Dengue virus NS1 antigen quantum dot fluorescence immunochromatography detection test strip and detection card

Abstract

The utility model relates to a dengue virus NS1 antigen quantum dot fluorescence immunochromatography detection test strip and a detection card. The dengue virus NS1 antigen quantum dot fluorescence immunochromatography detection test strip comprises a substrate, a sample pad, a first quantum dot labeling pad, a second quantum dot labeling pad, a first nitrocellulose membrane, a second nitrocellulose membrane, first absorbent paper and second absorbent paper, the dengue virus NS1 antigen quantum dot fluorescence immunochromatography detection test strip and the detection card are relatively good in dengue virus detection applicability, good in detection effect and convenient to use.

Description

Technical Field

[0001] This utility model belongs to the field of test strip manufacturing technology, specifically relating to a dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card. Background Technology

[0002] CN 205679623 U, entitled "A rapid detection test strip and kit for dengue virus NS1 antigen colloidal gold", describes a test strip comprising a substrate support layer and a reaction reagent carrier adsorption layer in the height direction. The reaction reagent carrier adsorption layer is fixed on the substrate support layer. From the handheld end to the sample application end, the strip includes a handheld absorbent pad, a nitrocellulose drug film, a gold-labeled fiber pad, and a sample fiber pad. A protective film layer covers the gold-labeled fiber pad and the handheld absorbent pad. A strip-shaped detection line and a control line perpendicular to the length of the test strip are printed on the nitrocellulose drug film. From the handheld end to the sample application end, the nitrocellulose drug film is sequentially coated with a control line (C line) and a detection line (T line) for detecting dengue NS1 antigen. The gold-labeled fiber pad of the test strip is coated with colloidal gold-labeled anti-dengue NS1 monoclonal antibody 2, the detection line (T line) is coated with anti-dengue NS1 monoclonal antibody 1, and the control line (C line) is coated with anti-mouse IgG polyclonal antibody. The gold-labeled fiber pad is coated with colloidal gold-labeled anti-dengue NS1 monoclonal antibody 2 at a concentration of 10-20 μg / ml. The test line T is coated with anti-dengue NS1 monoclonal antibody 1 at a concentration of 1.0-2.0 mg / ml, and the control line C is coated with anti-mouse IgG polyclonal antibody at a concentration of 1.0-2.0 mg / ml. A rapid detection kit for dengue virus NS1 antigen using colloidal gold includes the aforementioned colloidal gold test strip. It also includes a sealed box consisting of a box body and a lid, the box body having a positioning groove for mounting the colloidal gold test strip. The lid has a sample application hole and a result observation hole; when the colloidal gold test strip is placed in the positioning groove within the box body, the sample fiber pad corresponds to the sample application hole on the lid, and the nitrocellulose drug membrane corresponds to the result observation hole. The kit is rectangular, elliptical, semi-circular, or rounded rectangular. The current rapid dengue virus NS1 antigen colloidal gold test strip and kit can only perform rapid qualitative detection of dengue virus (with relatively poor applicability); therefore, our company has developed a dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card that can perform quantitative detection of dengue virus (with relatively better applicability). Utility Model Content

[0003] Design objective: To overcome the shortcomings of the prior art, this paper designs a dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card that is not only more suitable for dengue virus detection, but also has good detection effect and is easy to use.

[0004] Design scheme: To achieve the above design objectives.

