CN212989383U - Coronavirus antibody joint inspection test paper strip and reagent card - Google Patents
Coronavirus antibody joint inspection test paper strip and reagent card Download PDFInfo
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- CN212989383U CN212989383U CN202020713371.5U CN202020713371U CN212989383U CN 212989383 U CN212989383 U CN 212989383U CN 202020713371 U CN202020713371 U CN 202020713371U CN 212989383 U CN212989383 U CN 212989383U
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Abstract
The utility model relates to a coronavirus antibody joint inspection test paper strip and a reagent card, which are formed by sticking a PVC bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad; the reagent for detecting the novel coronavirus antibody by the colloidal gold chromatography can judge the result in 10 minutes, thereby preventing epidemic situation from spreading as soon as possible.
Description
Technical Field
The utility model relates to a biological application technical field especially relates to a coronavirus antibody joint inspection test paper strip.
Background
The novel coronavirus SARS-CoV-2 is a closely related coronavirus causing COVID-19, patients with COVID-19 mainly show fever, dry cough and hypodynamia, a few patients have symptoms such as nasal obstruction, watery nasal discharge, pharyngalgia, myalgia, diarrhea and the like, severe patients often have dyspnea and/or hypoxemia after one week of morbidity, and severe patients can rapidly progress to acute respiratory distress syndrome, septic shock, metabolic acidosis which is difficult to correct, blood coagulation dysfunction, multiple organ failure and the like.
The novel coronavirus has strong concealment, and patients with unobvious symptoms or even patients with asymptomatic infection. New coronaviruses, which are currently spreading exponentially in more than 200 countries worldwide, are posing serious challenges to global health care systems and institutions.
Disclosure of Invention
The utility model aims at providing a coronavirus antibody joint inspection test paper strip has advantages such as easy and simple to handle, the reaction is quick, the sensitivity is high, the specificity is strong, be fit for witnessed inspections. The rapid preliminary screening of the febrile patients can improve the diagnosis rate of suspected cases, for example, the large-scale screening of asymptomatic people is realized, and the IgM antibody and the IgG antibody in the human body are simultaneously detected, so that the rapid detection of potential coronavirus infectors is facilitated.
The utility model provides a coronavirus antibody joint inspection test paper strip, includes the PVC bottom plate, is stained with sample pad, combination pad, nitrocellulose membrane and absorbent pad, its characterized in that in the one side of PVC bottom plate in order each other take up: the combined pad is a glass cellulose film coated with a recombinant novel coronavirus antigen-colloidal gold compound and a chicken IgY antibody-colloidal gold compound, and the nitrocellulose film is respectively coated with a detection line of a mouse anti-human IgM antibody, a detection line of a mouse anti-human IgG antibody and a quality control line of a goat anti-chicken IgY antibody.
The coronavirus antibody joint inspection test strip is characterized in that: spraying the mixed solution of the recombinant novel coronavirus antigen-colloidal gold compound and the chicken IgY antibody-colloidal gold compound onto the binding pad, and drying in an oven at 37 ℃ for 12-24 h.
The coronavirus antibody joint inspection test strip is characterized in that: the mouse anti-human IgG antibody, the mouse anti-human IgM antibody and the goat anti-chicken IgY antibody are obtained by diluting with an antibody diluent, coating on a nitrocellulose membrane, and drying at 37 ℃ for 4 hours.
The coronavirus antibody joint inspection test strip is characterized in that: the antigen is selected from one or two of recombinant novel coronavirus S protein and recombinant novel coronavirus N protein.
The coronavirus antibody joint inspection test strip is characterized in that: the antibody diluent is prepared by dissolving sucrose in phosphate buffer solution with pH 7.4.
A reagent card for combined detection of coronavirus antibodies, which is characterized in that: the reagent card comprises the detection test strip and a reagent card shell.
A reagent card box for combined detection of coronavirus antibody comprises a sample diluent and the detection reagent card.
The reagent card box for the combined detection of the coronavirus antibody is characterized in that: the sample diluent is phosphate buffer solution with pH of 7.0 +/-0.2.
