pollution in the traditional technology. The rapid swab sample detection kit 10 will be described with reference to the accompanying drawings.
Referring to fig. 1, fig. 1 is a schematic structural diagram of a swab sample rapid detection kit 10 according to an embodiment of the present application. The rapid detection kit 10 for the swab sample can be used for detecting biological samples, for example, the rapid detection kit 10 for the swab sample can be used for rapidly detecting the swab sample 400, and has high detection efficiency and accurate detection result.
In order to more clearly illustrate the structure of the rapid swab sample detection kit 10, the rapid swab sample detection kit 10 will be described with reference to the accompanying drawings.
Referring to fig. 1, a rapid test kit 10 for a swab sample includes a kit body 100, a test strip 200, and a swab assembly 300.
The test strip 200 and the swab assembly 300 are both mounted in the kit body 100. The swab assembly 300 has a detection channel. One end of the detection channel extends to the loading end of the test strip 200. The loading end of the test strip 200 is at least partially disposed within the detection channel. The other end of the detection channel faces the outside of the cartridge body 100 for insertion of the swab sample 400 into the loading end of the contact test strip 200.
Above-mentioned quick detection kit of swab sample 10 combines together test strip 200 and swab sample 400, and swab sample 400 need not elution treatment and directly contacts with test strip 200 and realize detecting, novel structure can realize the direct application of sample detection integration of swab, and detection efficiency is high, has solved the technical problem that the conventional art detects many and easily appear polluting of step.
In some of these embodiments, a detection window 1011 is provided on the cartridge body 100. The detection window 1011 is opposite to the detection line and the control line on the test strip 200.
In some of these embodiments, the kit body 100 includes a first housing 101 and a second housing 102. The first housing 101 is connected to the second housing 102 and encloses a detection chamber. The first housing 101 or the second housing 102 is provided with a detection station 1012 communicating with the detection chamber. The test strip 200 and the swab assembly 300 are both mounted in the detection chamber. The swab assembly 300 is located at the inspection station 1012.
In some of these embodiments, the first housing 101 is provided with a detection station 1012 on an inner surface facing the detection chamber, the detection station 1012 being configured for mounting the swab assembly 300, and the second housing 102 is provided with a mounting station configured for mounting the test strip 200.
In some of these embodiments, the first housing 101 is detachably connected to the second housing 102.
In some embodiments, referring to fig. 2, fig. 2 is a schematic diagram of a rapid swab sample detection kit 10 according to an embodiment of the present application, the kit main body 100 further includes a connection component 103, at least one connection component 103 is connected to the first housing 101 and/or the second housing 102, and the first housing 101 and the second housing 102 are connected by the connection component 103.
In some of these embodiments, the test strip 200 is mounted to the second housing 102 via the connection assembly 103, and the swab assembly 300 is mounted to the first housing 101 via the connection assembly 103.
In some embodiments, the connection component 103 is one or more of double sided tape, magnetic alignment suction structure, and rivet fastening structure. Preferably, in one specific example, the connection assembly 103 is double sided tape.
In some embodiments, referring to fig. 4, fig. 4 is a schematic diagram of a swab assembly 300 of a rapid detection kit for a swab sample 10 according to an embodiment of the present application, a pressing mechanism 301 for fixing a sample swab is disposed on the swab assembly 300, the pressing mechanism 301 has a first positioning groove 3011 for fixing a swab head of the sample swab and a second positioning groove 3012 for fixing a swab rod of the swab sample 400, the first positioning groove 3011 and the second positioning groove 3012 form a detection channel, and at least a portion of a loading end of the test strip 200 is located in the first positioning groove 3011.
In some embodiments, referring to fig. 4, the first positioning groove 3011 of the swab assembly 300 is an oval groove, and can be used for the swab head to clamp, the second positioning groove 3012 is an elongated groove, the second positioning groove 3012 is convenient for rotating the swab rod, and the swab assembly 300 is used as a swab collecting fixing and loading function.
In some embodiments, the detection window 1011 and the detection station 1012 are both arranged on the first casing 101, so that a detection person can directly and simultaneously observe the insertion and detection results of the swab sample 400, and the detection window is convenient and visual.
In some embodiments, referring to fig. 3, fig. 3 is a schematic diagram of a first housing 101 and a second housing 102 of a rapid detection kit for a swab sample 10 according to one embodiment of the present application, and a detection station 1012 is a notch structure.
