WO2023147713A1

WO2023147713A1 - Combined detection structure for dual sampling and integrated detection and use thereof

Info

Publication number: WO2023147713A1
Application number: PCT/CN2022/087799
Authority: WO
Inventors: 刘默文; 刘杰
Original assignee: 嘉兴康源科泰科技发展有限公司
Priority date: 2022-02-07
Filing date: 2022-04-20
Publication date: 2023-08-10
Also published as: CN114441755A

Abstract

Disclosed is a combined detection structure for dual sampling and integrated detection. The combined detection structure is a combined set of a saliva sampling structure (2), a nasal-cavity sampling structure (1) and a chromatographic assay structure (3), wherein the nasal-cavity sampling structure (1) is not only a collection structure for collecting a substance to be detected from the nasal cavity, but also a detection and sampling structure for detecting the substances to be detected in the saliva and the nasal cavity, the saliva sampling structure (2) is a collection structure for collecting the substance to be detected in saliva, and the substance collected from saliva is a solvent liquid phase of the substance to be detected in the nasal cavity and a mobile liquid phase for detecting the substance to be detected, and multi-functional designing that involves an inventive step is performed on the sampling structure. The combined detection structure can be used in the development of various rapid immunodetection products such as colloidal gold, fluorescent immunoassay and latex-microsphere immunoassay, which improves the detection efficiency, convenience and accuracy of immunodetection products, and has important clinical significance.

Description

A combined detection structure of double sample sampling and integrated detection and its application

technical field

The invention relates to the technical field of medical devices, in particular to a combined detection structure for double-sample sampling and integrated detection and its application.

Background technique

Immunological detection technology is an experimental method designed to determine antigens, antibodies, immune cells and chemical components based on the principle of applied immunology. , pharmaceutical analysis, food and industrial analysis samples. Commonly used detection techniques include immunoturbidity technology, solid-phase enzyme immunoassay technology, chemiluminescence detection technology, immunofluorescent labeling technology, quantum dot immunoassay technology, colloidal gold immunoassay technology, dot immunoassay technology, etc. High sensitivity, rapidity, convenience, miniaturization, full quantification, and automation are the current development trends of clinical immunoassay technology. Point-of-care testing (POCT) is currently the fastest-growing branch, and chromatography immunoassay is the most commonly used detection method, and colloidal gold, fluorescent lateral flow chromatography immunoassay and latex microsphere immunoassay technology are the most widely used , but the methods used are generally single-sample sampling and detection methods. As we all know, nasal swab or anterior nasal swab is the main sampling method for most respiratory infectious diseases at present. The extract is used for pretreatment of the sample. Saliva is also one of the test samples, but its viral load is lower than that of nasal swabs, which affects the positive detection rate of cases. Therefore, the development of dual-sample sampling technology for saliva and nasal cavity can not only superimpose the viral loads of the two collected samples, but also make saliva an effective solvent by optimizing the detection system, thereby realizing the realization that no extraction solution is required. The rapid detection realizes the accuracy, convenience, rapidity and simple operation of the detection, which will help the popularization and use of clinical detection products, improve the quality of diagnosis, treatment and prevention, and has important application value.

Contents of the invention

The purpose of the present invention is to provide a combined detection structure and application of double-sample sampling and integrated detection. Compared with the prior art, it has the advantages of high detection sensitivity, convenience, speed, and pollution prevention, and improves the detection quality.

In view of the above purpose, the present invention provides a combined detection structure of double-sample sampling and integrated detection, which has the following characteristics:

1) The combined detection structure is a combination suit of a saliva sampling structure, a nasal cavity sampling structure and a chromatography detection structure;

2) The saliva sampling structure is composed of a saliva collection structure and a saliva collection tube-like structure, which are detachably connected;

3) The nasal cavity sampling structure is composed of a sampling head and a sampling handle, and the sampling handle is provided with a sample loading connection structure that fixes the sampling head and forms a detachable connection with the chromatography detection structure;

4) The chromatographic detection structure includes a detection shell, a detection reagent strip located in the detection shell, and a detection connection structure located at the corresponding part of the starting end of the detection reagent strip.

