Multi-joint test quantum dot quantitative test cassette, preparation method and device therefor, and use method therefor

Abstract

The present application belongs to the field of biological testing. Disclosed are a multi-joint test quantum dot quantitative test cassette, a preparation method and device therefor, and a use method therefor. The multi-joint test quantum dot quantitative test cassette comprises a multi-joint-row cassette shell and test strips, wherein a plurality of reaction cavities are provided in the multi-joint-row cassette shell and are used for the placement of the test strips; the multi-joint-row cassette shell further comprises a plurality of sample-adding holes and a plurality of observation windows, the sample-adding holes and the observation windows corresponding to the reaction cavities on a one-to-one basis; and each test strip comprises a binding pad, which is coated with an antibody marked by quantum dots, the antibody marked by the quantum dots serving as a chromogenic substance. The present application provides a multi-joint test quantum dot quantitative test cassette, which uses quantum-dot-marked quantitative testing, a nitrocellulose membrane with a plurality of test lines, and the form of the multi-joint-row cassette shell. Thus, the problems of quantitative testing and multi-joint testing are both solved, and the test efficiency and accuracy are improved; furthermore, the present application achieves the aim of rapidly diagnosing various allergic diseases of a human body that are caused by the inhalation and ingestion of allergens.

Description

Multi-detection quantum dot quantitative detection card, preparation method, device and use method

[0001] This application claims priority to the Chinese patent application filed with the Patent Office of China on December 26, 2022, with application number 202211682483.9 and titled “A multi-detection quantum dot quantitative detection card and its preparation method”, and the Chinese patent application filed with the Patent Office of China on December 26, 2022, with application number 202211676895.1 and titled “A multi-detection device and its use method”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present application relates to the field of biological detection, and in particular to a multi-detection quantum dot quantitative detection card, preparation method, device and use method. Background Art

[0003] The prevalence of allergic diseases is currently increasing. my country already has 300-400 million patients with allergic diseases, and the number is increasing annually. The diagnosis of allergic diseases primarily involves in vivo and in vitro allergen testing. In vitro testing is gaining increasing attention, particularly for pediatric patients, due to its safety, readily available preparations, lack of impact from medications, and ability to simultaneously detect multiple allergens. Allergen-specific IgE (sIgE) antibody testing plays a crucial role in reflecting the condition, assisting in diagnosis, and predicting the onset and prognosis of disease, and is incorporated into the diagnostic and treatment guidelines for numerous clinical conditions. IgE test results can be categorized as either quantitative or qualitative. Qualitative results, such as immunoblotting, simply indicate a negative or positive result. Fully quantitative allergen testing, on the other hand, measures the specific level of IgE antibodies in serum, providing more intuitive numerical results that reflect disease relevance and more effectively predict disease onset and prognosis. This makes it more clinically meaningful for diagnosing allergic diseases. Quantitative testing offers superior precision and reproducibility, resulting in more reliable results.

[0004] Existing allergen-specific quantitative detection methods, such as enzyme-linked immunosorbent assays (ELISAs), are time-consuming, complex, and require large-scale equipment. Therefore, fluorescent immunoassays, which use fluorescent substances to label antibodies or antigen molecules and detect changes in the intensity of the fluorescent signal after specific binding to the test sample, are more suitable for quantitative detection of allergen-specific IgE antibody levels. Quantum dots (QDS), a new type of fluorescent nanocrystal, offer high fluorescence efficiency, resistance to photobleaching, long fluorescence lifetime, narrow and symmetrical emission spectra, multiple emission per unit excitation, and the ability to detect multiple indicators simultaneously. Quantum dot microspheres in quantum dot microsphere markers encapsulate quantum dots within the microspheres, amplifying the quantum dot fluorescence signal. The coating not only stabilizes the quantum dots, improving their biocompatibility and colloidal stability, but also allows for the modification of various functional groups on the microsphere surface, enabling the binding of multiple markers to the microsphere surface. These properties of quantum dots provide an excellent option for preparing immunofluorescent probes, thereby enabling the quantitative detection of allergen-specific IgE antibodies in test samples using fluorescent immunoassays.

[0005] Currently, the allergen immunochromatographic quantitative detection products on the market are difficult to perform multiple tests simultaneously, and there are serious interferences during the detection of multiple test items. Therefore, there is an urgent need for an allergen immunochromatographic quantitative detection product that can ensure simultaneous multiple tests.

[0006] Summary of the Invention

[0007] The present application provides a multi-detection quantum dot quantitative detection card, preparation method, device and use method to solve the problem that allergen immunochromatographic quantitative detection products currently on the market are difficult to perform multiple detections at the same time and cause serious interference during the detection of multiple detection items.

[0008] In a first aspect, an embodiment of the present application provides a multi-detection quantum dot quantitative detection card, the detection card comprising:

[0009] Multiple cartridges and test strips;

[0010] A plurality of reaction slots are provided in the multi-row cartridge, and the reaction slots are used to place the test strips;

[0011] The multi-row cartridge further comprises a plurality of sample addition holes and a plurality of observation windows, wherein the sample addition holes, the observation windows and the reaction tanks correspond one to one;

[0012] The test strip includes a conjugate pad coated with a quantum dot-labeled antibody;

[0013] Wherein, the quantum dot-labeled antibody serves as a color-developing substance.

[0014] Optionally, the test strip further comprises a sample pad, a nitrocellulose membrane, absorbent paper and a bottom plate;

[0015] The sample pad, conjugate pad, nitrocellulose membrane and absorbent paper are uniformly pasted on the bottom plate in sequence;

[0016] Wherein, the nitrocellulose membrane is coated with multiple detection lines and a quality control line;

[0017] Wherein, the multiple detection lines and one quality control line are evenly spaced and fixed on the nitrocellulose membrane.

[0018] Optionally, the multiple detection lines are coated with different allergen antigens, and the quality control line is coated with sheep anti-chicken IgY antibody.

[0019] Optionally, the allergen antigen is at least any one of cat dander, mugwort, birch, timothy, cockroach, dog dander, plantain, ragweed, house dust mite, Alternaria (inhalation allergens), peanuts, milk, cod, scallops, egg white, shrimp, crab, soybeans, wheat, and almonds (ingestion allergens).

[0020] Optionally, the quantum dot-labeled antibodies coated on the conjugate pad are mouse anti-human IgE antibodies and chicken IgY antibodies.

[0021] A second aspect of the present invention provides a method for preparing a multi-detection quantum dot quantitative detection card, the method comprising the following steps:

[0022] Prepare a coated nitrocellulose membrane containing multiple test lines and quality control lines;

[0023] preparing a conjugate pad coated with two quantum dot-labeled antibodies, wherein the two quantum dot-labeled antibodies include: a quantum dot-labeled mouse anti-human IgE antibody and a quantum dot-labeled chicken IgY antibody;

[0024] The nitrocellulose membrane coated and the binding pad coated with the two quantum dot-labeled antibodies are assembled to obtain a multi-detection quantum dot quantitative detection card.