In some embodiments, referring to fig. 5, fig. 5 is a schematic diagram illustrating the cooperation of the rapid detection kit for a swab sample 10 and the swab sample 400 according to an embodiment of the present application, the rapid detection kit for a swab sample 10 further includes the swab sample 400, and the swab sample 400 includes a swab head and a swab rod connected to the swab head, and the swab head is used for sampling.
In some of these embodiments, swab sample 400 includes one or more of a nasal swab sample, a pharyngeal swab sample, a nasopharyngeal swab, a urethral swab, and a cervical swab sample. The swab sample 400 has good handling and transplanting effects.
In some of these embodiments, the preparation material of the kit body 100 is selected from one or more of hard paper, polyethylene terephthalate (PET), polymethyl methacrylate (PMMA), and polypropylene (PP). Specifically, the first casing 101 and the second casing 102 are both made of hard paper, and after the test strip 200 and the swab component 300 are placed between the first casing 101 and the second casing 102 at corresponding positions, the connection component 103 such as double faced adhesive tape is arranged, and then press-fit treatment is performed.
In some of these embodiments, the swab component 300 is made from a material selected from one or more of polyvinyl chloride (PVC), polyethylene terephthalate (PET), polyethylene terephthalate (PETP), polyethylene (PE), polypropylene (PP), polyvinylidene chloride (PVDC), and Polycarbonate (PC).
An embodiment of the present application also provides a method for preparing the rapid test kit 10 for swab samples,
(1) Preparing a test strip 200;
(2) The test strip 200 is mounted at a predetermined position in the kit body 100, the swab assembly 300 is mounted at a predetermined position in the kit body 100, one end of the detection channel of the swab assembly 300 extends to the loading end of the test strip 200, so that the loading end of the test strip 200 is at least partially positioned in the detection channel, and the other end of the detection channel faces the outside of the kit body 100 for inserting the swab sample 400 into the loading end of the contact test strip 200.
In some embodiments, the method for preparing the test strip 200 includes:
preparing a conjugate pad: preparing a colloidal gold nanoparticle solution; preparing a colloidal gold virus antibody complex by using a colloidal gold nanoparticle solution; diluting the colloidal gold virus antibody complex and the colloidal gold quality control antibody complex by using a gold-labeled diluent to prepare a conjugate treatment solution, and coating the conjugate treatment solution on a conjugate pad;
preparation of nitrocellulose film: respectively diluting a virus antigen capture antibody and a quality control capture antibody with coating liquid, then spot-coating the diluted virus antigen capture antibody and the quality control capture antibody on corresponding positions of a nitrocellulose membrane, and drying and fixing the diluted virus antigen capture antibody and the quality control capture antibody to obtain the nitrocellulose membrane;
preparing a sample pad: smearing the treatment liquid on a sample pad, and drying for later use;
the sample pad, conjugate pad, nitrocellulose membrane, and absorbent paper are sequentially adhered to the base plate, and cut into test strips 200 of a predetermined size.
In some embodiments, the volume ratio of the colloidal gold virus antibody complex in the conjugate treatment solution is 10% -20%, and the volume ratio of the colloidal gold quality control antibody complex is 2% -5%.
In some embodiments, the virus antigen capture antibody and the quality control capture antibody are diluted to 0.5-2 mg/mL and 0.5-2 mg/mL, respectively, with a coating solution. Wherein, the coating liquid can be nitrocellulose membrane coating liquid.
In some of these embodiments, the drying immobilization conditions in preparing the nitrocellulose membrane are: drying and fixing at 50-60 deg.c for at least 12 hr.
In some of these embodiments, the colloidal gold nanoparticle solution preparation method comprises: heating ultrapure water to 250 ℃ to boiling while stirring in an intelligent magnetic stirrer, adding 1/100-1/20 volume of sodium citrate with the mass concentration of 1wt%, and then rapidly adding HAuCl with the mass concentration of 1wt% 4 After continuing to heat for 10min-30min and after the color stability appears as reddish wine, the stable colloidal gold nanoparticle solution can be considered to be generated, cooled to room temperature and stored in the environment of 4 ℃ for standby.