In the above combined detection structure, the combined detection structure also has the following characteristics:

1) The combined suit formed by the saliva sampling structure, the nasal cavity sampling structure and the chromatography detection structure is a detachable structure;

2) The saliva sampling structure consists of a funnel-shaped saliva collection structure with a large top and a small bottom and a test tube-shaped saliva collection tube-like structure, with a detachable split structure between the two;

3) The nasal cavity sampling structure is a plug-in structure in which the sampling head is directly connected to the sampling handle and extends to the outside of the sampling handle;

4) The detection reagent strip placed inside the detection housing of the chromatographic detection structure is directly connected to the detection connection structure, and the detection connection structure includes the insertion inlet of the sampling head, and can form a removable connection with the sampling head. The inner connection structure of the detachable connection sample collection channel and the outer connection structure that can form a detachable connection with the sample loading connection structure;

5) The detection reagent strip includes a sample pad, a marker binding pad, a nitrocellulose membrane pad and an absorbent pad;

6) The internal connection structure of the detection connection structure contains a cavity structure that can form a liquid phase communication with the detection reagent strip, and its adjacent part is provided with a detection reagent strip placement layer, and the detection reagent strip is placed, wherein the The sample pad communicates with the cavity structure through a liquid flow opening, and the internal connection structure is composed of the insertion inlet of the sampling head, the sample collection channel and the liquid flow opening of the cavity structure and the detection reagent. The strips form a liquid communication link.

In the above-mentioned combined detection structure, the use state structure of the combined detection structure includes a detection sampling use structure and a detection reaction use structure, wherein the detection sampling use structure is to put the nasal cavity sampling structure with the nasal cavity sample collection into the collection Stir and mix the saliva sample in the saliva sampling structure to form an intermediate structure in which a double-sample mixture of nasal cavity sample and saliva sample is soaked and adsorbed on the nasal cavity sampling structure; The sampling head of the nasal cavity sampling structure for the nasal cavity and saliva samples to be tested is inserted into the detection connection structure, and then starts the use state intermediate structure of the detection reaction.

In the above combined detection structure, the detection shell is an up and down press-fit structure, including a top cover and a base, the proximal end of which is the detection connection structure, the base is provided with a bracket for placing a detection reagent strip, and the top cover The inner cavity of the inner cavity is provided with a sample collection channel for squeezing and collecting the sample to be tested. The detection reagent strip is arranged on the bracket, and is arranged in the direction of the sample pad, the marker binding pad, the nitrocellulose membrane pad and the water-absorbing pad. Arranged far and far in sequence, a liquid flow opening is arranged at the proximal end of the sample collection channel, which communicates with the sample pad of the detection reagent strip, and the upper cover is provided with an observation window, which is connected to the nitric acid of the detection reagent strip. The positions of the cellulose membrane pads are corresponding, and the proximal end of the press-fit structure is provided with a male-female insertion-type connecting structure. The detection structure end is a male-convex shape, and the sampling structure end is a female concave shape, or the detection structure end is a female concave shape, and the sampling structure end is a male-convex shape.

In the above combined detection structure, the detection shell is a sleeve-shaped structure, including an inner core and an outer sleeve, the outer surface of the inner core is provided with a bracket for placing the detection reagent strip, and the inner cavity of the inner core is provided with a In the sample collection channel of the sample to be tested collected by squeezing the sampling head, the detection reagent strip is arranged on the bracket, in the direction of the sample pad, marker binding pad, nitrocellulose membrane pad and water-absorbing pad Arranged in order from near to far, a liquid flow opening is provided at the proximal end of the sample collection channel and the bracket, which communicates with the sample pad of the detection reagent strip, and the outer sleeve is provided with an observation window, which is connected to the The position of the nitrocellulose membrane pad of the detection reagent strip is corresponding, and the proximal end of the sleeve-shaped structure is provided with the male-female insertion-type detection connection structure. The detection structure end is a male-convex shape, and the sampling structure end is a female concave shape, or the detection structure end is a female concave shape, and the sampling structure end is a male-convex shape.

In the above combined detection structure, the connecting part of the plug-in structure of the sampling head and the sampling handle is provided with a blocking structure to prevent liquid phase backflow, preferably a piston-type dynamic sealing washer structure made of elastic material. A piston-type dynamic sealing gasket structure made of elastic materials such as rubber gaskets and silicone gaskets is preferred.

In the above combined detection structure, the sampling head is inserted into the sample collection channel through the detection connection structure, and is squeezed at the far end, and the proximal end of the outer sleeve structure has a matching anti-sampling structure. The stacked plugging structure for liquid phase reflux is preferably a piston-type dynamic sealing gasket structure made of elastic material. A piston-type dynamic sealing gasket structure made of elastic materials such as rubber gaskets and silicone gaskets is preferred.

In the above combined detection structure, the saliva collection structure is selected from a funnel structure, a folding opening and closing structure or a telescopic opening and closing structure.

In the above combined detection structure, the detection reagent strip is selected from at least one of colloidal gold immunoassay, fluorescent immunoassay and latex microsphere immunoassay.