A preparation method of a coronavirus antibody joint inspection test strip is characterized by comprising the following steps:
1) preparation of a marking solution: adding 1.0ml of 10% chloroauric acid solution into 1000ml of boiling water, stirring while adding, fully boiling for 10 seconds, adding 1.8ml of 10% sodium citrate solution, keeping heating, reacting for 30 minutes, stopping heating, and continuously stirring to room temperature;
2) preparing a labeled recombinant novel coronavirus antigen solution: 100ml of the marking solution is added dropwise with 0.05M K2CO35.0ml (pH is about 8.0), fully mixing, dropwise adding 0.1mg/ml recombinant novel coronavirus antigen aqueous solution, fully mixing for 30min, dropwise adding 13ml 10% bovine serum albumin solution, slowly stirring for 30min, centrifuging at low temperature and high speed for 30min, removing supernatant, and dissolving precipitate with colloidal gold diluent to obtain the final product;
3) preparation of labeled antibody solution: 0.05M K was added dropwise to 100ml of the labeling solution2CO35.0ml (pH about 8.0), mixing thoroughly, adding 0.1mg/ml chicken IgY antibody water solution dropwise, mixing thoroughly for 30min, adding 10% bovine serum dropwiseSlowly stirring 13ml of protein solution for 30min, centrifuging at low temperature and high speed for 30min, removing supernatant, and dissolving precipitate with colloidal gold diluent to obtain the final product;
4) preparing an antibody coating solution: dissolving 2g of sucrose in 100ml of 0.2M phosphate buffer solution with pH7.4, filtering with a 0.22 mu M pore size microporous filter membrane, and storing at 2-8 ℃;
5) c, T preparation of coating solution: diluting goat anti-chicken IgY antibody, mouse anti-human IgM antibody and mouse anti-human IgG to 0.3-0.5mg/ml, 0.5-0.8mg/ml and 0.5-0.8mg/ml respectively by using antibody coating solution;
6) labeling with a glass cellulose membrane: mixing and spraying the marked recombinant coronavirus antigen solution and the marked antibody solution onto the cut bonding pad according to the ratio of 3:1-4:1, wherein the liquid spraying amount is 4-5 mu l/cm, and drying in an oven at 37 ℃ for 12-24 hours;
7) and (3) carrying out scribing treatment on the nitrocellulose membrane: scribing C, T line packets on the nitrocellulose membrane by a gold spraying membrane scribing instrument according to 1 mul/cm, and drying the NC membrane in an oven at 37 ℃ for 12-24h after scribing is finished;
8) preparing a test strip: in a drying chamber, taking a PVC base plate, and sequentially overlapping and adhering a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad on one surface of the PVC base plate in an environment with the temperature of 18-28 ℃ and the humidity of less than 30%; and cutting the adhered PVC base plate into test strips with the width of more than or equal to 2.5mm by using a cutting machine.
Preparing a coronavirus antibody joint detection reagent card: and (3) loading the test strip into a clamping position at the bottom of the detection card, aligning the sample adding hole of the card cover with the sample pad, aligning the observation window with the detection line and the quality control line, and assembling into a reagent card.
Compared with the prior art, the utility model have following advantage:
1. the diagnosis is rapid, and the detection can be completed within 10 minutes; 2. the IgM antibody and the IgG antibody in the human body can be simultaneously and respectively detected only by adding the sample once; 3. the operation is simple and convenient, and the operation by professional personnel is not needed; 4. the blood sample that serology detected is changeed and is acquireed and sample quality is guaranteed, has reduced medical personnel's risk of being infected at sample collection and testing process to a great extent.
Drawings
The above is an overview of the technical solution of the present invention, and in order to further explain the present invention, the following detailed description is made in conjunction with the accompanying drawings and the embodiments.
FIG. 1 is a schematic structural diagram of a coronavirus antibody joint inspection test strip of the present invention.
FIG. 2 is a schematic view of the structure of a reagent card cover for joint inspection of coronavirus antibodies of the present invention
FIG. 3 is a schematic view of the structure of the reagent card bottom for combined detection of coronavirus and antibody of the present invention
Description of the symbols of the drawings: the detection kit comprises a detection line 1-IgM, a detection line 2-IgG, a detection line 3-quality control line, a sample pad 4-5-combination pad, a nitrocellulose membrane 6-7-PVC bottom plate, a water absorption pad 8-9-sample adding holes 10-observation windows 11-one end of a detection test strip clamping position and the other end of the detection test strip clamping position.