In some of these embodiments, the step of preparing the colloidal Jin Xinxing coronavirus antibody complex using a colloidal gold nanoparticle solution specifically comprises: the pH value of the colloidal gold nanoparticle solution is regulated to 7-9, a novel coronavirus antibody is added into the colloidal gold nanoparticle solution with the mass concentration of 0.01wt%, the final concentration of the novel coronavirus antibody is 5 ug/mL-50 ug/mL, and the two are mixed and then subjected to oscillation reaction for 5-30 min; adding BSA until the final mass concentration of the BSA is 0.2%, and continuing to oscillate for 5-30 min; and (3) centrifuging to remove the supernatant to obtain a colloid Jin Xinxing coronavirus antibody complex, and re-dissolving the precipitate by adopting a gold-labeled diluent for later use.
Example 1
The present embodiment provides a novel coronavirus swab sample rapid detection kit 10. The preparation method of the swab sample rapid detection kit 10 comprises the following steps:
(1) The preparation method of the test strip 200 comprises the following steps:
preparing a conjugate pad: preparing a colloidal gold nanoparticle solution; the preparation method of the colloidal gold nanoparticle solution comprises the following steps: heating ultrapure water to 250 ℃ to boiling while stirring in an intelligent magnetic stirrer, adding 1/100-1/20 volume of sodium citrate with the mass concentration of 1wt%, and then rapidly adding HAuCl with the mass concentration of 1wt% 4 After continuing to heat for 30min, and after the color stability appears as a reddish wine, it can be considered that a stable colloidal gold nanoparticle solution has been generated, and the colloidal gold nanoparticle solution is cooled to room temperature and stored in an environment of 4 ℃ for standby.
Preparing a colloid Jin Xinxing coronavirus antibody complex by adopting a colloid gold nanoparticle solution; specifically, a colloidal gold nanoparticle solution is taken to adjust the pH value to 7-9, a novel coronavirus antibody is added into the colloidal gold nanoparticle solution with the mass concentration of 0.01wt%, the final concentration of the novel coronavirus antibody is 10ug/mL, and the two are mixed and then subjected to oscillation reaction for 30min; adding BSA until the final mass concentration of BSA is 0.2%, and continuing to oscillate for 30min; and (3) centrifuging to remove the supernatant to obtain a colloid Jin Xinxing coronavirus antibody complex, and re-dissolving the precipitate by adopting a gold-labeled diluent for later use.
The colloidal Jin Xinxing coronavirus antibody complex and the colloidal gold quality control antibody complex are diluted by a gold-labeled diluent to prepare a conjugate treatment solution, wherein the volume ratio of the colloidal Jin Xinxing coronavirus antibody complex in the conjugate treatment solution is 10%, the volume ratio of the colloidal gold quality control antibody complex is 2%, and the conjugate treatment solution is coated on a conjugate pad.
Preparation of nitrocellulose film: the novel coronavirus antigen capture antibody and the quality control capture antibody are respectively diluted to 1mg/mL and 1mg/mL by coating liquid, the coating liquid is dispensed on the corresponding position of the nitrocellulose membrane by a film drawing machine, and the nitrocellulose membrane is obtained by drying and fixing for 12 hours at 60 ℃, wherein the coating liquid comprises PBS with the value of 0.01mol/L, PH of 6.5-7.5 and 5% trehalose solution.
Preparing a sample pad: the treatment liquid is smeared on a sample pad, so that the liquid absorption amount is constant at 30+/-3 mL, and the sample pad is dried and fixed for 24 hours at 60 ℃ or naturally dried at room temperature for standby.
The sample pad, conjugate pad, nitrocellulose membrane and absorbent paper were sequentially adhered to a PVC base plate, and cut into 3 mm/strip of the novel coronavirus test strip 200 by a cutter under appropriate conditions of constant temperature and constant humidity.
(2) A connecting component 103 such as double faced adhesive tape is placed in the corresponding area of the second casing 102 of the first casing 101, a novel coronavirus test strip 200 is placed in the corresponding position of the second casing 102, and a swab component 300 is placed at the detection station 1012 of the first casing 101; the first shell 101 and the second shell 102 are correspondingly attached, wherein a detection line and a comparison line of the test strip 200 are positioned at the detection window 1011, and the first shell 101 and the second shell 102 are pressed together to form the complete rapid detection kit 10 for the novel coronavirus swab sample with the double-layer structure.
The sample swab is collected, the swab head of the sample swab is pressed in the first positioning groove 3011, the swab processing liquid is added above the swab head, then the swab rod is rapidly rotated, uniformly mixed and run, after standing for a set time, the detection result is read by the detection window 1011.