In the above combined detection structure, the operation and use of the combined detection structure includes the following steps:

1) Take the saliva sampling structure, spit the saliva into the saliva collection funnel-shaped structure, and collect the saliva sample through the saliva collection tube-like structure;

2) Take the nasal cavity sampling structure, insert the sampling head into the nasal cavity, and complete the sampling;

3) Put the sampling head of the nasal cavity sampling structure into the saliva collection tube-like structure to collect the saliva sample, stir repeatedly and soak to absorb the saliva sample;

3) Insert the sampling head containing the double samples of saliva and nasal cavity collection into the sample collection channel through the insertion inlet of the detection connection structure;

4) The liquid phase contained in the sampling head flows into the sample pad of the detection reagent strip through the liquid flow opening of the cavity structure, and then flows through the marker binding pad, nitrocellulose membrane pad, and water-absorbing pad to complete the detection.

5) Read the detection result from the observation window.

The saliva sampling structure of the combined detection structure collects saliva samples, the nasal cavity sampling structure collects nasal cavity samples, directly puts the sampling head with nasal cavity samples into the saliva collection sample, stirs and elutes, soaks and absorbs into the saliva sample. On the sampling head of the nasal cavity sampling structure, the sampling head soaked and absorbed with the nasal cavity and saliva sample mixture is directly loaded into the chromatographic detection structure through the detection connection structure to complete the sample loading and detection process.

The sampling head is made of water-absorbent and water-insoluble materials, including natural and modified polymer superabsorbent resins and artificially synthesized water-absorbent resins, including starch series, cellulose series, other natural product series, polyethylene salt series, polyvinyl alcohol series and polyoxyethylene series, etc.

The combined detection structure contains a detection aid solution, and the detection aid solution is a non-denaturing agent water quality buffer saline solution.

The detection reagent strip is a sample pad, a marker binding pad, a filter membrane pad, a nitrocellulose membrane pad and a water-absorbing pad that are sequentially pasted on the PVC film, and the detection reagent strip is placed in the detection shell.

The present invention has the following advantages due to the adoption of the above technical scheme:

1. The present invention is a rapid detection structure that adopts double sample sampling of saliva and nasal cavity, and collects double samples of saliva and nasal cavity at the same time during detection, which can significantly improve the positive detection rate of cases and improve the detection sensitivity of diseases, which is not like the existing There is a market that can only make a fuss about improving the sensitivity of detection reagents. As we all know, nasal swab or anterior nasal swab is the main sampling method for most respiratory infectious diseases at present. The extract is used for pretreatment of the sample. Saliva is also one of the test samples, but its viral load is lower than that of nasal swabs, which affects the positive detection rate of cases. The present invention adopts double-sample sampling of saliva and nasal cavity, which can not only superimpose the viral load in the two collected samples, but also make saliva an effective solvent for use by optimizing the detection system, thereby achieving the realization that no The rapid detection of the extract realizes the superior accuracy, convenience, rapidity and simple operation of the detection reagent.

2. The nasal cavity sampling structure of the present invention is not only the collection structure of the nasal cavity to be detected, but also the detection and sample addition structure for detecting saliva and nasal cavity to be detected. The saliva sampling structure is not only the collection structure of the saliva to be detected, but also It is also the solvent liquid phase of the substance to be tested in the nasal cavity and the mobile liquid phase for the detection of the substance to be tested, and a creative multifunctional design has been carried out for the sampling structure.

3. The saliva sampling structure of the present invention is a funnel-shaped structure with a collection tube, so that the saliva can be spit out through the funnel and directly enter the collection tube, which is easy to operate and avoids pollution.

4. The sampling structure of the nasal cavity sampling of the present invention includes a sampling handle and a sampling head, wherein the sampling head has dual functions of sampling and detection and loading, and has various advantages such as convenient collection, sample transfer and sample loading safety.

5. The detection connection structure adopted in the present invention is a structure in which the sample collection channel is adjacent to the reagent detection channel, and there is a liquid-phase flow traffic structure at the beginning of the reagent strip, which not only ensures the accuracy of sampling, but also ensures The structure and detection function of the detection reagent strip are not affected during the sampling process.

6. The present invention adopts the detection test strip as the detection component, and is applicable to multiple detection technologies with lateral flow as the main technology, such as colloidal gold, fluorescent immunoassay, latex microsphere immunoassay, etc., which expands the scope of application.

7. The present invention is equipped with an auxiliary detection liquid phase, which can be used for samples with insufficient liquid phase, and improves the practicality of detection.

8. The invention has simple operation steps and is easy to realize home use or self-testing. It is not only convenient to use and reduces the waste of raw materials, but also significantly improves work efficiency, and can be applied to many fields of professional and amateur testing.