Detailed Description
Example 1: coronavirus antibody joint detection test strip, reagent card preparation and detection method
Preparation of a marking solution: adding 10% chloroauric acid solution 1.0ml into 1000ml boiling water, stirring while adding, boiling for 10 seconds, adding 10% sodium citrate solution 1.8ml, maintaining heating, reacting for 30 minutes, stopping heating, and stirring to room temperature.
Preparing a labeled recombinant novel coronavirus antigen solution: 100ml of the marking solution is added dropwise with 0.05M K2CO35.0ml (pH is about 8.0), mixing completely, adding 0.1mg/ml recombinant novel coronavirus antigen aqueous solution dropwise, mixing completely for 30min, adding 13ml 10% bovine serum albumin solution dropwise, stirring slowly for 30min, centrifuging at low temperature and high speed for 30min, discarding supernatant, and dissolving precipitate with colloidal gold diluent to obtain the final product.
Preparation of labeled antibody solution: 0.05M K was added dropwise to 100ml of the labeling solution2CO35.0ml (pH about 8.0), mixing, dropwise adding 0.1mg/ml chicken IgY antibody water solution, mixing for 30min, dropwise adding 10% bovine serum albumin solution 13ml, stirring slowly for 30min, centrifuging at low temperature and high speed for 30min, discarding supernatant, precipitating with colloidal gold diluent solutionAnd (5) obtaining the product after solution.
Preparing an antibody coating solution: dissolving 2g of sucrose in 100ml of 0.2MpH7.4 phosphate buffer solution, filtering with a 0.22 μm pore size microporous filter membrane, and storing at 2-8 deg.C.
C. Preparing a T-line coating solution: the goat anti-chicken IgY antibody, the mouse anti-human IgM antibody, the mouse anti-human IgG are respectively diluted to 0.3mg/ml, 0.5mg/ml and 0.5mg/ml by using the antibody coating solution.
Labeling with a glass cellulose membrane: and mixing the marked recombinant coronavirus antigen solution and the marked antibody solution according to the ratio of 3:1, spraying onto the cut bonding pad, spraying the solution with the volume of 5 mu l/cm, and drying in an oven at 37 ℃ for 12 h.
And (3) carrying out scribing treatment on the nitrocellulose membrane: the C, T wire wrap solution was scribed on the nitrocellulose membrane using a gold spraying scriber at 1. mu.l/cm, and after scribing was complete the NC membrane was dried in an oven at 37 ℃ for 12 h.
Assembling a reagent card: in a drying chamber, taking a PVC base plate, and sequentially overlapping and adhering a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad on one surface of the PVC base plate in an environment with the temperature of 18-28 ℃ and the humidity of less than 30%; cutting the adhered PVC base plate into test strips with the width of 2.5mm by a cutting machine; then the test card cover and the test card bottom are assembled into a reagent card, wherein the sample adding hole of the card cover is aligned with the sample pad, and the observation window is aligned with the detection line and the quality control line.
The detection method comprises the following steps:
1. taking out the sample to be tested and the required reagent from the storage condition, and horizontally placing;
2. the sample is fully and evenly mixed before testing, 10 mul of sample to be tested (serum, plasma or whole blood sample) is added into a sample hole on a detection card, and 2 drops of sample diluent are immediately dropped into the sample hole.
And (3) detection results: 55 clinical serum of patients with confirmed diagnosis of the novel coronavirus pneumonia and 55 serum of patients with exclusion of the novel coronavirus pneumonia are detected, the sensitivity of the result is 96.4%, and the specificity is 98.2%. The detection result shows that the prepared detection reagent has good performance and is beneficial to quickly detecting a person infected by the novel coronavirus.
Example 2: preparation of coronavirus antibody joint inspection test strip and reagent card
Preparation of a marking solution: 1.0mL of 10% chloroauric acid solution was added to 1000mL of boiling water, the mixture was stirred while boiling for 10 seconds, 1.8mL of 10% sodium citrate solution was added thereto, the heating was maintained, the reaction was continued for 30 minutes, the heating was stopped, and the mixture was stirred to room temperature.