[0005] 1. A sample pad is provided on the upper surface of the substrate, and the sample pad is located in the center of the substrate. A first quantum dot labeling pad and a second quantum dot labeling pad are provided on the upper surface of the substrate, and the first quantum dot labeling pad and the second quantum dot labeling pad are located on both sides of the sample pad. The lower end face of one end of the sample pad overlaps with the upper end face of one end of the first quantum dot labeling pad, and the lower end face of the other end of the sample pad overlaps with the upper end face of one end of the second quantum dot labeling pad. A first nitrocellulose membrane is provided on the upper surface of the substrate, and the upper end face of one end of the first nitrocellulose membrane overlaps with the lower end face of the other end of the first quantum dot labeling pad. The substrate has a second nitrocellulose membrane on its upper surface, with the upper end face of one end of the second nitrocellulose membrane overlapping the lower end face of the other end of the second quantum dot labeling pad. The substrate also has a first absorbent paper on its upper surface, with the lower end face of one end of the first absorbent paper overlapping the upper end face of the other end of the first nitrocellulose membrane. The design of having a detection line and a quality control line on the first nitrocellulose membrane, and a reference line on the second nitrocellulose membrane, is one of the technical features of this invention.The purpose of this design is as follows: a sample pad is provided on the upper surface of the substrate, and the sample pad is located in the exact center of the substrate. A first quantum dot labeling pad and a second quantum dot labeling pad are provided on the upper surface of the substrate, and the first and second quantum dot labeling pads are located on opposite sides of the sample pad. The lower end face of one end of the sample pad overlaps with the upper end face of one end of the first quantum dot labeling pad, and the lower end face of the other end of the sample pad overlaps with the upper end face of one end of the second quantum dot labeling pad. A first nitrocellulose filament is provided on the upper surface of the substrate. The substrate has a first nitrocellulose membrane, with the upper end face of one end overlapping the lower end face of the other end of the first quantum dot labeling pad. The substrate also has a second nitrocellulose membrane on its upper surface, with the upper end face of one end overlapping the lower end face of the other end of the second quantum dot labeling pad. Furthermore, the substrate has a first absorbent paper on its upper surface, with the lower end face of one end overlapping the upper end face of the other end of the first nitrocellulose membrane. Finally, the substrate has a second absorbent paper on its upper surface, with one end of the second absorbent paper overlapping the lower end face of the other end of the first nitrocellulose membrane. The lower end face of the side end overlaps with the upper end face of the other side end of the second nitrocellulose membrane. The first nitrocellulose membrane has a detection line and a control line, while the second nitrocellulose membrane has a reference line. This bidirectional test strip, with the sample pad in the middle, features a detection line and control line on one side and a reference line on the other. This structure allows for quantitative detection of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card by reading the fluorescence intensity value at the reference line (not only quickly determining whether dengue virus infection has occurred (qualitative), but also quickly determining the strength and weakness of the infection (quantitative)). This improves the applicability of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card. Simultaneously, the reference line is far from the detection line and control line, avoiding mutual interference during fluorescence intensity readings and improving detection efficiency. Furthermore, the sample pad's central location makes the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card simpler and more convenient to use.

[0006] 2. The design that the control line and reference line are kept at a consistent distance from the sample pad, and the distance between the detection line and the control line is 6mm-10mm, is the second technical feature of this utility model. The purpose of this design is to ensure that the control line and reference line are kept at a consistent distance from the sample pad, and the distance between the detection line and the control line is 6mm-10mm, thus further ensuring the detection effect of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card.

[0007] 3. The upper end face of the upper shell of the plastic card case is provided with a first observation window, a second observation window, and a sample application hole, all of which penetrate the upper and lower end faces of the upper shell. The sample application hole is located between the first and second observation windows and directly above the sample pad. The first observation window is located directly above the first nitrocellulose membrane, and the second observation window is located directly above the second nitrocellulose membrane. This design is the third technical feature of this utility model. The purpose of this design is that the upper end face of the upper shell of the plastic card case is provided with a first observation window, a second observation window, and a sample application hole, all of which penetrate the upper and lower end faces of the upper shell. The sample application hole is located between the first and second observation windows and directly above the sample pad. The first observation window is located directly above the first nitrocellulose membrane, and the second observation window is located directly above the second nitrocellulose membrane, thus facilitating the observation of the test card.