[0025] Optionally, the preparation of a coated nitrocellulose membrane comprising a plurality of detection lines and quality control lines comprises:

[0026] Fix the nitrocellulose membrane in the middle of the bottom plate;

[0027] The allergen antigen and antibody diluted to a desired coating concentration are fixed on a nitrocellulose membrane and dried to prepare the coated nitrocellulose membrane.

[0028] Optionally, the preparation of a conjugate pad coated with two quantum dot-labeled antibodies comprises:

[0029] Obtaining a quantum dot solution;

[0030] preparing quantum dot-labeled mouse anti-human IgE antibodies and quantum dot-labeled chicken IgY antibodies;

[0031] The quantum dot-labeled mouse anti-human IgE antibody and the quantum dot-labeled chicken IgY antibody are mixed in proportion and fixed on a conjugate pad, followed by drying to prepare the conjugate pad coated with the two quantum dot-labeled antibodies.

[0032] Optionally, assembling the nitrocellulose membrane-coated and the binding pad-coated with the two quantum dot-labeled antibodies comprises:

[0033] Assembling the coated nitrocellulose membrane, the binding pad coated with the two quantum dot-labeled antibodies, the sample pad, absorbent paper, and a bottom plate to obtain a test strip;

[0034] The test strip is fixedly placed in a placement slot in a multi-link card housing to obtain a multi-link quantum dot quantitative detection card.

[0035] Optionally, the nitrocellulose membrane, the conjugate pad and sample pad coated with the two quantum dot-labeled antibodies, absorbent paper, and a bottom plate are assembled to obtain a test strip, comprising:

[0036] Fixing absorbent paper on the edge of the nitrocellulose membrane coated near the quality control line, and fixing the binding pad coated with the two quantum dot-labeled antibodies on the edge of the nitrocellulose membrane coated near the detection line;

[0037] A sample pad is fixedly attached to the edge of the binding pad coated with the two quantum dot-labeled antibodies away from the detection line;

[0038] The bottom plate of the fixed adhesive coated nitrocellulose membrane, the absorbent paper, the binding pad coated with the two quantum dot labeled antibodies and the sample pad is cut into pieces of 2 to 6 mm to obtain the test strip.

[0039] The detection principle of the multi-test quantum dot quantitative test card is as follows: IgE antibodies in the test sample bind to quantum dot-labeled mouse anti-human IgE antibodies (chromogenic substances) and migrate along the test strip toward the test line and control line. On the test line, they bind to the coated allergen antigens to form a complex of "allergen antigen-sIgE-mouse anti-human IgE quantum dots." Exciting the quantum dots with an excitation light source produces a fluorescent signal. The higher the sIgE concentration, the higher the quantum dot content on the test line, and the stronger the excited fluorescent signal. By measuring the signal intensity, the IgE antibody content in the test sample can be inferred, achieving the purpose of quantitative testing. When quantum dot-labeled chicken IgY antibodies (chromogenic substances) migrate to the control line, a "sheep anti-chicken IgY-chicken IgY quantum dot" complex is formed, resulting in a signal. The absence of a fluorescent signal on the control line indicates a failure of the reaction system. The intensity of the fluorescent signal on the test line is proportional to the specific IgE antibody content in the sample. By comparing the fluorescent signal intensity to a standard curve, the specific IgE content in the sample can be determined.

[0040] This application has the following advantages:

[0041] 1) This application simultaneously uses quantum dot labeling for quantitative detection, multiple detection lines on a nitrocellulose membrane, and a multi-link cartridge format, solving both the quantitative and multi-link detection problems, thereby improving detection efficiency and accuracy.

[0042] 2) This application technically achieves the goal of rapidly diagnosing various allergic diseases caused by inhaled and ingested allergens in the human body;

[0043] 3) This application improves the chromatographic reaction effect of the test strips, while reducing the interference of simultaneous testing of multiple test strips. It is easy to operate, highly sensitive, and the quantitative test results can provide doctors with more accurate and effective assistance in clinical diagnosis.

[0044] 4) The blocking solution used in the process of quantum dot-labeled antibodies in this application is helpful in solving the crosstalk between detection items and non-specific interference in the detection samples.

[0045] The present application provides a multiple detection device and a method for using the same to solve the problem that colloidal gold chromatography products currently on the market are difficult to perform multiple detections with a single sample addition, and that multiple detection items have serious interference.

[0046] A third aspect of the embodiments of the present application provides a multi-link detection device, comprising: a detection card and a detection strip adapted to the detection card;

[0047] The test card comprises a first shell and a second shell, wherein the second shell is provided with two placement slots for placing the test strip;

[0048] The first shell is provided with a sample addition hole, and a diversion cavity is provided on the first shell at the position of the sample addition hole, a diversion part is provided in the diversion cavity, and the center of the diversion part is arranged opposite to the center of the sample addition hole;

[0049] The diversion cavity includes two liquid outlet holes, which are respectively located on both sides of the diversion portion, and the two liquid outlet holes are respectively communicated with two placement grooves.

[0050] Optionally, exhaust holes are provided on the side walls on both sides of the first shell, and positions of the two exhaust holes correspond to positions of the two liquid outlet holes respectively.

[0051] Optionally, two observation windows are provided on the first housing;

[0052] Wherein, the two observation windows are respectively arranged in one-to-one correspondence with the two placement slots.

[0053] Optionally, the detection strip includes a first end and a second end, and when the detection strip is located in the placement groove, the position of the first end corresponds to the position of the liquid outlet;

[0054] The test strip comprises a bottom plate and a sample pad, a colloidal gold pad, a nitrocellulose membrane and a blotting paper arranged on the bottom plate, wherein the sample pad, the colloidal gold pad, the nitrocellulose membrane and the blotting paper are arranged on the bottom plate in sequence from the first end to the second end;

[0055] Wherein, the nitrocellulose membrane is coated with multiple detection lines and quality control lines.

[0056] Optionally, the position of the nitrocellulose membrane corresponds to the position of the observation window.

[0057] Optionally, the plurality of test lines are coated with a plurality of allergen antigens, and the quality control line is coated with sheep anti-chicken IgY antibody;

[0058] Each of the test lines is coated with an allergen antigen, and the types of allergen antigens coated on each of the test lines are different.

[0059] Optionally, the plurality of allergen antigens include inhaled allergen antigens and ingested allergen antigens;

[0060] The inhaled allergen includes one of the following: cat dander, mugwort, birch, timothy grass, cockroach, dog dander, plantain, ragweed, house dust mite, and Alternaria;

[0061] The food allergens include one of the following: peanuts, milk, cod, scallops, egg white, shrimp, crab, soy, wheat and almonds.

[0062] Optionally, the antibodies coated on the colloidal gold pad are colloidal gold particles-mouse anti-human IgE antibodies and colloidal gold particles-chicken IgY antibodies.

[0063] A fourth aspect of the present application provides a method for using a multiplex detection device, wherein the detection device includes a detection card and a detection strip, and the detection card is provided with a sample addition well. The method includes:

[0064] Adding the test sample into the sample adding well;

[0065] Determine detection information generated by the detection strip according to the detection sample.

[0066] Optionally, the test sample is a whole blood sample, a plasma sample, or a serum sample.