Preparing a labeled recombinant novel coronavirus antigen solution: 100ml of the marking solution is added dropwise with 0.05M K2CO35.0ml (pH is about 8.0), mixing completely, adding 0.1mg/ml recombinant novel coronavirus antigen aqueous solution dropwise, mixing completely for 30min, adding 13ml 10% bovine serum albumin solution dropwise, stirring slowly for 30min, centrifuging at low temperature and high speed for 30min, discarding supernatant, and dissolving precipitate with colloidal gold diluent to obtain the final product.
Preparation of labeled antibody solution: 0.05M K was added dropwise to 100ml of the labeling solution2CO35.0ml (pH is about 8.0), mixing completely, adding 0.1mg/ml chicken IgY antibody water solution dropwise, mixing completely for 30min, adding 13ml 10% bovine serum albumin solution dropwise, stirring slowly for 30min, centrifuging at low temperature and high speed for 30min, discarding supernatant, and dissolving precipitate with colloidal gold diluent to obtain the final product.
Preparing an antibody coating solution: dissolving 2g of sucrose in 100ml of 0.2MpH7.4 phosphate buffer solution, filtering with a 0.22 μm pore size microporous filter membrane, and storing at 2-8 deg.C.
C. Preparing a T-line coating solution: the goat anti-chicken IgY antibody, the mouse anti-human IgM antibody, the mouse anti-human IgG are respectively diluted to 0.5mg/ml, 0.8mg/ml and 0.8mg/ml by using the antibody coating solution.
Labeling with a glass cellulose membrane: and mixing the marked recombinant coronavirus antigen solution and the marked antibody solution according to the ratio of 4:1, spraying onto the cut bonding pad, spraying at 4 mu L/cm, and drying in an oven at 37 ℃ for 24 h.
And (3) carrying out scribing treatment on the nitrocellulose membrane: the C, T wire wrap solution was scribed on the nitrocellulose membrane at 1. mu.L/cm using a gold spraying scriber, and after scribing was completed, the NC membrane was dried in an oven at 37 ℃ for 24 hours.
Assembling a reagent card: in a drying chamber, taking a PVC base plate, and sequentially overlapping and adhering a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad on one surface of the PVC base plate in an environment with the temperature of 18-28 ℃ and the humidity of less than 30%; cutting the adhered PVC base plate into test strips with the width of 3mm by using a cutting machine; then the test card cover and the test card bottom are assembled into a reagent card, wherein the sample adding hole of the card cover is aligned with the sample hole, and the observation window is aligned with the detection line and the quality control line.
Although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the present invention can be modified or replaced with other embodiments without departing from the spirit and scope of the present invention.
Claims (4)
1. The utility model provides a coronavirus antibody joint inspection test paper strip, includes the PVC bottom plate, is stained with sample pad, combination pad, nitrocellulose membrane and absorbent pad, its characterized in that in the one side of PVC bottom plate in order each other take up: the combined pad is a glass cellulose film coated with a recombinant novel coronavirus antigen-colloidal gold compound and a chicken IgY antibody-colloidal gold compound, and the nitrocellulose film is respectively coated with a detection line of a mouse anti-human IgM antibody, a detection line of a mouse anti-human IgG antibody and a quality control line of a goat anti-chicken IgY antibody.
2. The coronavirus antibody joint inspection test strip of claim 1, which is characterized in that: the antigen is selected from one or two of recombinant novel coronavirus S protein and recombinant novel coronavirus N protein.
3. A coronavirus antibody reagent card, comprising: the reagent card comprises a reagent card shell and the joint test strip of claim 1.
4. The coronavirus antibody reagent card of claim 3, wherein: the test paper strip is located the screens department at reagent card bottom, and the application of sample hole of card lid aligns the sample pad, and the inspection line and quality control line are aimed at to the observation window.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202020713371.5U CN212989383U (en) | 2020-05-06 | 2020-05-06 | Coronavirus antibody joint inspection test paper strip and reagent card |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202020713371.5U CN212989383U (en) | 2020-05-06 | 2020-05-06 | Coronavirus antibody joint inspection test paper strip and reagent card |
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| CN212989383U true CN212989383U (en) | 2021-04-16 |
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