SUMMERY OF THE UTILITY MODEL
An object of the utility model is to provide a novel coronavirus and influenza virus detect reagent box has easy operation, weak point consuming time and efficient, detect that required sample size is few, detectivity is high, advantage such as with low costs.
In order to achieve the above object, the utility model provides a novel coronavirus and influenza virus detect reagent box, including the reagent box body that is equipped with the box chamber, along the length direction of the reagent box body sample pad, blood filtering pad, conjugate pad, nitrocellulose membrane and absorbent filter paper have arranged in proper order in the box chamber, the conjugate pad is coated with novel coronavirus monoclonal detection antibody and quality control thing of marker mark, the nitrocellulose membrane includes detection zone and quality control area that the interval set up, the detection zone is coated and is fixed with novel coronavirus monoclonal capture antibody of discerning single epitope, the quality control area is coated and is fixed with quality control capture antibody; and a sample adding port corresponding to the sample pad and a display window corresponding to the detection area and the quality control area are arranged on the box cover of the reagent box body.
Preferably, the combination pad is also coated with influenza A and/or B virus monoclonal detection antibodies, and the detection area is correspondingly coated and fixed with the influenza A and/or B virus monoclonal capture antibodies.
Preferably, the detection area is provided with coating lines arranged at intervals, and the novel coronavirus monoclonal capture antibody, the influenza a virus monoclonal capture antibody and/or the influenza b virus monoclonal capture antibody and the coating lines are in one-to-one correspondence and are respectively coated and fixed on the corresponding coating lines to form a novel coronavirus detection zone, an influenza a virus detection zone and/or an influenza b virus detection zone.
Preferably, the coating amount of the novel coronavirus monoclonal capture antibody and the quality control capture antibody is 0.01-5 mu g/cm.
Preferably, the quality control substance is set as a rabbit lgG antibody, and the quality control capture antibody is set as a goat anti-rabbit lgG antibody.
Preferably, the marker is any one of fluorescent microsphere, colloidal gold, magnetic particle or colored microsphere.
Preferably, the ends of the sample pad, the blood filter pad, the combination pad, the nitrocellulose membrane and the water-absorbing filter paper, which are arranged in sequence, are overlapped with each other.
Preferably, the label is loaded with a lanthanide or a chelate thereof.
Preferably, the lanthanide is samarium, europium or terbium.
Preferably, the sample pad, the blood filtration pad, the combination pad, the nitrocellulose membrane and the absorbent filter paper are all adhered to the bottom plate.
The utility model provides an among the technical scheme, novel coronavirus and influenza virus detect reagent box, including the reagent box body that is equipped with the box chamber, arrange in proper order along the length direction box intracavity of reagent box body and set up sample pad, blood filtering pad, combination pad, nitrocellulose membrane and absorbent filter paper, the combination pad is filled up and is wrapped up novel coronavirus monoclonal detection antibody and the quality control thing that has the marker mark, and nitrocellulose membrane includes detection zone and the quality control district that parallel and interval set up, and the detection zone is wrapped up and is fixed with novel coronavirus monoclonal capture antibody of discerning single epitope, and the quality control district is wrapped up and is fixed with the quality control capture antibody; the box cover of the reagent box body is provided with a sample adding port corresponding to the sample pad and a display window corresponding to the detection area and the quality control area.
When in use, a sample to be detected is dripped to the sample pad from the sample adding port, and then sample diluent is dripped; the sample moves from the sample pad to the absorbent filter paper along each attachment on the bottom plate of the kit in sequence, the virus antigen is respectively subjected to immunological binding with the detection antibody and the capture antibody and captured, the quality control substance is combined with the quality control capture antibody and captured, and the color of the marker visible to naked eyes can be displayed after sufficient capture. If the detection area and the quality control area both show the color carried by the marker, the sample is positive to the novel coronavirus; if only the quality control area displays the color carried by the marker, the sample is negative to the novel coronavirus; if the detection area and the quality control area do not display the color carried by the marker, the detection is invalid and needs to be detected again.
The utility model provides a novel coronavirus and influenza virus detect reagent box, can simplify the detection procedure of novel coronavirus, do not have the dependency to any laboratory glassware, environment or operating personnel, easy operation, convenience, detection cycle is short, high efficiency, easy to interpret; the kit needs a small amount of samples for detection, performs detection through antibody reaction, has high detection sensitivity, and has the advantages of low cost and the like.