[0067] Beneficial effects of this application:

[0068] 1) This application technically achieves the goal of rapidly detecting allergens (including inhaled or ingested allergens) that cause various allergic reactions in the human body. It can identify specific allergens, facilitating rapid testing by doctors and self-diagnosis by patients. Single-hole, single-time sample addition is convenient to operate, eliminating the need for multiple sample collections. The detection device is highly sensitive and portable, requiring no additional equipment for operation; it can be completed according to the instructions.

[0069] 2) This application provides a special test cartridge that can simultaneously test two test strips with a single sample loading, solving the pain point of traditional multi-test cartridges that require multiple loadings.

[0070] 3) This application is applicable to whole blood testing, especially for infants and young children and family self-testing. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments of the present application. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0072] FIG1 is a schematic structural diagram of a multi-detection quantum dot quantitative detection card provided in one embodiment of the present application;

[0073] FIG2 is a schematic diagram of a test strip structure of a multi-detection quantum dot quantitative detection card provided in one embodiment of the present application;

[0074] FIG3 is a graph showing the correlation test results of allergen-specific IgE antibody levels in serum samples according to an embodiment of the present application;

[0075] FIG4 is a schematic diagram of the structure of a detection card in a multi-detection device provided in one embodiment of the present application;

[0076] FIG5A is a cross-sectional view of the sample addition well in the AA direction in FIG4 ;

[0077] FIG5B is a partial enlarged schematic diagram of FIG5A;

[0078] FIG6A is a schematic diagram of the structure of the first housing of the detection card in FIG4 in the BB direction;

[0079] FIG6B is a partial enlarged schematic diagram of FIG6A;

[0080] FIG7 is a front perspective view of FIG4;

[0081] FIG8 is a schematic diagram of the structure of a test strip of a multi-detection device provided in one embodiment of the present application;

[0082] FIG9 is a flowchart of a method for using a multi-detection device according to an embodiment of the present application. Specific embodiments

[0083] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0084] Example 1

[0085] As shown in FIG1-2 , a multi-detection quantum dot quantitative detection card 100 and a detection strip 200 of this embodiment are shown.

[0086] As shown in FIG1 , a multi-detection quantum dot quantitative detection card 100 of this embodiment is shown.

[0087] The multi-detection quantum dot quantitative detection card 100 includes a multi-connection card housing 101 and a plurality of detection strips 200 .

[0088] A plurality of placement slots (not shown in the figure) are provided in the multi-row cartridge 101 , and the reaction slots are used to place the test strips 200 .

[0089] At the same time, the multi-row cartridge 101 further includes a plurality of sample addition holes 104 and a plurality of observation windows 103 . The sample addition holes are used for adding test samples, etc., and the observation windows are used for observing the specific conditions of the test strips.

[0090] Preferably, a buffer area (not marked in the figure) is further provided around the sample addition well 104. Since the flow of the test sample and the sample diluent requires a certain amount of time, the buffer area can be used to buffer the test sample after being added to the sample addition well.

[0091] The sample addition holes 104 , the observation windows 103 and the reaction tanks correspond one to one to facilitate the arrangement of the detection strips.

[0092] Preferably, the multi-row cartridge can be provided with three placement slots, three sample addition holes and three observation windows.

[0093] Preferably, the multi-detection quantum dot quantitative detection card is sealed in an aluminum foil bag containing a desiccant and stored.

[0094] As shown in FIG2 , a test strip 200 of a multi-test quantum dot quantitative test card of this embodiment is shown.

[0095] The test strip 200 includes a conjugate pad 213 coated with quantum dot-labeled antibodies.

[0096] Among them, quantum dot-labeled antibodies are used as color-developing substances.

[0097] Meanwhile, the test strip 200 further includes a sample pad 202, a nitrocellulose membrane 204, absorbent paper 207 and a base plate 201;

[0098] The sample pad 202 , the conjugate pad 213 , the nitrocellulose membrane 204 and the absorbent paper 207 are uniformly adhered to the bottom plate 201 in sequence.

[0099] Preferably, the sample pad 202 is a polyester fiber membrane or a glass cellulose membrane.

[0100] Preferably, the conjugate pad 213 is a glass cellulose membrane.

[0101] Preferably, the bottom plate 201 is a PVC board.

[0102] Preferably, the nitrocellulose membrane 204 is coated with multiple detection lines 206 and a quality control line 205. When in use, the detection lines are used to display the test results of the multi-detection quantum dot quantitative detection card, and the quality control line is used to display whether the test results of the multi-detection quantum dot quantitative detection card are valid.

[0103] Among them, the position of the sample pad corresponds to the position of the sample addition hole, so that the test sample can flow directly to the sample pad, reducing the loss of the test sample; the position of the nitrocellulose membrane corresponds to the position of the observation window, so as to detect the fluorescence signals of the test line and the quality control line.

[0104] Multiple test lines 206 and one quality control line 205 are evenly spaced and fixed on the nitrocellulose membrane. In the embodiment of the present application, there are three test lines in total, and the three test lines and one quality control line are evenly spaced 2 to 6 mm along the length of the test strip, for example, 2 mm, 3 mm, or 6 mm. The quality control line is located near the absorbent paper.

[0105] Three test lines are coated with different allergen antigens, and one quality control line is coated with goat anti-chicken IgY antibodies. The allergen antigens coated on the test lines bind to quantum dot-labeled antibodies (chromogenic substances) to form complexes. Laser irradiation of these complexes can detect corresponding fluorescent signals, which are then plugged into the standard curve to calculate the IgE antibody content in the test sample. The goat anti-chicken IgY antibodies coated on the quality control line bind to quantum dot-labeled antibodies to form complexes. The validity of the test results from the multi-test quantum dot quantitative test card can be determined by detecting whether the complexes produce signal values.

[0106] Preferably, the standard curve is prepared by configuring the international standard of total IgE antibody into the following concentrations: 0.1IU / mL, 0.3IU / mL, 1.2IU / mL, 4.8IU / mL, 19.2IU / mL, 38.3IU / mL, 76.6IU / mL, 100IU / mL, detecting the total IgE antibody at the above concentrations to obtain the signal value corresponding to each concentration: 0.0016, 0.0049, 0.0199, 0.0754, 0.2761, 0.5004, 0.8845, 1.1053, substituting the above data into the standard curve obtained by four-parameter fitting, the standard curve formula is y=(AD) / [1+(x / C)^B]+D, A=9.67167, B=-0.89759, C=980.14709, D=-0.00171, R 2 =0.99995.

[0107] Preferably, the international standard for total IgE antibodies is human serum immunoglobulin E (IgE) international standard, NIBSC code: 11 / 234.

[0108] Preferably, each test line is coated with one allergen antigen, and the allergen antigens coated on each test line are different.

[0109] Preferably, the multiple allergen antigens are any one of cat dander, mugwort, birch, timothy, cockroach, dog dander, plantain, ragweed, house dust mite, Alternaria alternifolia (inhalation allergens), peanuts, milk, cod, scallops, egg white, shrimp, crab, soybeans, wheat, and almonds (ingestion allergens).