Description of drawings

Fig. 1 is the overall structure schematic diagram of the present invention;

Fig. 2 is a schematic structural diagram of the connection between components in the combined use state of the present invention;

Fig. 3 is the structural representation of saliva sampling of the present invention;

Fig. 4 is a schematic diagram of the foldable saliva sampling structure of the present invention;

Fig. 5 is a schematic diagram of the scalable saliva sampling structure of the present invention;

Fig. 6 is a schematic diagram of the up and down press-fit detachable structure of the present invention;

Fig. 7 is a structural schematic diagram of the upper and lower press-fit longitudinal sections of the present invention;

Fig. 8 is a schematic diagram of the upper and lower press-fit chromatography detection structure of the present invention;

Fig. 9 is a schematic diagram of a sleeve-shaped structure of the present invention;

Fig. 10 is a schematic structural view of a sleeve-shaped longitudinal section of the present invention;

Fig. 11 is a schematic diagram of the sleeve-shaped detachable structure of the present invention.

The markings in the figure are as follows:

Nasal cavity sampling structure 1; saliva sampling structure 2; chromatography detection structure 3; saliva collection structure 4; saliva collection tube 5; sampling handle 6; sampling connection structure 7; sampling fixed structure 8; sampling head 9; Reagent strip 11; observation window 12; detection shell 13; detection connection sampling head insertion port 14; complex sampling use structure 15; detection reaction use structure 16, saliva collection tube cover 17, foldable saliva collection structure use state 18, foldable Preservation state of sexual saliva collection structure 19, use state of scalable saliva collection structure 20, storage state of scalable saliva collection structure 21, upper and lower pressing type sampling structure handle 22, upper and lower pressing type sampling fixed structure 23, upper and lower pressing type detection connection structure 24. Up and down press-fit detection cavity structure 25, up and down press-fit sampling connection structure 26, up and down press-fit detection internal connection structure 27, up and down press-fit detection liquid flow opening 28, up and down press-fit detection sample collection channel 29, up and down press-fit type Detection reagent strip bracket 30, upper and lower press-fit chromatography detection structure upper cover 31, detection reagent strip sample pad 32, detection reagent strip marker binding pad 33, detection reagent strip nitrocellulose membrane pad 34, detection reagent strip absorbent pad 35 , upper and lower press-fit chromatography detection structure base 36, sleeve-shaped detection structure handle 37, sleeve-shaped detection sampling connection structure 38, sleeve-shaped sampling fixed structure 39, sleeve-shaped detection sampling blocking structure 40, sleeve-shaped detection Detection connection structure 41, sleeve-shaped detection cavity structure 42, sleeve-shaped detection structure inner core 43, sleeve-shaped detection structure outer sleeve 44, sleeve-shaped detection reagent strip bracket 45, sleeve-shaped detection sample collection The channel 46 and the liquid flow opening 47 of the sleeve-shaped detection structure.

Detailed ways

In order to further illustrate the technical means and effects adopted by the present invention to achieve the intended purpose, the following embodiments will further illustrate the present invention in conjunction with the accompanying drawings, but the present invention is not limited to the following description.

As shown in Figure 1 and Figure 2, the overall structure of the present invention includes a nasal cavity sampling structure 1, a saliva sampling structure 2 and a chromatography detection structure 3, wherein the saliva sampling structure 2 includes a saliva collection structure 4 and a saliva collection tube 5, and the saliva collection structure 4 It is a funnel-shaped structure, and the small mouth of the funnel of the saliva collection structure 4 forms a detachable connection with the saliva collection tube 5; the nasal cavity sampling structure 1 includes a sampling handle 6, a sampling connection structure 7, a sampling fixing structure 8 and a sampling head 9; chromatography detection The structure 3 includes a detection shell 13 , a detection connection structure 10 , a sampling head insertion port 14 , a detection reagent strip 11 and an observation window 12 . The saliva is collected into the saliva collection tube 5 through the saliva collection structure 4, and then the nasal cavity sample is collected with the sampling head 9, and the sampling head 9 with the nasal cavity sample collected is directly put into the saliva collection tube 5 with the saliva sample collected, and stirred and eluted The mixture of saliva and nasal cavity samples is soaked and absorbed on the sampling head 9, and the sampling head 9 is immersed in the saliva collection tube 5 to form a composite sampling structure 15. Then take out the sampling head 9 soaked and absorbed the mixture of nasal cavity and saliva samples, directly insert it into the detection connection structure 10 through the sampling head insertion port 14, and further add the mixed sample to the detection reagent strip 11 in the chromatography detection structure 3 , start the detection reaction and complete the detection. At this time, the sampling head 9 is inserted into the detection connection structure 10 through the sampling head insertion port 14 to form a stable connection detection reaction use structure 16 . Then check the detection result through the observation window 11, and then realize the integrated operation of double-sample sampling and detection. The detection structure of the present invention is an integrated structure, and the double-sample sampling and detection are completed on the same structure.