The utility model provides an in the preferred scheme, still the parcel has influenza A and/or type B virus monoclonal antibody detection on the combination pad of detect reagent box, and the detection zone correspondence of nitrocellulose membrane is wrapped and is fixed with influenza A and/or type B virus monoclonal antibody capture. Then the utility model provides a novel coronavirus and influenza virus detect reagent box can high efficiency detect the sample to once can carry out clinical differentiation to three kinds of viruses, confirm the virus type that carries in the sample, carry out the diagnosis of once only distinguishing fast to three kinds of diseases that clinical symptom and complication have the similarity, simplify the testing process, improve and confirm diagnosis efficiency.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be described in detail below. It is to be understood that the embodiments described are only some embodiments of the invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.
The specific embodiment aims to provide a novel coronavirus and influenza virus detection kit, which has the advantages of simple operation, short time consumption, high efficiency, small sample amount required for detection, high detection sensitivity, low cost and the like.
Hereinafter, embodiments will be described in detail with reference to the accompanying drawings. The embodiments described below do not limit the scope of the invention described in the claims. Further, the entire contents of the configurations shown in the following embodiments are not limited to those necessary as a solution of the invention described in the claims.
Referring to fig. 1-6, in the present embodiment, a reagent kit for detecting coronavirus and influenza virus is provided, as shown in fig. 2, a reagent kit body 7 is composed of a bottom plate 6, a side wall and a cover plate, and is provided with a kit cavity. As shown in fig. 1, a sample pad 5, a blood filtration pad 4, a combination pad 3, a nitrocellulose membrane 2 and water-absorbing filter paper 1 are sequentially arranged in a chamber along the length direction of a reagent cartridge body 7, and all of these attachments can be adhered to a bottom plate 6 for fixation, and the bottom plate 6 can be made of a PVC plate. The gap between the cover and the bottom plate 6 promotes the liquid to flow rapidly on these attachments under the action of capillary. And between the sample pad 5, the blood filter pad 4, the combination pad 3, the nitrocellulose membrane 2 and the water-absorbing filter paper 1 which are arranged in sequence, the end parts of the two adjacent to each other can be mutually abutted or overlapped, for example, the end parts of the two adjacent to each other are pressed together and bonded, so as to strengthen the safety transition of the liquid flowing between the two adjacent to each other. The sample pad 5 and the conjugate pad 3 are both sample pads and conjugate pads used in the prior art colloidal gold method, and may be glass fiber membranes.
The binding pad 3 is coated with a novel coronavirus (2019-nCoV) monoclonal detection antibody and a quality control substance which are marked by a marker, wherein the marker can be a substance which can be combined with the antibody and has a color display, such as fluorescent microspheres, colloidal gold, magnetic particles or colored microspheres. The nitrocellulose membrane 2 comprises a detection area and a quality control area which are arranged in parallel at intervals along the direction from the sample pad 5 to the water absorption filter paper 1, as shown in figure 2, the detection area is close to the combination pad 3, the quality control area is close to the water absorption filter paper 1, the detection area is coated and fixed with a novel coronavirus (2019-nCoV) monoclonal capture antibody for identifying a single epitope, and the quality control area is coated and fixed with a quality control capture antibody. The quality control substance is a substance which can be specifically combined with the quality control capture antibody but can not be combined with the novel coronavirus monoclonal capture antibody, such as any protein meeting the condition, or the quality control substance is a rabbit lgG antibody, and the quality control capture antibody is a goat anti-rabbit lgG antibody.
A box cover of the reagent box body 7 is provided with a sample adding port 8 corresponding to the sample pad 5 so that a sample to be detected is added into the box cavity and falls on the sample pad 5. The box cover is further provided with a display window 9 at a position corresponding to the detection region and the quality control region, and distinguishing marks corresponding to the detection region and the quality control region one by one can be written beside the display window 9, as shown in fig. 2, the distinguishing mark of the quality control region is C, and the detecting distinguishing mark of the novel coronavirus (2019-nCoV) is T1.