[0110] For example, the plurality of detection lines include a first detection line, a second detection line, and a third detection line, wherein the first detection line is coated with a crude peanut antigen extract, the second detection line is coated with a crude cockroach antigen extract, and the third detection line is coated with a crude soybean antigen extract. Those skilled in the art can configure these lines as needed, and this embodiment of the present application will not be described in detail.

[0111] Preferably, the quantum dot-labeled antibodies coated on the conjugate pad are mouse anti-human IgE antibodies and chicken IgY antibodies.

[0112] The test results of the multi-detection quantum dot quantitative detection card for the simultaneous detection of multiple different inhaled and ingested allergens are shown in Table 1. The results show that the multi-detection quantum dot quantitative detection card can simultaneously detect the content of IgE antibodies to different allergens in the test sample, and after ten repeated tests, the test results of the ten tests are highly precise. The high test precision ensures the accuracy of the test results of the multi-detection quantum dot quantitative detection card. Therefore, the present application simultaneously adopts quantum dot labeling quantitative detection, multiple detection lines are set on the nitrocellulose membrane, and the form of a multi-linked card shell, which solves the problems of quantitative detection and multi-link detection at the same time, and improves the detection efficiency and detection accuracy of the multi-detection quantum dot quantitative detection card.

[0113] Table 1 Results of allergen-specific IgE antibody content in serum samples (IU / mL)

[0114] Because the result of the Fadia test kit is the gold standard in the detection of allergen-specific IgE antibodies, the Fadia test kit and the multi-detection quantum dot quantitative detection reagent card are used to simultaneously detect samples containing specific allergen IgE antibodies of at least any one of the inhaled allergens and ingested allergens described in this application. The relevant test results are shown in Table 2 and Figure 3 (the test result A is the test result of the Fadia test kit, and the test result B is the test result of the multi-detection quantum dot quantitative detection card). According to Figure 3, the correlation formula of the test results of the Fadia test kit and the multi-detection quantum dot quantitative detection reagent card is y=0.9244x+0.2388, R 2 =0.957. Figure 3 shows that the test results of the present application are well correlated with the test results of the Fadia test kit, which is recognized as the gold standard in the industry. It can be seen that the test results of the multi-detection quantum dot quantitative detection card in the present application are more accurate.

[0115] Table 2 Correlation test results of allergen-specific IgE antibody content in serum samples (IU / mL)

[0116] Example 2

[0117] This embodiment discloses a method for preparing a multi-detection quantum dot quantitative detection card, which comprises the following steps:

[0118] S1, prepare a coated nitrocellulose membrane containing multiple test lines and quality control lines.

[0119] Specifically, preparing a coated nitrocellulose membrane containing multiple test lines and quality control lines includes:

[0120] Fix the nitrocellulose membrane in the middle of the bottom plate;

[0121] The allergen antigen and antibody diluted to a desired coating concentration are fixed on a nitrocellulose membrane and dried to prepare the coated nitrocellulose membrane.

[0122] Among them, the allergen antigen is any one of cat dander, mugwort, birch, timothy, cockroach, dog dander, plantain, ragweed, house dust mite, Alternaria mold (inhalation allergens), peanuts, milk, cod, scallops, egg white, shrimp, crab, soybeans, wheat, and almonds (ingestion allergens); the antibody is sheep anti-chicken IgY antibody.

[0123] Preferably, the desired coating concentration of the allergen antigen is 0.5-2 mg / mL. For example, the concentration of the coated allergen antigen can be 0.5 mg / mL, 1 mg / mL, 1.5 mg / mL, 2 mg / mL, etc. Those skilled in the art can adjust the concentration as needed, and this embodiment will not be described in detail.

[0124] Preferably, the required concentration of the coated goat anti-chicken IgY antibody is 1 to 3 mg / mL. For example, the concentration of the coated goat anti-chicken IgY antibody can be 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, etc. Those skilled in the art can set this concentration as needed, and this embodiment of the present application will not be described in detail.

[0125] Preferably, the nitrocellulose membrane coated with the allergen antigen and antibody is dried at 37°C overnight.

[0126] S2, preparing a binding pad coated with two quantum dot-labeled antibodies, wherein the two quantum dot-labeled antibodies include: a quantum dot-labeled mouse anti-human IgE antibody and a quantum dot-labeled chicken IgY antibody.

[0127] Specifically, preparing a conjugate pad coated with two quantum dot-labeled antibodies comprises:

[0128] Obtaining a quantum dot solution;

[0129] The quantum dots were added to a pH = 6 4-morpholineethanesulfonic acid buffer, and a certain mass ratio of EDC (1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydro, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide) and NHS (NHS N-Hydroxysuccinimide, N-hydroxysuccinimide) were added in sequence. After rotating and mixing for 30 minutes, the mixture was activated and the supernatant was removed by centrifugation. The quantum dot solid was added to a pH = 6 4-morpholineethanesulfonic acid buffer or a pH = 8 borate buffer and resuspended by shaking to obtain a quantum dot solution.

[0130] Preferably, the mass ratio of the added EDC and NHS is 1:2.

[0131] preparing quantum dot-labeled mouse anti-human IgE antibodies and quantum dot-labeled chicken IgY antibodies;

[0132] Mouse anti-human IgE antibody or chicken IgY antibody was added to the above quantum dot solution, placed on a rotating mixer, and rotated for 2 hours. After coupling, blocking solution was added, and after rotating for 30 minutes, the supernatant was removed and the quantum dot solid was resuspended with a binding pad treatment solution to obtain quantum dot-labeled mouse anti-human IgE antibody or quantum dot-labeled chicken IgY antibody.

[0133] Preferably, the concentration of the quantum dot-labeled mouse anti-human IgE antibody or the quantum dot-labeled chicken IgY antibody is 20 to 300 μg / mL. For example, the concentration of the quantum dot-labeled mouse anti-human IgE antibody or the quantum dot-labeled chicken IgY antibody can be 20 μg / mL, 70 μg / mL, 110 μg / mL, 150 μg / mL, 180 μg / mL, 220 μg / mL, 260 μg / mL, 300 μg / mL, etc. Those skilled in the art can set it according to needs, and the embodiments of this application will not be repeated in detail.

[0134] Preferably, the blocking solution contains 1-10% casein, 1-10% PVP-30

[0135] A mixed solution of at least one of (Polyvinylpyrrolidone K30), 1-10% mouse IgG antibody, and 1-10% glycine is used as a blocking solution to help solve the problems of cross-detection items and non-specific interference in detection samples.

[0136] The conjugate pad treatment solution contains sugars, macromolecular polymers, surfactants, diluents, proteins, and preservatives.

[0137] Preferably, the sugar is at least one of trehalose, sucrose and dextran.

[0138] Preferably, the macromolecular polymer is at least one of PEG-20000 (Polyethylene glycol-20000), PVP-30, PVP-10 (Polyvinylpyrrolidone K10), and PEG-8000 (Polyethylene glycol-8000).

[0139] Preferably, the surfactant is at least one of TX-100 (Triton X-100), TW-20 (Tween-20), and S9 (Pluracare 1307, propylene oxide-ethylene oxide-ethylene diamine copolymer).

[0140] Preferably, the diluent is at least one of PBS buffer (phosphate buffered saline), Tris (Tris (hydroxymethyl) aminomethane) buffer, and BBS buffer (Borate Buffered Salin).