As shown in Fig. 3, Fig. 4, and Fig. 5, in the saliva collection structure 4 and the saliva collection tube 5 of the saliva sampling structure 2 of the present invention, the saliva collection tube 5 is a tubular structure, which is also equipped with saliva in addition to the saliva collection tube 5. The collection tube cover 17; the saliva collection structure 4 can be a common funnel, a foldable opening and closing funnel and a telescopic opening and closing funnel. The shape of the ordinary funnel is relatively fixed and remains unchanged during use. The states of the foldable opening and closing funnel include the foldable saliva collection structure storage state 19 and the foldable saliva collection structure use state 18, which are respectively used for transportation, storage and sampling. The state of the retractable opening and closing funnel includes the storage state 21 of the retractable saliva collection structure and the use state 20 of the retractable saliva collection structure, which are respectively used for transportation, storage and sampling.

As shown in Fig. 6, Fig. 7 and Fig. 8, the detection shell 13 in the combined detection structure of the present invention is an up and down press-fit structure, including an upper and lower press-fit tomographic detection structure upper cover 31, and an up and down press-fit tomographic detection structure base 36. The base 36 of the upper and lower press-fit chromatography detection structure is provided with an upper and lower press-fit detection reagent strip bracket 30 for placing the detection reagent strip 11, and a sample pad 32 and a marker binding pad 33 are sequentially placed in the upper and lower press-fit detection reagent strip bracket 30 , nitrocellulose membrane pad 34 and absorbent pad 35. The inner cavity of the upper and lower pressure-fit chromatography detection structure upper cover 31 is provided with an upper and lower pressure-fit detection sample collection channel 29 for squeezing and collecting samples to be tested, and the upper and lower pressure-fit detection sample collection channels 29 are located close to the insertion port 14 of the detection connection sampling head The upper and lower press-fit detection cavity structure 25 is provided with the upper and lower press-fit detection liquid flow openings 28 . The observation window 12 corresponds to the position of the nitrocellulose membrane pad 34 of the detection reagent strip 11 . Located at the proximal end of the up and down press-fit detection structure, a male-female plug-in type up and down press-fit detection connection structure 24 and an up and down press-fit sampling connection structure 26 are provided. The detection structure end is a male-convex shape, and the sampling structure end is a female concave shape, or the detection structure end is a female concave shape, and the sampling structure end is a male-convex shape.

As shown in FIG. 9 , FIG. 10 , and FIG. 11 , the detection shell 13 in the combined detection structure of the present invention is a sleeve-shaped structure, including a sleeve-shaped detection structure inner core 43 and a sleeve-shaped detection structure outer sleeve 44 . The outer surface of the inner core 43 of the sleeve-shaped detection structure is provided with a sleeve-shaped detection reagent strip bracket 45 for placing the detection reagent strip 11. Sleeve-shaped test sample collection channel 46 for collected samples to be tested. There is a sleeve-shaped detection cavity structure 42 with a sleeve-shaped detection structure liquid flow opening 47 at the position of the sleeve-shaped detection sample collection channel 46 close to the insertion port 14 of the detection connection sampling head. The observation window 12 corresponds to the position of the nitrocellulose membrane pad of the detection reagent strip 11 . The proximal end of the sleeve-shaped detection structure is provided with a male-female insertion-type sleeve-shaped detection connection structure 41 .

In actual operation, when the rapid detection structure of the present invention is a colloidal gold immune detection structure, the detection reagent strip 11 is prepared by the colloidal gold method, and the colloidal gold of the sample pad and the colloidal gold marker are pasted on the PVC film successively. A combination pad, a nitrocellulose membrane pad and a water-absorbent pad coated with a non-labeled capture reagent; when the rapid detection structure of the present invention is a latex microsphere immunoassay structure, the detection reagent strip 11 is prepared by a latex microsphere immunoassay, and is placed on a PVC film A sample pad, a latex microsphere binding pad coated with a latex microsphere marker, a nitrocellulose membrane pad coated with a non-labeled capture reagent, and an absorbent pad are pasted on the surface in sequence. Use the saliva sampling structure 2, the nasal cavity sampling structure 1 and the chromatography detection structure 3 in combination. When in use, first use the saliva sampling structure 2 to collect a saliva sample, take the nasal cavity sampling structure 1 to collect a nasal cavity sample, then put the sampling head 9 of the nasal cavity sampling structure 1 into the saliva collection tube 5, stir and mix and absorb the saliva mixture to the sampling head 9 Then, the sampling head 9 is inserted into the sample collection channel through the sampling insertion port 14. At this time, the sampling head 9 is squeezed, and the liquid phase mixed sample on the sampling head 9 flows through the liquid flow opening of the cavity structure to the The sample pad of the detection reagent strip 11 passes through the marker binding pad, the nitrocellulose membrane pad and the water-absorbing pad to start the detection reaction, and then the detection result is read through the observation window 12 .

