Multi-item joint detection kit for xerophthalmia

Abstract

The invention relates to the technical field of medicine and biology, and discloses a xerophthalmia multi-item joint detection kit which is characterized in that the kit is composed of a detection plate, sample treatment liquid, disposable tear detection filter paper strips and a drying agent, the detection plate comprises at least one detection reagent strip, and the sample treatment liquid is arranged in the detection reagent strip. The detection reagent strip is composed of a nitrocellulose membrane, a sample pad, a reagent pad, absorbent paper and a PVC (polyvinyl chloride) board, the reagent pad is a marking solution prepared by respectively marking an MMP-9 antibody, an IgE antibody and an LF antibody by a nanocarbon method, the marking solution is spread on glass fibers after being diluted by a diluent, and the glass fibers are dried overnight at 37 DEG C. According to the invention, the problems of instability and low sensitivity of colloidal gold detection products in the current market are solved. The kit is simple to operate, high in specificity during detection, high in sensitivity and visual in result, and is suitable for home self-test.

Description

Technical Field

[0001] The invention relates to the fields of medicine and biotechnology, and in particular to a multiple combined detection kit for dry eye. Background Art

[0002] Dry eye, also known as keratoconjunctivitis sicca, is a multifactorial disease caused by tears and the ocular surface. The disease is considered to be a disorder of the tear gland functional unit. A complete tear gland unit system includes: tear glands, ocular surface (cornea, conjunctiva and meibomian glands), eyelids, and the sensory and motor nerves connecting them. The clinical manifestations of dry eye are dry eyes, followed by foreign body sensation in the eyes, stinging sensation in the eyes, itchy eyes, photophobia, vision fluctuations, blurred vision, etc. Its pathological mechanisms are mainly increased tear osmotic pressure, tear film instability, ocular surface damage, abnormal nerve sensation and ocular surface inflammation.

[0003] Matrix metalloproteinases (MMPs) are proteolytic enzymes produced by stressed epithelial cells on the ocular surface. In particular, MMP-9 is a nonspecific inflammatory marker. The normal level of MMP-9 in human tears is 3-40 ng / mL, which is continuously elevated in the tears of patients with dry eye. Therefore, MMP-9 can be used as an inflammatory marker for the diagnosis of dry eye disease.

[0004] Tear lactoferrin (LF) as a biomarker has long been established as a "medical standard" for evaluating the secretory function of the tear glands. Tear LF is mainly produced by the lacrimal glands, and a small part comes from the accessory lacrimal glands. It is abundant in tears and together with tear lysozyme, tear prealbumin, and a small amount of secretory immunoglobulins and growth factors, it constitutes the main component of protein in tears. Tear deficiency leads to increased tear film osmotic pressure and reduced lactoferrin levels. The normal content of lactoferrin in tears is between 1.14 and 1.78 mg / mL. The more severe the symptoms of dry eye, the lower the LF content in the tears. Therefore, LF can be used as one of the auxiliary examination methods with strong specificity for diagnosing dry eye, and it is also an indicator for judging its severity.

[0005] Allergic conjunctivitis causes redness, itching, and tearing, which are similar to dry eye symptoms, but the treatment is different from dry eye disease. IgE antibodies are a very important class of immunoglobulins in the human body. Under normal circumstances, their content is the lowest of all immunoglobulins. However, in pathological conditions, such as allergic diseases, IgE levels can be significantly increased. The normal level of IgE in tears is 40ng / ml (1IU=2.4ng / ml). Testing the total IgE level in tears can help clinically diagnose the cause of dry eyes and allergic conjunctivitis. IgE assessment of whether an allergic reaction occurs is very important in the initial dry eye evaluation.