[0141] Preferably, the protein is at least one of casein, bovine serum albumin, and skim milk.

[0142] Preferably, the preservative is at least one of NaN3 (Sodium azide) and PC300 (ProClin300).

[0143] Among them, macromolecular polymers are beneficial to improving antibody activity, surfactants are beneficial to liquid chromatography reactions, proteins are beneficial to liquid chromatography and reducing non-specific binding in samples, and preservatives are beneficial to improving the stability of the binding pad.

[0144] The quantum dot-labeled mouse anti-human IgE antibody and the quantum dot-labeled chicken IgY antibody are mixed in proportion and fixed on a conjugate pad, and then dried to prepare a conjugate pad coated with the two quantum dot-labeled antibodies.

[0145] Preferably, the quantum dot-labeled mouse anti-human IgE antibody and the quantum dot-labeled chicken IgY antibody are mixed in a ratio of 3:1 to 10:1. For example, the ratio of the quantum dot-labeled mouse anti-human IgE antibody to the quantum dot-labeled chicken IgY antibody can be 3:1, 5:1, 8:1, 10:1, etc. Those skilled in the art can configure this as needed, and the present embodiment will not be described in detail.

[0146] Preferably, the conjugate pad coated with the two quantum dot-labeled antibodies is dried at 37° C. overnight.

[0147] S3, assembling the nitrocellulose membrane coated and the binding pad coated with the two quantum dot-labeled antibodies to obtain a multi-detection quantum dot quantitative detection card.

[0148] Specifically, assembling a nitrocellulose membrane-coated binding pad and two quantum dot-labeled antibodies-coated binding pads includes:

[0149] Assembling the coated nitrocellulose membrane, the binding pad coated with the two quantum dot-labeled antibodies, the sample pad, absorbent paper, and a bottom plate to obtain a test strip;

[0150] Particularly, absorbent paper is fixedly pasted on the edge of the nitrocellulose membrane coated near the quality control line, and the binding pad coated with the two quantum dot-labeled antibodies is fixedly pasted on the edge of the nitrocellulose membrane coated near the detection line; the sample pad is fixedly pasted on the edge of the binding pad coated with the two quantum dot-labeled antibodies away from the detection line; and the bottom plate to which the nitrocellulose membrane coated, the absorbent paper, the binding pad coated with the two quantum dot-labeled antibodies, and the sample pad are fixedly pasted is cut into 3 mm to obtain the test strip.

[0151] The test strip is fixedly placed in a placement slot in a multi-link card housing to obtain a multi-link quantum dot quantitative detection card.

[0152] Example 3

[0153] As shown in FIG. 4 to FIG. 8 , a multiple detection device according to this embodiment includes a detection card 100 and a detection strip 200 adapted to the detection card.

[0154] As shown in Figures 4 to 7, a detection card of a multi-link detection device of this embodiment is shown.

[0155] The test card 100 includes a first shell 101 and a second shell 102 (not shown in the figure). The first shell 101 is provided with a sample addition hole 104 for adding a test sample at a time.

[0156] A diversion cavity is provided on the first housing 101 at the location of the sample addition hole 104 . A diversion portion 105 is provided in the diversion cavity. The diversion portion 105 is used to divert the test sample added to the sample addition hole 104 into two parts.

[0157] The center of the diversion part 105 is arranged opposite to the center of the sample addition hole 104. The diversion cavity includes two liquid outlet holes 106, which are respectively located on both sides of the diversion part 105. After the diversion part 105 diverts the test sample into two parts, the two parts of the test sample can flow out from the two liquid outlet holes 106 respectively.

[0158] At the same time, two placement slots (not marked in the figure) are provided on the second shell 102, which are used to place the test strip 100; when the first shell 101 and the second shell 102 are connected, the two liquid outlets 106 are respectively connected to the two placement slots, so that the test samples after diversion can directly enter the placement slots and be directly used for test strip detection, avoiding the loss of test samples and reducing the amount of test samples added.

[0159] The first housing 101 is also provided with a buffer area (not marked in the figure) for the sample addition hole 104. Since the flow of the test sample and the sample diluent requires a certain amount of time, the buffer area can be used to buffer the test sample and the sample diluent after being added to the sample addition hole.

[0160] Preferably, the buffer area and the sample addition hole 104 form a frustum structure, and the sample addition hole 104 is a circle with a smaller diameter in the frustum structure; the diameter of the sample addition hole is 11.6 mm, and the diameter of the liquid outlet hole is 2.46 mm.

[0161] Preferably, the diverter is a wedge-shaped component including two inclined surfaces, and the inclination directions of the two inclined surfaces are symmetrical about the center of the diverter, and the angle between the inclined surfaces and the vertical direction is preferably 110°.

[0162] Preferably, the diversion portion is made of one of ABS (Acrylonitrile Butadiene Styrene) plastic, PS (polystyrene) plastic, HIPS (High impact polystyrene) plastic, and PC (Polycarbonate) plastic.

[0163] Since the test sample is a blood sample, including a whole blood sample, a plasma sample, or a serum sample, the relative viscosity of the blood sample is greater than that of water, resulting in the blood sample not easily flowing downward when added to the sample addition hole. Therefore, in order to improve the fluidity of the test sample, in the embodiment of the present application, exhaust holes 107 are provided on the side walls on both sides of the first shell 101, and the positions of the two exhaust holes 107 correspond to the positions of the two liquid outlet holes 106, respectively. Therefore, after the test sample is added, the air in the liquid outlet hole 106 is discharged through the exhaust holes 107, allowing the test sample to flow smoothly without blocking the sample addition hole 104.

[0164] At the same time, two observation windows 103 are provided on the first shell 101. The two observation windows 103 are respectively provided in one-to-one correspondence with the two placement slots, so that the user can observe the specific conditions of the test strips.

[0165] After the test card 100 is assembled, it is sealed and placed in an aluminum foil bag containing a desiccant for storage.

[0166] FIG8 shows a test strip in a multi-detection device according to the present embodiment.

[0167] The test strip 200 includes a first end and a second end. When in use, the test strip 200 is located in the placement groove, and the position of the first end corresponds to the position of the liquid outlet 106 .

[0168] The test strip 200 includes a base plate 201 and a sample pad 202, a colloidal gold pad 203, a nitrocellulose membrane 204, and a blotting paper 207 disposed on the base plate. The sample pad 202, the colloidal gold pad 203, the nitrocellulose membrane 204, and the blotting paper 207 are sequentially disposed on the base plate 201 from a first end to a second end. That is, the sample pad 202 is disposed corresponding to the liquid outlet 106, so that the test sample flowing out of the liquid outlet 106 can be absorbed by the sample pad 202, thereby reducing the loss of the test sample.

[0169] It should be noted that the first end and the second end involved in this application are defined according to the flow direction of the test sample. During the use of the test strip, the test sample flows from the sample end to the absorbent paper through chromatography. The sample pad end is at the position of the first end, and the absorbent paper end is at the position of the second end.

[0170] Preferably, the sample pad 202 is a polyester fiber membrane or a glass cellulose membrane.

[0171] Preferably, the colloidal gold pad 203 is a glass cellulose membrane.