[0008] Technical Solution 1: A dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip, comprising a substrate, a sample pad, a first quantum dot labeled pad, a second quantum dot labeled pad, a first nitrocellulose membrane, a second nitrocellulose membrane, a first absorbent paper, and a second absorbent paper. The sample pad is located at the center of the substrate on its upper surface. The first and second quantum dot labeled pads are located on opposite sides of the sample pad. The lower end of one end of the sample pad overlaps with the upper end of one end of the first quantum dot labeled pad, and the lower end of the other end of the sample pad overlaps with the upper end of one end of the second quantum dot labeled pad. The upper surface of the substrate is provided with a first quantum dot labeled pad, a second quantum dot labeled pad, a first nitrocellulose membrane, a second nitrocellulose membrane, a first absorbent paper, and a second absorbent paper. The substrate has a first nitrocellulose membrane, with the upper end of one end of the first nitrocellulose membrane overlapping the lower end of the other end of the first quantum dot labeling pad. The substrate also has a second nitrocellulose membrane on its upper surface, with the upper end of one end of the second nitrocellulose membrane overlapping the lower end of the other end of the second quantum dot labeling pad. A first absorbent paper is also provided on the upper surface of the substrate, with the lower end of one end of the first absorbent paper overlapping the upper end of the other end of the first nitrocellulose membrane. A second absorbent paper is also provided on the upper surface of the substrate, with the lower end of one end of the second absorbent paper overlapping the upper end of the other end of the second nitrocellulose membrane. The first nitrocellulose membrane has a detection line and a control line, and the second nitrocellulose membrane has a reference line.

[0009] Technical Solution 2: A test card, comprising a plastic card shell and a test strip, wherein the test strip is disposed within the plastic card shell, and the test strip is any of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strips described above.

[0010] Compared with the prior art, the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card of this invention are not only more suitable for dengue virus detection, but also have better detection effect and are more convenient to use. Attached Figure Description

[0011] Figure 1 This is a schematic diagram of the main structure of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip.

[0012] Figure 2 This is a top view of the test card.

[0013] Figure 3 It is the main view structural design drawing of the test card provided by the applicant.

[0014] Figure 4 This is the main structural design draft of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip provided by the applicant. Detailed Implementation

[0015] Example 1: Refer to Appendix Figures 1-4 A quantum dot fluorescent immunochromatographic test strip for dengue virus NS1 antigen includes a substrate 1, a sample pad 2, a first quantum dot labeled pad 3, a second quantum dot labeled pad 4, a first nitrocellulose membrane 5, a second nitrocellulose membrane 6, a first absorbent paper 7, and a second absorbent paper 8. The sample pad 2 is located at the center of the upper surface of the substrate 1. The first quantum dot labeled pad 3 and the second quantum dot labeled pad 4 are located on opposite sides of the sample pad 2. The lower end of one end of the sample pad 2 overlaps with the upper end of one end of the first quantum dot labeled pad 3, and the lower end of the other end of the sample pad 2 overlaps with the upper end of one end of the second quantum dot labeled pad 4. The upper surface of the substrate 1 is provided with a first nitrocellulose membrane 5, a first nitrocellulose membrane 6, a second nitrocellulose membrane 7, and a second absorbent paper 8. The substrate 1 has a second nitrocellulose membrane 6 on its upper surface, with one end of the second nitrocellulose membrane 5 overlapping the lower end of the other end of the first quantum dot labeling pad 3. The substrate 1 also has a first absorbent paper 7 on its upper surface, with one end of the first absorbent paper 7 overlapping the upper end of the other end of the first nitrocellulose membrane 5. The substrate 1 also has a second absorbent paper 8 on its upper surface, with one end of the second absorbent paper 8 overlapping the upper end of the other end of the second nitrocellulose membrane 6. The first nitrocellulose membrane 5 has a detection line 51 and a quality control line 52, and the second nitrocellulose membrane 6 has a reference line 61.

[0016] The control line 52 and the reference line 61 are kept at the same distance from the sample pad 2, and the distance between the detection line 51 and the control line 52 is 6mm-10mm. Preferably, the distance between the detection line 51 and the control line 52 is 8mm.

[0017] The detection line 51 is coated with dengue virus NS1 protein antibody, the control line 52 is coated with IgG antibody, and the reference line 61 is coated with IgG antibody. The first quantum dot labeled pad 3 is coated with dengue virus NS1 protein antibody labeled with quantum dot fluorescent microspheres, and the second quantum dot labeled pad 4 is coated with dengue virus NS1 protein antibody labeled with quantum dot fluorescent microspheres.