Example 2
The present embodiment provides a rapid detection kit 10 for a swab sample of influenza a/b virus. The preparation method of the swab sample rapid detection kit 10 comprises the following steps:
(1) The preparation method of the test strip 200 comprises the following steps:
preparing a conjugate pad: preparing a colloidal gold nanoparticle solution; the preparation method of the colloidal gold nanoparticle solution comprises the following steps: heating ultrapure water to 250 ℃ to boiling while stirring in an intelligent magnetic stirrer, adding 1/100-1/20 volume of sodium citrate with the mass concentration of 1wt%, and then rapidly adding HAuCl with the mass concentration of 1wt% 4 After continuing to heat for 30min, and after the color stability appears as a reddish wine, it can be considered that a stable colloidal gold nanoparticle solution has been generated, and the colloidal gold nanoparticle solution is cooled to room temperature and stored in an environment of 4 ℃ for standby.
Preparing a novel colloidal gold influenza A virus antibody compound, specifically, taking a colloidal gold nanoparticle solution, adjusting the pH value to 7-9, adding an influenza A antibody into the colloidal gold nanoparticle solution with the mass concentration of 0.01wt%, mixing the colloidal gold nanoparticle solution and the influenza A antibody, and then carrying out oscillation reaction for 30min, wherein the final concentration of the influenza A antibody is 10 ug/mL; adding BSA with the mass concentration of 20% to the final mass concentration of 0.2% of BSA, and continuing to oscillate for 30min; centrifuging to remove supernatant to obtain colloidal gold influenza A antibody complex, and redissolving the precipitate with gold-labeled diluent;
preparing a colloid Jin Yixing influenza virus antibody complex, specifically, taking a colloid gold nanoparticle solution, adjusting the pH value to 7-9, adding an influenza B virus antibody into the colloid gold nanoparticle solution with the mass concentration of 0.01wt%, mixing the two solutions, and then carrying out oscillation reaction for 30min; adding BSA with the mass concentration of 20% to the final mass concentration of 0.2% of BSA, and continuing to oscillate for 30min; and (3) centrifuging to remove the supernatant to obtain a colloid Jin Yixing influenza virus antibody complex, and re-dissolving the precipitate by adopting a gold-labeled diluent for later use.
The colloidal gold-labeled influenza A virus antibody complex, the colloidal Jin Yixing influenza virus antibody complex and the colloidal gold-labeled quality control antibody complex are diluted by a gold-labeled diluent to prepare a conjugate treatment solution, wherein the volume ratio of the colloidal gold-labeled influenza A virus antibody complex, the colloidal Jin Yixing influenza virus antibody complex and the colloidal gold-labeled quality control antibody complex is 10%, 10% and 2% in sequence, and the diluted conjugate treatment solution is coated on a conjugate pad, usually in a machine spraying, manual coating and other modes.
Preparation of nitrocellulose film: sequentially diluting an influenza A virus antigen capture antibody, an influenza B virus antigen capture antibody and a quality control capture antibody to 1.5mg/mL, 1.5mg/mL and 1.0mg/mL by using a coating liquid, spot-coating the coating liquid on the corresponding position of a nitrocellulose membrane by using a membrane dividing machine, and drying and fixing the coating liquid at 60 ℃ for 12 hours to obtain the nitrocellulose membrane, wherein the coating liquid comprises 0.5mol/L PBS with pH of 6.5-7.5 and 5% trehalose solution;
preparing a sample pad: smearing the treatment liquid on a sample pad to ensure that the liquid absorption capacity is constant at 30+/-3 mL, and drying and fixing for 24 hours at 60 ℃ for later use;
the sample pad, conjugate pad, nitrocellulose membrane and absorbent paper were sequentially adhered to a PVC base plate, and cut into 3 mm/strip influenza a/b virus test strips 200 by a cutter under appropriate conditions of constant temperature and constant humidity.
(2) A connecting component 103 such as double faced adhesive tape is placed in the corresponding area of the second casing 102 of the first casing 101, an influenza a/b virus test strip 200 is placed in the corresponding position of the second casing 102, and a swab component 300 is placed at the detection station 1012 of the first casing 101; the first casing 101 and the second casing 102 are correspondingly attached, wherein a detection line and a comparison line of the test strip 200 are positioned at the detection window 1011, and the first casing 101 and the second casing 102 are pressed together to form the complete rapid detection kit 10 for the swab sample of the influenza A/B virus with the double-layer structure.