When the sample dilution device is used, a sample to be detected is dripped from the sample adding port 8 to the sample pad 5, and then a sample dilution liquid is dripped for dilution, wherein the sample dilution liquid can be 0.01M phosphate buffer solution, and the sample can be a whole blood sample or extracted serum or plasma. The sample moves from the sample pad 5 to the water absorbing filter paper 1 in turn by capillary action, and is filtered by the blood filter pad 4, then flows through the combination pad 3, if the sample contains the novel coronavirus (2019-nCoV), the antigen will combine with the novel coronavirus (2019-nCoV) monoclonal detection antibody coated on the combination pad 3 to generate a compound, then the compound and the quality control substance on the combination pad 3 will flow continuously with the sample liquid, when the sample flows through the detection zone of the nitrocellulose membrane 2, the compound will combine with the novel coronavirus (2019-nCoV) monoclonal capture antibody specifically, because the novel coronavirus (2019-nCoV) monoclonal capture antibody is coated fixedly, the compound and the capture antibody will be captured and fixed and stay there, after reaching a certain amount, the detection zone will display the color of the label visible to the naked eye; when the sample flows through the quality control area, the quality control substance can be specifically combined with the quality control capture antibody and is captured in the quality control area in a concentrated manner, and after a certain amount of the sample flows through the quality control area, the color of the marker which can be seen by naked eyes can be displayed in the quality control area. If the detection area and the quality control area both show the color carried by the marker, as shown in FIG. 3, the sample shows to be positive for the novel coronavirus (2019-nCoV) carrying the novel coronavirus (2019-nCoV); if only the quality control region shows the color carried by the marker, as shown in fig. 6, the sample shows negative for the novel coronavirus (2019-nCoV) and does not carry the novel coronavirus (2019-nCoV); if neither the detection zone nor the quality control zone exhibits the color carried by the label, as shown in FIG. 2, the detection is disabled and needs to be re-detected.
With such a configuration, the novel coronavirus and influenza virus detection kit provided by the embodiment can simplify a detection process of a novel coronavirus (2019-nCoV), has no dependence on any experimental instrument, environment or operator, and is simple and convenient to operate, short in detection period, high in efficiency and easy to interpret; the kit needs a small amount of samples for detection, detects through antibody reaction, has high detection sensitivity, and has the advantages of low cost and the like; the detection process does not need special instruments and equipment, does not depend on higher laboratory conditions, detection personnel do not need professional training, the adaptability is strong, the monitoring and the detection at any time and any place are convenient, the rapid, efficient and low-cost diagnosis and detection are realized, and the method can be popularized in primary community hospitals and rural hospitals.
It should be noted that the above-mentioned detection process involves an immunoreaction principle, a labeling principle and means, which are conventional techniques, and antibody coating and capturing are also conventional techniques. For example, the detection area of the nitrocellulose membrane 2 is coated with the novel coronavirus monoclonal capture antibody by the prior art of hydrophobic interaction, hydrogen bond, electrostatic interaction and the like, and the quality control capture antibody is coated in the quality control area.
Meanwhile, influenza a virus is the most common influenza virus and one of the viruses which are easy to change, and is usually manifested as influenza-like symptoms, including fever, pharyngalgia, watery nasal discharge, nasal obstruction, cough, expectoration, headache, general aching pain and hypodynamia, and vomiting and diarrhea appear in some cases. Influenza b virus is a short for influenza virus and is an acute respiratory infectious disease with extremely strong infectivity caused by influenza virus. The virus B often causes localized epidemics, patients have symptoms of general malaise, high fever, myalgia and the like, and complications such as encephalitis, pneumonia and the like can occur when the disease condition is not controlled. The influenza A virus and the influenza B virus have infectivity and huge harmfulness, and the caused clinical symptoms and the caused complications have similarity with the novel coronavirus, so that the novel coronavirus, the influenza A virus and the influenza B virus are detected and distinguished, particularly, the effective distinction of virus types is carried out at the 2019-nCoV virus infection stage, the prevention and the treatment of the novel coronavirus and the panic reduction of epidemic situations are facilitated, and the diagnosis and the treatment of the novel coronavirus and the influenza A/B virus have guiding significance. Therefore, in this embodiment, the binding pad 3 of the detection kit is further coated with a monoclonal detection antibody for influenza a and/or b virus, and the detection region of the nitrocellulose membrane 2 is correspondingly coated and fixed with a monoclonal capture antibody for influenza a and/or b virus. The novel detection kit for coronavirus and influenza virus provided by the embodiment can be used for rapidly and efficiently detecting a sample, clinically distinguishing two or three viruses at a time, determining the types of the viruses carried in the sample, and rapidly distinguishing and diagnosing two or three diseases with clinical symptoms and complications at a time, so that the detection process is simplified, and the diagnosis confirming efficiency is improved.
Specifically, strip-shaped envelope lines are arranged at intervals along the detection area in the sample flowing direction, the novel coronavirus monoclonal capture antibody, the influenza a virus monoclonal capture antibody and the influenza b virus monoclonal capture antibody are respectively positioned on one of the envelope lines, the three envelope lines are in one-to-one correspondence with the three envelope lines, and are all coated and fixed on the corresponding envelope lines to form a novel coronavirus detection zone 11, an influenza a virus detection zone 12 and an influenza b virus detection zone 13. The quality control region coating the quality control capture antibody may be a coating line in the form of a line band, forming the quality control strip 10. The influenza A virus monoclonal capture antibody and the influenza B virus monoclonal capture antibody can be fixed and coated on the nitrocellulose membrane 2 through the prior art of hydrophobic interaction, hydrogen bond, electrostatic interaction and the like.