[0018] The substrate 1 is a PVC board, the first nitrocellulose membrane 5 and the second nitrocellulose membrane 6 are both German Sartorius CN140, the sample pad 2 is a Finnish Ahlstrom 8964 glass cellulose membrane, the first quantum dot labeling pad 3 and the second quantum dot labeling pad 4 are Finnish Ahlstrom 6613 polyester fiber membranes, and the first absorbent paper 7 and the second absorbent paper 8 are cellulose filter papers.

[0019] The first quantum dot labeled pad 3 and the second quantum dot labeled pad 4 are labeled with quantum dot fluorescent microspheres conjugated with dengue virus NS1 protein antibodies (commercially available products). (That is, the dengue virus NS1 protein antibodies labeled with quantum dot fluorescent microspheres are prior art products.) Preferably, the dengue virus NS1 protein antibody on the quantum dot labeled pad is any one of mouse monoclonal antibody, rabbit monoclonal antibody, or goat monoclonal antibody, which can recognize the NS1 protein of four serotypes of dengue virus. Preferably, the quantum dot fluorescent microspheres are any one of polymer microspheres or silica microspheres, with a size of 100~300 nm.

[0020] The detection line 51 is coated with a dengue virus NS1 protein antibody (a commercially available product, i.e., a prior art product). Preferably, the dengue virus NS1 protein antibody on the detection line 51 is any one of mouse monoclonal antibody, rabbit monoclonal antibody, or goat monoclonal antibody. It can be a single monoclonal antibody that can recognize four serotypes of NS1 protein simultaneously, or it can be a combination of monoclonal antibodies that can recognize one or more of the four serotypes of dengue virus NS1 protein, enabling the recognition and capture of the four serotypes of NS1 protein.

[0021] The control line 52 and the reference line 61 are coated with IgG antibodies (commercially available products, i.e., prior art products). Preferably, the IgG antibodies coated on the control line 52 and the reference line 61 are antibodies used in quantum dot labeled pads, and can be either monoclonal antibodies or polyclonal antibodies.

[0022] The steps for using the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip and test card are as follows:

[0023] (1) Use blood collection tubes or fingertip blood collection needles to collect venous or fingertip blood samples from patients. Take 20uL of the collected blood sample, add 180uL of sample diluent, and mix thoroughly by inverting the tubes three times.

[0024] (2) Take 3 drops (about 100 μL) of sample dilution solution and put it onto the sample pad 2 of the test strip;

[0025] (3) Let stand for 10-15 minutes, then observe the results under ultraviolet light or detect them with a dry immunofluorescence analyzer.

[0026] The sample dilution solution in step (1) above consists of PBS buffer, casein, bovine serum albumin and Tween-20, wherein the molar concentration of PBS buffer is 10 mM, the mass concentration of casein is 0.1%, the mass concentration of bovine serum albumin is 0.1%, and the mass concentration of Tween-20 is 0.5%.

[0027] Example 2: Based on Example 1. A test card includes a plastic card housing 100 and a test strip 200. The test strip 200 is disposed inside the plastic card housing 100, and the test strip 200 is any of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strips described above. The upper end face of the upper shell of the plastic card housing 100 is provided with a first observation window 101, a second observation window 102, and a sample application hole 103, and the first observation window 101, the second observation window 102, and the sample application hole 103 all penetrate the upper and lower end faces of the upper shell. The sample application hole 103 is located between the first observation window 101 and the second observation window 102, and is located directly above the sample pad 2. The first observation window 101 is located directly above the first nitrocellulose membrane 5, and the second observation window 102 is located directly above the second nitrocellulose membrane 6.

[0028] It should be understood that although the above embodiments provide a relatively detailed textual description of the design concept of this utility model, these textual descriptions are merely simple textual descriptions of the design concept of this utility model, and not limitations on the design concept of this utility model. Any combination, addition, or modification that does not exceed the design concept of this utility model shall fall within the protection scope of this utility model.