[0172] Preferably, the nitrocellulose membrane 204 is coated with a plurality of test lines 206 and a quality control line 205. During use, the test lines are used to display the test results of the multiple detection device, and the quality control line is used to indicate whether the test results of the multiple detection device are valid. In the embodiment of the present application, a total of five test lines are provided, and the five test lines are evenly spaced along the length of the test strip near the first end, and the quality control line is provided near the second end.

[0173] The position of the nitrocellulose membrane corresponds to the position of the observation window, so that the user can observe the detection results of the detection device.

[0174] Five test lines are coated with various allergen antigens, while the control line is coated with goat anti-chicken IgY antibodies. The allergen antigens on the test lines bind to colloidal gold particles coupled with mouse anti-human IgE antibodies, producing a color that can be observed visually. The control line, coated with goat anti-chicken IgY antibodies, binds to colloidal gold particles coupled with mouse anti-human IgE antibodies and colloidal gold particles coupled with chicken IgY antibodies, respectively, resulting in a red color, ensuring proper detection of the test strip.

[0175] Each test line is coated with an allergen antigen, and the type of allergen antigen coated on each test line is different.

[0176] Preferably, the plurality of allergen antigens include inhaled allergen antigens and ingested allergen antigens.

[0177] Preferably, the inhalant allergen comprises one of the following: cat dander, mugwort, birch, timothy, cockroach, dog dander, plantain, ragweed, house dust mite, and Alternaria.

[0178] Preferably, the ingested allergen includes one of the following: peanuts, milk, cod, scallops, egg white, shrimp, crab, soy, wheat and almonds.

[0179] For example, the plurality of test lines include a first test line, a second test line, a third test line, a fourth test line, and a fifth test line, wherein the first test line is coated with a crude antigen extract of cat dander, the second test line is coated with a crude antigen extract of peanuts, the third test line is coated with a crude antigen extract of birch, the fourth test line is coated with a crude antigen extract of egg white, and the fifth test line is coated with a crude antigen extract of cockroaches. Those skilled in the art can configure these as needed, and the embodiments of this application will not be described in detail.

[0180] Preferably, the concentration of the coated allergen antigen is 0.1-3 mg / mL. For example, the concentration of the coated allergen antigen can be 0.1 mg / mL, 0.2 mg / mL, 0.5 mg / mL, 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, etc. Those skilled in the art can adjust the concentration as needed, and this embodiment will not be described in detail.

[0181] Preferably, the required concentration of the coated goat anti-chicken IgY antibody is 1 to 3 mg / mL. For example, the concentration of the coated goat anti-chicken IgY antibody can be 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, etc. Those skilled in the art can set this concentration as needed, and this embodiment of the present application will not be described in detail.

[0182] Preferably, the antibodies coated on the colloidal gold pad are colloidal gold particles-mouse anti-human IgE antibody and colloidal gold particles-chicken IgY antibody.

[0183] Preferably, the colloidal gold particles-mouse anti-human IgE antibody and the colloidal gold particles-chicken IgY antibody coated with the colloidal gold pad are mixed in a ratio of 10:1 to 3:1. For example, the ratio of colloidal gold particles-mouse anti-human IgE antibody to colloidal gold particles-chicken IgY antibody can be 10:1, 8:1, 5:1, 3:1, etc. Those skilled in the art can set it according to their needs, and the embodiments of this application will not be repeated in detail.

[0184] Preferably, the concentration of the colloidal gold particles-mouse anti-human IgE antibody and the colloidal gold particles-chicken IgY antibody coated on the colloidal gold pad is 5 to 25 μg / mL. For example, the concentration of the colloidal gold particles-mouse anti-human IgE antibody and the colloidal gold particles-chicken IgY antibody can be 5 μg / mL, 10 μg / mL, 15 μg / mL, 25 μg / mL, etc. Those skilled in the art can set it according to their needs, and the embodiments of this application will not be repeated in detail.

[0185] The specific method for preparing the test strip may include: attaching the coated nitrocellulose membrane 204 to the middle of the base plate 201; attaching one end of the absorbent paper 207 to the second end of the base plate, and covering the other end of the absorbent paper 207 on the nitrocellulose membrane 204; covering the nitrocellulose membrane 204 near the detection line 206 at one end of the colloidal gold pad 203, and attaching the other end to the base plate 201; attaching one end of the sample pad 202 to the first end of the base plate 201, and covering the other end of the colloidal gold pad 203; cutting it into 3 mm wide strips with a cutting machine, and then obtaining a test strip 200 of a multi-detection device of the present application.

[0186] This embodiment also discloses a multiplex test device that includes, in addition to the test card and test strips, a sample diluent. During use, the sample diluent must be used in conjunction with the test sample. After the test sample is added, the sample diluent is added. The sample diluent helps prevent the test sample from coagulating too quickly, thus avoiding test failure. It also helps dilute the concentration of the test sample, improving test accuracy.

[0187] The sample diluent comprises a buffer, a macromolecular polymer, a surfactant, and a preservative. These components help stabilize the spatial structure of the antibody and improve the chromatography of the blood sample on the nitrocellulose membrane.

[0188] Preferably, the buffer is at least one of PBS (phosphate buffered saline), Tris (Tris(hydroxymethyl)aminomethane), and BBS (borate buffered saline); the macromolecular polymer is at least one of PEG (polyethylene glycol), PVP (polyvinyl pyrrolidone), and PVA (polyvinyl alcohol); the surfactant is at least one of Triton X-ray and Tween; and the preservative is at least one of NaN3 (sodium azide) and PC-300 (ProClin300). This embodiment also discloses the components of the sample pad treatment solution in the test strip.

[0189] The sample pad treatment fluid consists of a buffer, a macromolecular polymer, a surfactant, a preservative, a blocking agent, and substances that facilitate whole blood sample processing. When used with whole blood samples, these components help filter out large particles, enhance specific reactions, reduce interference from endogenous substances in the sample, and improve the sample's performance on nitrocellulose membrane chromatography.

[0190] Preferably, the blocking agent is at least one of a human anti-animal antibody (such as a human anti-mouse antibody) blocker, a rheumatoid factor blocker, a heterophile antibody blocker, a complement blocker, and an autoantibody blocker; and the substance that is beneficial to the processing of whole blood samples is at least one of an anti-erythrocyte antibody and a phytohemagglutinin. This embodiment also discloses the components of the colloidal gold pad treatment solution in the test strip, including a buffer, a macromolecular polymer, a surfactant, a preservative, and a carbohydrate. The components added to the colloidal gold pad treatment solution help to improve the sensitivity of the antibody and the stability of the antibody in the solid phase.

[0191] Preferably, the sugar is at least one of trehalose, sucrose and dextran.

[0192] Preferably, the buffer concentration in the sample diluent, sample pad treatment liquid and colloidal gold pad treatment liquid is 10-200 mM, the macromolecular polymer concentration is 0.1%-5%, the surfactant concentration is 0.1%-3%, the preservative concentration is 0.01%-2%, the concentration of the substance that is conducive to whole blood sample processing is 10-50 μg / mL, and the carbohydrate concentration is 0.5%-5%.