The sample swab is collected, the swab head of the sample swab is pressed in the first positioning groove 3011, the swab processing liquid is added above the swab head, then the swab rod is rapidly rotated, uniformly mixed and run, after standing for a set time, the detection result is read by the detection window 1011.
Example 3
The rapid detection kit 10 for the novel coronavirus swab sample prepared in example 1 was used for detecting the nasal swab sample 400 of 50 volunteer patients with novel coronavirus symptoms randomly selected, men and women and ages were random, no drug treatment intervention was performed, no other basic diseases were caused, and the detection of the novel coronaantigen nucleic acid reagent, the detection of the traditional novel coronaantigen rapid reagent and the detection method using the novel coronaantigen detection card of the application were performed by the prior art, and the detection time and the detection result data were recorded as shown in table 1.
TABLE 1
The total detection time of the above nucleic acid detection, conventional rapid detection card, and swab sample rapid detection kit 10 of the novel coronavirus of the present application was collated to obtain the following table 2.
TABLE 2
Analysis of results: as can be seen from the data in tables 1 and 2, the accuracy of the rapid detection kit 10 for detecting the novel coronavirus swab sample in the embodiment 1 of the present application for detecting the novel coronavirus swab is basically consistent with that of the conventional nucleic acid detection technology, but the detection time is far longer than the detection efficiency of nucleic acid, the conventional reagent card has 4 cases of positive sample missing detection, and the sensitivity of the rapid detection kit 10 for detecting the novel coronavirus swab sample in the embodiment of the present application for detecting the novel coronavirus swab sample is better than that of the conventional reagent card detection technology, and the detection time is also better than that of the conventional reagent card detection efficiency.
Example 4
The rapid detection kit 10 for influenza a/b virus swab samples of example 2 was used to detect randomly selected nasal swab samples 400 of 50 volunteer patients with influenza symptoms, male and female and age were random, no drug treatment intervention was performed, no other underlying disease was caused, and the detection time and detection result data were recorded as shown in table 3.
TABLE 3 Table 3
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The total detection time of the above-described reagent card detection and the detection card of the present application was collated to obtain the following table 4.
TABLE 4 Table 4
As can be seen from the data in tables 3 and 4, the rapid detection kit 10 for the swab sample of the influenza A/B virus is used for detecting the antigen of the influenza A/B virus of the nose swab, and the accuracy is basically consistent with that of the traditional nucleic acid detection technology, but the detection time is far longer than that of the nucleic acid detection technology, 2 cases of positive samples are missed to be detected in the traditional reagent card A, 1 case of positive samples are missed to be detected in the B flow, and the detection card and the detection method are used for detecting the antigen of the nose swab, so that the sensitivity is better than that of the traditional reagent card detection technology, and the detection time is also better than that of the traditional reagent card detection efficiency.
In summary, according to the rapid detection kit 10 for the swab sample, the test strip 200 and the card are combined and assembled to form the reagent with the detection function, and the swab device is arranged at the sample adding end, so that the sample can be directly added for detection after sampling by the swab, the operation steps are shortened, the efficiency is improved, and the detection card for the swab sample 400 is prepared, so that the integration of direct sample adding and detection of the swab is realized, the operation is friendly, the sample is not easy to pollute, the detection efficiency is high, and the detection accuracy is high.
In the foregoing embodiments, the descriptions of the embodiments are emphasized, and for parts of one embodiment that are not described in detail, reference may be made to related descriptions of other embodiments.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The foregoing examples illustrate only a few embodiments of the application and are described in detail herein without thereby limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of protection of the present application is to be determined by the appended claims.
Claims (16)
1. The utility model provides a quick detection kit of swab sample, its characterized in that, includes kit main part, test paper strip and swab subassembly, the test paper strip with the swab subassembly all install in the kit main part, the swab subassembly has detection channel, detection channel's one end extends to the loading end of test paper strip, the loading end of test paper strip is located at least part in the detection channel, detection channel's the other end is towards the outside of kit main part for the supply swab sample to insert to contact the loading end of test paper strip.
2. The rapid detection kit for swab samples as claimed in claim 1, wherein a detection window is arranged on the kit main body, and the detection window is opposite to a detection line and a control line on the test strip.
3. The rapid test kit for a swab sample according to claim 1, wherein the kit body comprises a first housing and a second housing, the first housing is connected with the second housing and encloses a detection cavity, a detection station communicated with the detection cavity is arranged on the first housing or the second housing, the test strip and the swab assembly are both installed in the detection cavity, and the swab assembly is located at the detection station.