Experimental research of the present invention: the following experiments illustrate the detection method and its effects of the present invention, but are not limiting to the present invention. The experimental methods used in the following experiments are conventional methods unless otherwise specified. The materials and reagents used can be obtained from commercial sources unless otherwise specified.

Experiment 1: Rapid detection of new coronavirus antigen by immunocolloidal gold method:

1. Detection reagent strip preparation:

The detection reagent strip was prepared by the double-antibody sandwich method of conventional immune colloidal gold detection technology, and the detection kit was prepared by using the combined detection structure of the present invention to carry out the new coronavirus antigen detection experiment. The anti-new coronavirus S protein monoclonal antibody in ml is made of colloidal gold particles with a particle size of 50nm, coated on the glass cellulose membrane colloidal gold binding pad; the detection line T of the detection reagent strip has a paired capture antibody of 1.0mg/ml Anti-new coronavirus S protein monoclonal antibody, coated on nitrocellulose membrane pad; the capture antibody of the quality control line C of the detection reagent strip is 1.0mg/ml goat anti-mouse IgG polyclonal antibody, coated on nitrocellulose membrane The pad is used to capture the colloidal gold-labeled anti-new coronavirus S protein monoclonal antibody that has not been specifically captured. Paste absorbent pads and colloidal gold marker binding pads on both ends of the nitrocellulose membrane pad printing membrane, and paste sample pads on the side of the colloidal gold marker binding pad. Place the pasted test piece on the strip cutter and cut it into 3.5mm test reagent strips.

2. Preparation of the combined detection structure of the present invention:

Adopt Solidworks to design the upper cover, base, nasal cavity sampling structure and folded saliva sampling structure of the combined detection structure detection shell of the present invention, and use 3D printing samples, paste the sponge sampling head on the sampling head fixed structure, and use the preservation tube used in small experiments as Saliva collection tube, the prepared combined detection structure sample is used for experimental detection.

3. Experimental methods and results:

During the experiment, take the detection reagent strip and combined detection structure prepared above, assemble it into a detection buckle, put the assembled detection buckle into an aluminum foil sealed bag with a desiccant, seal it on a sealing machine, and affix a label. Prepare two solutions to be tested, 1) use the saliva of healthy people to prepare different concentrations of recombinant new coronavirus antigen S protein solutions; Recombinant SARS-CoV-2 antigen S protein solution. Two methods were used for detection. Method 1: Take 150ul solution 1), add it into a test tube, take a nasal cavity sampling structure, collect nasal cavity samples from healthy people, insert the sampling head into the solution 1), stir and absorb the solution 1), and then pass through Insert the insertion port into the detection buckle, let it stand for 20 minutes, check the observation window, and read the color development result on the detection reagent strip. Method 2: Take 100ul of saliva from a healthy person without S protein, add it to the test tube, take the nasal cavity sampling structure, collect a healthy human nasal cavity sample, and then take 50ul of the solution 2) Add it to the sampling head after nasal cavity sampling, insert the sampling head into In the saliva of the blank healthy person, stir and absorb the saliva solution of the blank healthy person, then insert it into the detection buckle through the insertion port, let it stand for 20 minutes, check the observation window, and read the color development result on the detection reagent strip. The quality control line C of the detection reagent strip has color development, and the detection line T has no color development, which is negative; the quality control line C has color development, and the detection line T also has color development, which is positive. When the concentration of antigen S protein solution is 1.0, 0.1, 0.05ng/ml, the detection results of the two methods are all positive, and when the concentration is 0.01ng/ml or below, the detection results are all negative.