[0193] After the sample pad and colloidal gold pad on the test strips in the test device were treated with the aforementioned sample pad treatment fluid and colloidal gold pad treatment fluid optimization, the test device's positive and negative allergen detection results were optimized. The test devices (a multi-link test device without pad treatment fluid optimization and a multi-link test device with pad treatment fluid optimization) were coated with different inhaled allergens and the test results are shown in Tables 3 and 4. The results show that the positive and negative detection rates of the test devices meet industry standards, and the positive and negative detection rates of the optimized test devices are higher than those of the unoptimized test devices.

[0194] Table 3. Serum sample test results (1)

[0195] Table 4. Serum sample test results (2)

[0196] The macromolecular polymers added to the sample diluent, sample pad treatment solution, and colloidal gold pad treatment solution can enhance antibody activity and provide a spatial framework for the antibodies. Blocking agents help enhance specific reactions, reduce interference from endogenous substances in the sample, and mitigate interference caused by non-target protein antibodies in the test sample. Substances that facilitate whole blood sample processing can filter out large particles in the test sample, allowing the test sample to pass smoothly through the nitrocellulose membrane. Preservatives help stabilize the sample pad and colloidal gold pad. Therefore, test samples treated with the sample diluent, combined with optimized test strips, can improve the positive and negative coincidence rates of the test device.

[0197] Example 4

[0198] FIG9 shows a flowchart of steps of a method for using a multi-detection device.

[0199] 9 , this embodiment further discloses a method for using the multi-detection device, the method comprising:

[0200] Step 601: Add the test sample into the sample well.

[0201] Specifically, the test sample is a blood sample, a whole blood sample, a plasma sample, or a serum sample.

[0202] Specifically, the detectable allergens include: cat dander, mugwort, birch, timothy, cockroach, dog dander, plantain, ragweed, house dust mite, Alternaria, peanuts, milk, cod, scallops, egg white, shrimp, crab, soybeans, wheat, and almonds.

[0203] Step 602: Determine the test information generated by the test strip based on the test sample.

[0204] Specifically, 60 μL of the test sample is added dropwise from the sample addition hole, and then 60 μL of the sample diluent is added dropwise. The test sample and the sample diluent flow through the diversion part to the two liquid outlet holes and flow to the sample pads of the two test strips in the test card.

[0205] Furthermore, the diversion part divides the test sample and the sample diluent into two parts, and each part of the test sample and the sample diluent flows toward the liquid outlet through the two inclined surfaces of the diversion part.

[0206] Preferably, the two test strips disposed in the second housing are separated from each other; each test sample and sample diluent can enable the two test strips to undergo immunochromatographic reactions simultaneously.

[0207] Preferably, the detection device can simultaneously complete the detection of ten different allergens.

[0208] The detection device in this application only needs to perform a single-hole single-time sample addition, without the need to collect samples from the tester multiple times and multiple sample additions, and can detect ten different allergens at one time. It is easy to operate and highly practical.

[0209] The present application adopts highly specific IgE antibody and antigen reaction, and colloidal gold particles-mouse anti-human IgE antibodies and colloidal gold particles-chicken IgY antibodies are fixed on the colloidal gold pad. At the same time, combined with immunochromatography technology, after the test sample is diluted by the sample diluent, it combines with the colloidal gold particles-mouse anti-human IgE antibodies coated on the colloidal gold pad to form a marker during the flow process, and combines with the antigen coated on the test line to form red or pink spots visible to the naked eye. Finally, the sheep anti-chicken IgY antibodies coated on the quality control line will combine with the remaining colloidal gold particles-mouse anti-human IgE antibodies and colloidal gold particles-chicken IgY antibodies to form red or pink spots visible to the naked eye. Because the antibodies labeled with colloidal gold particles gather in large quantities at the corresponding ligands, they eventually develop color at the allergen antigen coated on the test line on the nitrocellulose membrane, and the test personnel can observe the test results of the multi-detection device with the naked eye through the observation window.

[0210] The detection strip of the multi-detection device in the present application is coated with multiple detection lines. After the detection sample is combined with the colloidal gold particles-mouse anti-human IgE antibodies, it is combined with the allergen antigens in the multiple detection lines. The following situations may occur: 1) The remaining colloidal gold particles-mouse anti-human IgE antibodies flowing to the quality control line are relatively high, causing the quality control line to show color; 2) The colloidal gold particles-mouse anti-human IgE antibodies flowing to the quality control line are too low or absent, causing the quality control line to not show color. Therefore, in order to avoid the occurrence of the second situation mentioned above, the colloidal gold pad is coated with colloidal gold particles-chicken IgY antibodies in addition to being coated with colloidal gold particles-mouse anti-human IgE antibodies. When the second situation mentioned above occurs, the sheep anti-chicken IgY antibodies coated on the quality control line can still combine with the colloidal gold particles-chicken IgY antibodies to show color, ensuring that the test results of the detection device are valid (negative or positive). Therefore, the multi-detection device prepared in the present application can not only perform specific detection of multiple allergen antigens at the same time, but also avoid the invalidation of the test strips in the detection device and reduce the need for repeated testing.

[0211] The analysis of the test strip test results is as follows:

[0212] Positive: When a red stripe appears on the control line and the test line also appears on the test line, it is judged as positive, indicating that the test result is that the IgE antibody corresponding to the test line in the test sample is positive;

[0213] Negative: When a red stripe appears on the quality control line and none of the test lines show red, it is judged as negative, indicating that the test result is that the IgE antibody corresponding to the test line in the test sample is negative;

[0214] Invalid: When the quality control line does not show red, regardless of whether a red stripe appears on the test line, it means that the test result is invalid.

[0215] The above is a detailed introduction to a multi-detection device and its use method provided by the present application. Specific examples are used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the method and core idea of ​​the present application. At the same time, for those skilled in the art, according to the ideas of the present application, there will be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as a limitation on the present application.

[0216] It should be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like used above to indicate orientations or positional relationships are based on the orientations or positional relationships shown in the accompanying drawings, and are only used to facilitate the description of this application and simplify the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on this application. In addition, the terms "first" and "second" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of this application, unless otherwise specified, the meaning of "multiple" is two or more.

[0217] The above description is merely a preferred embodiment of the present application and an illustration of the technical principles employed. Those skilled in the art should understand that the scope of the invention involved in this application is not limited to the technical solutions formed by the specific combination of the above-mentioned technical features, but also encompasses other technical solutions formed by any combination of the above-mentioned technical features or their equivalents without departing from the inventive concept. For example, a technical solution formed by replacing the above-mentioned features with (but not limited to) technical features with similar functions disclosed in this application.

Claims

1. A multi-detection quantum dot quantitative detection card, wherein: The test card comprises: Multiple cartridges and test strips; A plurality of reaction slots are provided in the multi-row cartridge, and the reaction slots are used to place the test strips; The multi-row cartridge further comprises a plurality of sample addition holes and a plurality of observation windows, wherein the sample addition holes, the observation windows and the reaction tanks correspond one to one; The test strip includes a conjugate pad coated with a quantum dot-labeled antibody; Wherein, the quantum dot-labeled antibody serves as a color-developing substance.