4. The rapid swab sample detection kit as recited in claim 3, wherein said first housing is provided with said detection station facing an interior surface of said detection chamber, said detection station being configured for mounting said swab assembly, said second housing being provided with a mounting station configured for mounting said test strip.
5. The rapid swab sample detection kit of claim 3, wherein the first housing is removably coupled to the second housing.
6. The rapid swab sample detection kit of claim 3, wherein the kit body further comprises a connection assembly, wherein at least one of the connection assemblies is connected to the first housing and/or the second housing, and wherein the first housing and the second housing are connected by the connection assembly.
7. The rapid test swab sample kit of claim 6, wherein the test strip is mounted to the second housing via the connection assembly, and wherein the swab assembly is mounted to the first housing via the connection assembly.
8. The rapid test kit of claim 6, wherein the connecting component is one or more of a double-sided tape, a magnetic alignment adsorption structure, and a rivet-fastening structure.
9. The rapid detection kit for a swab sample according to any one of claims 1 to 8, wherein a pressing mechanism for fixing a sample swab is arranged on the swab assembly, the pressing mechanism is provided with a first positioning groove for fixing a swab head of the sample swab and a second positioning groove for fixing a swab rod of the swab sample, the first positioning groove and the second positioning groove form the detection channel, and at least part of a loading end of the test strip is located in the first positioning groove.
10. The rapid swab sample detection kit of any one of claims 1-8, further comprising a swab sample, wherein the swab sample comprises a swab head and a swab stem coupled to the swab head, wherein the swab head is configured to sample.
11. The rapid swab sample detection kit of claim 10, wherein the swab sample comprises one or more of a nasal swab sample, a pharyngeal swab sample, a nasopharyngeal swab, a urethral swab, and a cervical swab sample.
12. The rapid detection kit for swab samples according to any one of claims 1 to 8 and 11, wherein the preparation material of the kit body is one or more selected from the group consisting of hard paper, polyethylene terephthalate, polymethyl methacrylate and polypropylene.
13. The rapid test kit for swab samples according to any one of claims 1 to 8 and 11, wherein the swab component is made of one or more materials selected from the group consisting of polyvinyl chloride, polyethylene terephthalate, polyethylene, polypropylene, polyvinylidene chloride, and polycarbonate.
14. A method for preparing a rapid test kit for a swab sample according to any one of claims 1 to 13, comprising the steps of:
(1) Preparing a test strip;
(2) The test strip is arranged at a preset position in the main body of the kit, the swab component is arranged at the preset position in the main body of the kit, one end of a detection channel of the swab component extends to the loading end of the test strip, so that the loading end of the test strip is at least partially positioned in the detection channel, and the other end of the detection channel faces the outside of the main body of the kit so as to be used for inserting a swab sample to be contacted with the loading end of the test strip.
15. The method for preparing the rapid detection kit for the swab samples according to claim 14, wherein the method for preparing the test strip comprises the following steps:
preparing a conjugate pad:
preparing a colloidal gold nanoparticle solution;
preparing a colloidal gold virus antibody complex by adopting the colloidal gold nanoparticle solution, diluting the colloidal gold virus antibody complex and the colloidal gold quality control antibody complex by using a gold-labeled diluent to prepare a conjugate treatment solution, and coating the conjugate treatment solution on a conjugate pad;
preparation of nitrocellulose film: respectively diluting a virus antigen capture antibody and a quality control capture antibody with coating liquid, then spot-coating the diluted virus antigen capture antibody and the quality control capture antibody on corresponding positions of a nitrocellulose membrane, and drying and fixing the diluted virus antigen capture antibody and the quality control capture antibody to obtain the nitrocellulose membrane;
preparing a sample pad: smearing the treatment liquid on a sample pad, and drying for later use;
and sequentially adhering the sample pad, the conjugate pad, the nitrocellulose membrane and the absorbent paper on a bottom plate, and cutting into test strips with preset sizes.
16. The method for preparing a rapid detection kit for a swab sample according to claim 15, wherein the volume ratio of the colloidal gold virus antibody complex in the conjugate treatment solution is 10% -20%, and the volume ratio of the colloidal gold quality control antibody complex is 2% -5%;
and/or the virus antigen capture antibody and the quality control capture antibody are respectively diluted to 0.5-2 mg/mL and 0.5-2 mg/mL by the coating liquid.
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