Experiment 2: Rapid detection of new coronavirus antigen by latex microsphere immunochromatography:

1. Detection reagent strip preparation:

Using conventional latex microsphere immunochromatography technology double antibody sandwich method to prepare detection reagent strips, using the combination detection structure of the present invention to prepare detection kits for new coronavirus antigen detection experiments, wherein the latex microspheres are all biological 300nm red microspheres, detection reagents The latex microsphere label indicator antibody of the detection line T of the strip is 50ug/ml anti-new coronavirus S protein monoclonal antibody, which is coated on the glass cellulose membrane binding pad; the capture antibody of the detection line T of the detection reagent strip is 1.0mg/ml The paired anti-new coronavirus S protein monoclonal antibody in ml is coated on the nitrocellulose membrane pad; the capture antibody of the quality control line C of the detection reagent strip is 1.0 mg/ml goat anti-mouse IgG polyclonal antibody, coated on the On the nitrocellulose membrane pad, it is used to capture the latex microspheres labeled anti-new coronavirus S protein monoclonal antibody that has not been specifically captured. Paste absorbent pads and latex microsphere marker binding pads on both ends of the nitrocellulose membrane pad printing membrane, and paste sample pads on one side of the latex microsphere marker binding pad. Place the pasted test piece on the strip cutter and cut it into 3.5mm test reagent strips.

2. Preparation of the combined detection structure of the present invention:

Prepared with "Experiment 1".

3. Experimental methods and results:

During the experiment, take the detection reagent strip and combined detection structure prepared above, assemble it into a detection buckle, put the assembled detection buckle into an aluminum foil sealed bag with a desiccant, seal it on a sealing machine, and affix a label. As above, prepare two kinds of solutions to be tested, 1) use the saliva of healthy people to prepare different concentrations of recombinant new coronavirus antigen S protein solution; 2) use non-inactivated virus preservation solution (UTM) to prepare three times the concentration of the solution Concentration of recombinant new coronavirus antigen S protein solution. Two methods were used for detection. Method 1: Take 150ul solution 1), add it into a test tube, take a nasal cavity sampling structure, collect nasal cavity samples from healthy people, insert the sampling head into the solution 1), stir and absorb the solution 1), and then pass through Insert the insertion port into the detection buckle, let it stand for 20 minutes, check the observation window, and read the color development result on the detection reagent strip. Method 2: Take 100ul of saliva from a healthy person without S protein, add it to the test tube, take the nasal cavity sampling structure, collect a healthy human nasal cavity sample, and then take 50ul of the solution 2) Add it to the sampling head after nasal cavity sampling, insert the sampling head into In the saliva of the blank healthy person, stir and absorb the saliva solution of the blank healthy person, then insert it into the detection buckle through the insertion port, let it stand for 20 minutes, check the observation window, and read the color development result on the detection reagent strip. The quality control line C of the detection reagent strip has color development, and the detection line T has no color development, which is negative; the quality control line C has color development, and the detection line T also has color development, which is positive. When the concentration of antigen S protein solution is 1.0, 0.1, 0.05ng/ml, the test results of the two methods are all positive, and when the concentration is 0.01ng/ml or below, the test results are all negative.

Claims

A combined detection structure for double-sample sampling and integrated detection, characterized in that:

1) The combined detection structure is a combination suit of a saliva sampling structure, a nasal cavity sampling structure and a chromatography detection structure;

2) The saliva sampling structure is composed of a saliva collection structure and a saliva collection tube-like structure, which are detachably connected;

3) The nasal cavity sampling structure is composed of a sampling head and a sampling handle, and the sampling handle is provided with a sample loading connection structure that fixes the sampling head and forms a detachable connection with the chromatography detection structure;

4) The chromatographic detection structure includes a detection shell, a detection reagent strip located in the detection shell, and a detection connection structure located at the corresponding part of the starting end of the detection reagent strip.
Combined detection structure as claimed in claim 1, is characterized in that:

1) The combined suit formed by the saliva sampling structure, nasal cavity sampling structure and chromatography detection structure is a detachable structure;

2) The saliva sampling structure consists of a funnel-shaped saliva collection structure with a large top and a small bottom and a test tube-shaped saliva collection tube-like structure, which can be disassembled;

3) The nasal cavity sampling structure is directly fixedly connected between the sampling head and the sampling handle;

4) The detection reagent strip placed inside the detection shell of the chromatography detection structure is directly connected to the detection connection structure, and the detection connection structure includes the sampling head insertion port, which can form a detachable connection with the sampling head. The inner connection structure of the sample collection channel connected in the loading mode and the outer connection structure that can form a detachable connection with the sample loading connection structure;

5) The detection reagent strip includes a sample pad, a marker binding pad, a nitrocellulose membrane pad and an absorbent pad;