2. The multi-detection quantum dot quantitative detection card according to claim 1, wherein: The test strip also includes a sample pad, a nitrocellulose membrane, absorbent paper and a base plate; The sample pad, conjugate pad, nitrocellulose membrane and absorbent paper are evenly pasted on the bottom plate in sequence; The nitrocellulose membrane is coated with a plurality of detection lines and a quality control line, and the plurality of detection lines and the quality control line are evenly spaced and fixed on the nitrocellulose membrane.

3. The multi-detection quantum dot quantitative detection card according to claim 2, wherein: The multiple detection lines are coated with different allergen antigens, and the quality control line is coated with sheep anti-chicken IgY antibody.

4. The multi-detection quantum dot quantitative detection card according to claim 3, wherein: The allergen antigen is at least any one of cat dander, mugwort, birch, timothy, cockroach, dog dander, plantain, ragweed, house dust mite, Alternaria alternifolia (inhalation allergens), peanuts, milk, cod, scallops, egg white, shrimp, crab, soybean, wheat, and almond (ingestion allergens).

5. The multi-detection quantum dot quantitative detection card according to claim 1, wherein: The quantum dot-labeled antibodies coated on the conjugate pad are mouse anti-human IgE antibody and chicken IgY antibody.

6. A method for preparing a multi-detection quantum dot quantitative detection card, wherein: For preparing the multi-detection quantum dot quantitative detection card according to any one of claims 1 to 5, the preparation method comprises the following steps: Prepare a coated nitrocellulose membrane containing multiple test lines and quality control lines; preparing a conjugate pad coated with two quantum dot-labeled antibodies, wherein the two quantum dot-labeled antibodies include: a quantum dot-labeled mouse anti-human IgE antibody and a quantum dot-labeled chicken IgY antibody; The binding pad coated with nitrocellulose membrane and the binding pad coated with two quantum dot labeled antibodies Assemble and obtain a multi-detection quantum dot quantitative detection card.

7. The method for preparing the multi-detection quantum dot quantitative detection card according to claim 6, wherein: The method of preparing a coated nitrocellulose membrane comprising a plurality of test lines and quality control lines comprises: Fix the nitrocellulose membrane in the middle of the bottom plate; The allergen antigen and antibody diluted to a desired coating concentration are fixed on a nitrocellulose membrane and dried to prepare the coated nitrocellulose membrane.

8. The method for preparing the multi-detection quantum dot quantitative detection card according to claim 6, wherein: The method of preparing a conjugate pad coated with two quantum dot-labeled antibodies comprises: Obtaining a quantum dot solution; preparing quantum dot-labeled mouse anti-human IgE antibodies and quantum dot-labeled chicken IgY antibodies; The quantum dot-labeled mouse anti-human IgE antibody and the quantum dot-labeled chicken IgY antibody are mixed in proportion and fixed on a conjugate pad, followed by drying to prepare the conjugate pad coated with the two quantum dot-labeled antibodies.

9. The method for preparing the multi-detection quantum dot quantitative detection card according to claim 6, wherein: Assembling the nitrocellulose membrane and the binding pad coated with the two quantum dot-labeled antibodies comprises: Assembling the coated nitrocellulose membrane, the binding pad coated with the two quantum dot-labeled antibodies, the sample pad, absorbent paper, and a bottom plate to obtain a test strip; The test strip is fixedly placed in a placement slot in a multi-link card housing to obtain a multi-link quantum dot quantitative detection card.

10. The method for preparing the multi-detection quantum dot quantitative detection card according to claim 9, wherein: The nitrocellulose membrane, the binding pad and sample pad coated with two quantum dot-labeled antibodies, absorbent paper, and a bottom plate are assembled to obtain a test strip, comprising: Fixing absorbent paper on the edge of the nitrocellulose membrane coated near the quality control line, and fixing the binding pad coated with the two quantum dot-labeled antibodies on the edge of the nitrocellulose membrane coated near the detection line; A sample pad is fixedly attached to the edge of the binding pad coated with the two quantum dot-labeled antibodies away from the detection line; The bottom plate of the fixed adhesive coated nitrocellulose membrane, the absorbent paper, the binding pad coated with the two quantum dot labeled antibodies and the sample pad is cut into pieces of 2 to 6 mm to obtain the test strip.

11. A multi-link detection device, wherein: include: A test card and a test strip compatible with the test card; The test card comprises a first shell and a second shell, wherein the second shell is provided with two placement slots for placing the test strip; The first shell is provided with a sample addition hole, and a diversion cavity is provided on the first shell at the position of the sample addition hole, a diversion part is provided in the diversion cavity, and the center of the diversion part is arranged opposite to the center of the sample addition hole; The diversion cavity includes two liquid outlet holes, which are respectively located on both sides of the diversion portion, and the two liquid outlet holes are respectively communicated with two placement grooves.

12. The multi-detection device according to claim 11, wherein: Exhaust holes are provided on the side walls on both sides of the first shell, and the positions of the two exhaust holes correspond to the positions of the two liquid outlet holes respectively.

13. The multi-link detection device according to claim 11, wherein: Two observation windows are provided on the first shell; The two observation windows are respectively arranged in one-to-one correspondence with the two placement slots.

14. The multi-link detection device according to claim 11, wherein: The detection strip comprises a first end and a second end, and when the detection strip is located in the placement groove, the position of the first end corresponds to the position of the liquid outlet; The test strip comprises a bottom plate and a sample pad, a colloidal gold pad, a nitrocellulose membrane and a blotting paper arranged on the bottom plate, wherein the sample pad, the colloidal gold pad, the nitrocellulose membrane and the blotting paper are arranged on the bottom plate in sequence from the first end to the second end; Wherein, the nitrocellulose membrane is coated with multiple detection lines and quality control lines.

15. The multi-link detection device according to claim 14, wherein: The position of the nitrocellulose membrane corresponds to the position of the observation window.

16. The multi-link detection device according to claim 14, wherein: The plurality of test lines are coated with a variety of allergen antigens, and the quality control line is coated with sheep anti-chicken IgY antibody; Each of the test lines is coated with an allergen antigen, and the types of allergen antigens coated on each of the test lines are different.

17. The multi-link detection device according to claim 16, wherein: The multiple allergen antigens include inhaled allergen antigens and ingested allergen antigens; The inhaled allergen includes one of the following: cat dander, mugwort, birch, timothy grass, cockroach, dog dander, plantain, ragweed, house dust mite, and Alternaria; The food allergens include one of the following: peanuts, milk, cod, scallops, egg white, shrimp, crab, soy, wheat and almonds.

18. The multi-link detection device according to claim 14, wherein: The antibodies coated on the colloidal gold pad are colloidal gold particles-mouse anti-human IgE antibodies and colloidal gold particles-chicken IgY antibodies.

19. A method for using a multiplex detection device, the detection device comprising a detection card and a detection strip, the detection card being provided with a sample addition well, wherein: The method of use includes: Adding the test sample into the sample adding well; Determine detection information generated by the detection strip according to the detection sample.

20. The method for using the multi-link detection device according to claim 19, wherein: The test sample is a whole blood sample, a plasma sample, or a serum sample.

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