Claims

1. A quantum dot fluorescent immunochromatographic test strip for dengue virus NS1 antigen, comprising a substrate (1), a sample pad (2), a first quantum dot labeled pad (3), a second quantum dot labeled pad (4), a first nitrocellulose membrane (5), a second nitrocellulose membrane (6), a first absorbent paper (7), and a second absorbent paper (8), characterized in that: The upper surface of the substrate (1) is provided with a sample pad (2) and the sample pad (2) is located in the middle of the substrate (1). The upper surface of the substrate (1) is provided with a first quantum dot labeling pad (3) and a second quantum dot labeling pad (4) and the first quantum dot labeling pad (3) and the second quantum dot labeling pad (4) are located on both sides of the sample pad (2). The lower end face of one end of the sample pad (2) overlaps with the upper end face of one end of the first quantum dot labeling pad (3) and the lower end face of the other end of the sample pad (2) overlaps with the upper end face of one end of the second quantum dot labeling pad (4). The upper surface of the substrate (1) is provided with a first nitrocellulose membrane (5) and the upper end face of one end of the first nitrocellulose membrane (5) overlaps with the lower end face of the other end of the first quantum dot labeling pad (3). Next, the upper surface of the substrate (1) is provided with a second nitrocellulose membrane (6), and the upper surface of one end of the second nitrocellulose membrane (6) overlaps with the lower surface of the other end of the second quantum dot labeling pad (4). The upper surface of the substrate (1) is provided with a first absorbent paper (7), and the lower surface of one end of the first absorbent paper (7) overlaps with the upper surface of the other end of the first nitrocellulose membrane (5). The upper surface of the substrate (1) is provided with a second absorbent paper (8), and the lower surface of one end of the second absorbent paper (8) overlaps with the upper surface of the other end of the second nitrocellulose membrane (6). The first nitrocellulose membrane (5) is provided with a detection line (51) and a quality control line (52), and the second nitrocellulose membrane (6) is provided with a reference line (61).

2. The dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip according to claim 1, characterized in that: The distance between the control line (52) and the reference line (61) and the sample pad (2) is consistent, and the distance between the detection line (51) and the control line (52) is 6mm-10mm.

3. The dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip according to claim 2, characterized in that: The distance between the detection line (51) and the quality control line (52) is 8 mm.

4. The dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip according to claim 1 or 2, characterized in that: The detection line (51) is coated with dengue virus NS1 protein antibody, the control line (52) is coated with IgG antibody, and the reference line (61) is coated with IgG antibody.

5. The dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip according to claim 1 or 2, characterized in that: The first quantum dot labeling pad (3) is coated with dengue virus NS1 protein antibody labeled with quantum dot fluorescent microspheres, and the second quantum dot labeling pad (4) is coated with dengue virus NS1 protein antibody labeled with quantum dot fluorescent microspheres.

6. The dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strip according to claim 1, characterized in that: The substrate (1) is a PVC board, the first nitrocellulose membrane (5) and the second nitrocellulose membrane (6) are both German Sartorius CN140, the sample pad (2) is Finnish Ahlstrom 8964 glass cellulose membrane, the first quantum dot labeling pad (3) and the second quantum dot labeling pad (4) are Finnish Ahlstrom 6613 polyester fiber membrane, and the first absorbent paper (7) and the second absorbent paper (8) are cellulose filter paper.

7. A test card, comprising a plastic casing (100) and a test strip (200), wherein the test strip (200) is disposed within the plastic casing (100), characterized in that: The test strip (200) is any one of the dengue virus NS1 antigen quantum dot fluorescent immunochromatographic test strips as described in claims 1-6.

8. The detection card according to claim 7, characterized in that: The upper end face of the upper shell of the plastic card holder (100) is provided with a first observation window (101), a second observation window (102) and a sample application hole (103), and the first observation window (101), the second observation window (102) and the sample application hole (103) all penetrate the upper and lower end faces of the upper shell. The sample application hole (103) is located between the first observation window (101) and the second observation window (102). The sample application hole (103) is located directly above the sample pad (2). The first observation window (101) is located directly above the first nitrocellulose membrane (5), and the second observation window (102) is located directly above the second nitrocellulose membrane (6).

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