When the kit is used, a sample and diluent are added into the sample pad 5 from a sample port, the sample flows through the detection zone, and the complex carrying the corresponding viral antigen is bound to the capture antibody on the corresponding detection zone and is captured on the detection zone in a concentrated manner, so that the corresponding detection zone shows color. As shown in fig. 2 to 6, the three detection zones are written with distinguishing marks on the reagent cartridge body 7. The discrimination band of the novel coronavirus detection band is identified as T1, the discrimination band of the influenza A virus detection band 12 is identified as T2, and the discrimination band of the influenza B virus detection band is identified as T3. If the sample carries influenza A virus, the quality control strip 10 and the influenza A virus detection strip 12 can display colors, and the detection kit can be displayed as shown in FIG. 4; if the sample carries the influenza B virus, the quality control strip 10 and the influenza B virus detection strip 13 can display colors, and the detection kit can display as shown in figure 5; if the sample carries the novel coronavirus, the quality control strip 10 and the novel influenza virus detection strip 11 can display colors, and the detection kit can be displayed as shown in figure 3; if the sample does not carry the three types of viruses, only the quality control strip 10 will show the color, and the detection kit will be shown in FIG. 6. According to the arrangement, confusion can be avoided, detection results of virus types can be distinguished and displayed, and the virus type detection device is intuitive, convenient and efficient.
The color displayed in the display window 9 depends on the type of the label, and if the label carries a lanthanide or a chelate thereof such as samarium, europium or terbium, the color displayed is red or magenta. The display window 9 may be a large window covering the detection area and the quality control area, or may be a plurality of small windows corresponding to the quality control area and the three detection zones one to one. The cover of the reagent box body 1 is provided with an opening and a transparent object such as transparent plastic or glass for covering the opening to form the display window 9.
As shown in fig. 2, the novel coronavirus and influenza virus detection kit can be in the shape of a flat box, the sample addition port 8 is in the shape of a Chinese character hui, a display window 9 is arranged in the range of the box cover corresponding to the detection area and the quality control area, and the distinguishing mark is positioned on one side of the display window 9.
The following description of the kit is given by taking specific numerical values as examples: the length of the sample pad 5 is 14.0mm, the length of the blood filter pad 4 is 11.0mm, and the overlapping length of the sample pad 5 and the blood filter pad 4 which are overlapped and stuck together is 1.5 mm. The top end of the blood filtering pad 4 presses and is adhered to the bottom end of the combination pad 3, the length of the combination pad 3 is 6.0mm, the overlapping length of the blood filtering pad 4 and the combination pad 3 which are overlapped and adhered together is 1.5mm, the top end of the combination pad 3 presses and is adhered to the bottom end of the nitrocellulose membrane 2, and the overlapping length of the combination pad 3 and the nitrocellulose membrane 2 which are overlapped and adhered together is 2.0 mm. The top end of the cellulose nitrate film 2 is pressed and stuck with the bottom end of the water absorption filter paper 1, the length of the water absorption filter paper 1 is 32.0mm, and the overlapping length of the water absorption filter paper 1 and the cellulose nitrate film 2 which are overlapped and stuck together is 2.0 mm. The final concentration of the label is 0.5-1.0mg/ml, and the diameter of the label is 100nm-500 nm. The final concentration of the novel coronavirus monoclonal capture antibody and the influenza A/B virus monoclonal capture antibody for identifying the single epitope is 1-1.5 mg/ml; the final concentration of the quality control capture antibody is 0.5-1.0mg/ml, and the coating amount is 0.01-5 mug/cm. When the novel detection kit for coronavirus and influenza virus provided by the embodiment is used for detection, 10 mul of serum or whole blood sample is absorbed by a pipette gun and directly dripped into the sample adding port 8, and then 70-100 mul of sample diluent is dripped. Only 15 minutes are required to read the results.
It is understood that the same or similar parts in the above embodiments may be mutually referred to, and the same or similar parts in other embodiments may be referred to for the content which is not described in detail in some embodiments. The utility model provides a plurality of schemes contain the basic scheme of itself, mutual independence to restrict each other, but it also can combine each other under the condition of not conflicting, reaches a plurality of effects and realizes jointly.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.