6) The internal connection structure of the detection connection structure contains a cavity structure that can form a liquid phase communication with the detection reagent strip, and a detection reagent strip is placed at the adjacent part, wherein the sample pad communicates with the cavity structure, so The insertion port of the sampling head is connected with the sample collection channel and the liquid phase communication between the cavity structure and the detection reagent strip.
The combined detection structure according to claim 1, wherein the use state structure of the combined detection structure includes a detection sampling use structure and a detection reaction use structure, wherein the detection sampling use structure is the nasal cavity sample that will be collected. The nasal cavity sampling structure is put into the saliva sampling structure with the saliva sample collected, stirred and mixed to form a double-sample mixture of the nasal cavity sample and the saliva sample, and the mixture is soaked in the use-state intermediate structure adsorbed on the nasal cavity sampling structure; the detection The reaction use structure is an intermediate structure in the use state that inserts the sampling head of the nasal cavity sampling structure soaked and adsorbed with nasal cavity and saliva samples to be tested into the detection connection structure, and then starts the detection reaction.
The combined detection structure according to claim 1, characterized in that, the detection shell is a press-fit structure up and down, including an upper cover and a base, the base is provided with a bracket for placing a detection reagent strip, and the inner cover of the upper cover is The cavity is provided with a sample collection channel for squeezing and collecting the sample to be tested. The sample pad, the marker binding pad, the nitrocellulose membrane pad and the water-absorbing pad are sequentially placed in the bracket, and a liquid is arranged at the proximal end of the sample collection channel. Open a hole so that the sample collection channel communicates with the sample pad of the detection reagent strip, the upper cover is provided with an observation window corresponding to the position of the nitrocellulose membrane pad of the detection reagent strip, the The proximal end of the press-fit structure is provided with a male-female insertion type connecting structure.
The combined detection structure according to claim 1, wherein the detection shell is a sleeve-shaped structure, including an inner core and an outer sleeve, and the outer surface of the inner core is provided with a bracket for placing a detection reagent strip, so The inner cavity of the inner core is provided with a sample collection channel for extruding the sample to be tested collected by the sampling head, and the sample pad, the marker binding pad, the nitrocellulose membrane pad and the water-absorbing pad are sequentially placed in the bracket, A liquid flow opening is arranged at the proximal end of the sample collection channel and the bracket, so that the sample collection channel communicates with the sample pad of the detection reagent strip, and the outer sleeve is provided with an observation window, which is connected to the detection reagent strip. The position of the nitrocellulose membrane pad of the reagent strip is corresponding, and the proximal end of the sleeve-like structure is provided with the male-female insertion-type detection connection structure.
The combined detection structure according to claim 1, characterized in that, the connecting part of the plug-in structure of the sampling head and the sampling handle is provided with a blocking structure to prevent liquid phase backflow, and the blocking structure is made of elastic material. A piston-type dynamic sealing gasket.
The combined detection structure according to claim 1 or 5, characterized in that, the sampling head is inserted into the sample collection channel through the detection connection structure, and is squeezed at the far end, and the proximal end of the outer sleeve structure has A nested plugging structure that matches the sampling structure to prevent liquid phase backflow, and the stacked plugging structure is a piston-type dynamic sealing gasket made of elastic material.
The combined detection structure according to claim 1, wherein the saliva collection structure is a funnel structure, a foldable opening structure or a telescopic opening structure.
The combined detection structure according to claim 1, wherein the detection reagent strip is at least one of colloidal gold immunoassay, fluorescent immunoassay and latex microsphere immunoassay.
The combined detection structure according to claim 1, wherein the operation and use of the combined detection structure comprises the steps of:

1) Take the saliva sampling structure, spit saliva into the saliva collection structure, and collect the saliva sample through the saliva collection tube-like structure;

2) Take the nasal cavity sampling structure, insert the sampling head into the nasal cavity, and complete the sampling;

3) Put the sampling head of the nasal cavity sampling structure into the saliva collection tube-like structure to collect the saliva sample, stir repeatedly and soak to absorb the saliva sample;

3) Insert the sampling head containing the double samples of saliva and nasal cavity into the sample collection channel;

4) The liquid phase contained in the sampling head flows into the sample pad of the detection reagent strip, and then flows through the marker binding pad, nitrocellulose membrane pad, and water-absorbing pad to complete the detection;

5) Read the test result.
The application of the combined detection structure as claimed in claim 1 in rapid detection product development.
Combined detection structure as claimed in claim 1, is characterized in that:

1) The combined detection structure is a combination suit of a saliva sampling structure, a nasal cavity sampling structure and a chromatography detection structure;

2) The saliva sampling structure is composed of a saliva collection structure and a saliva collection tube-like structure, which are detachably connected;

3) The nasal cavity sampling structure includes a sampling head and a sampling handle, the sampling handle is fixedly connected to the sampling head, and the sampling handle is detachable from the chromatography detection structure;

4) The chromatographic detection structure includes a detection housing, a detection reagent strip disposed in the detection housing, and a detection connection structure inserted into the corresponding part of the starting end of the detection reagent strip.