[0006] The main clinical examination methods for dry eyes include tear secretion test (Shirmer test), tear breakup time (BUT), tear fern test, corneal epithelial staining, impression cytology, lysozyme content, lactoferrin content, etc. Among them, the first two tests are relatively quick and simple methods. However, these two methods also have their own shortcomings. The Shirmer I test places the test filter paper into the intraocular fornix, which is difficult to avoid stimulation from some external factors, and the tear secretion volume is unstable and has poor repeatability. The BUT test has high sensitivity, but its specificity is low, and there are also problems with standard determination similar to the Shirmer I test. Summary of the invention

[0007] The main purpose of the present invention is to solve the technical problems of unstable detection and low sensitivity in the prior art. A multiple combined detection kit for dry eye syndrome, the kit is composed of a detection plate, a sample processing liquid, a disposable tear detection filter paper strip, and a desiccant, the detection plate includes at least one detection reagent strip, the detection reagent strip is composed of a nitrocellulose membrane, a sample pad, a reagent pad, a water-absorbing paper, and a PVC board, and the reagent pad is prepared by the following preparation method:

[0008] Disperse the nanocarbon with an ultrasonic device, add it into a centrifuge tube, add the prepared EDC and NHS in sequence, and activate at 35-39℃ for 10-60min;

[0009] After activation, add the antibody dissolved in the coupling solution, mix well, and couple at 35-39℃ for 10-60min;

[0010] After coupling, add nanocarbon blocking solution and emulsifier in sequence, mix well, and block at 35-39°C for 10-60 min;

[0011] After blocking, centrifuge, remove the supernatant and add nanocarbon preservation solution for washing;

[0012] Centrifuge again, remove the supernatant and add nanocarbon preservation solution to obtain labeled nanocarbon solution;

[0013] The labeled nanocarbon solution was diluted with 20 times the volume of nanocarbon labeling diluent and then ultrasonically mixed;

[0014] Evenly drop the ultrasonically homogenized liquid onto the glass fiber placed flat on the glass plate to ensure that the glass fiber is evenly and completely soaked. Transfer the laid reagent pad to the grid rack, put it into an oven at 35-39°C to dry overnight, and store it dry to obtain the reagent pad.

[0015] The antibody is an IgE antibody, a MMP-9 antibody, or a LF antibody.

[0016] The nanocarbon sealing solution is 50mM Tris+2% BSA.

[0017] The nanocarbon preservation solution is 50mM Tris+0.5% BSA.

[0018] The nanocarbon labeling diluent is 70mM Tris+1%, BSA+10%, sucrose+5%, trehalose+0.4% TritonX-100, and the pH value is 8.2.

[0019] The sample treatment solution is 10 mM PBS, pH=7.4.

[0020] The disposable tear test filter paper strip is a Schirmer test paper.

[0021] The nitrocellulose membrane is coated with a detection line T line and a quality control line C line parallel to each other, with a spacing of 5 mm, and is dried at 35-39° C. overnight after coating;

[0022] The sample pad treatment solution contains Na2HPO4, NaH2PO4, NaCl, PVP-30, DEAE, Tween-20, Prolin-300, pH=7.4. After the sample pad is treated with the treatment solution, it is dried at 35-39°C overnight.

[0023] The sample pad treatment solution contains Na2HPO4, KH2PO4, sucrose, Tween-20, S9, BSA, DEAE, CaseinNa, pH=7.4. After the sample pad is treated with the treatment solution, it is dried at 35-39°C overnight.

[0024] The present invention has the following beneficial effects:

[0025] 1. This patent adopts a nanocarbon labeling method, which is completely different from the colloidal gold labeling method. Compared with colloidal gold, nanocarbon has obvious advantages. It has high detection sensitivity and good stability. The carbon material is environmentally friendly. The cost of nanocarbon raw materials is lower than that of colloidal gold. The black and white color contrast between nanocarbon and NC film is easier to distinguish, which solves the problem of instability and low sensitivity of colloidal gold detection products on the market.

[0026] 2. The LF test strip in the kit uses the detection principle of immunochromatographic competition. The test result display lines are consistent with those of the MMP-9 test strip and the IgE test strip, which is convenient for customers to directly observe the results. Positive means two gray-black lines, and negative means one quality control line.

[0027] 3. This patented kit is a combination of multiple marker proteins to detect dry eye. The kit contains MMP-9 detection reagent strips, IgE detection reagent strips, and LF detection reagent strips, which can simultaneously detect three pathogenic factors of dry eye and quickly screen the cause of dry eye. The kit is simple to operate, has high specificity and sensitivity during detection, and the results are intuitive, making it suitable for self-testing at home. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the test result of the IgE test strip using the colloidal gold method;

[0029] Figure 2 The test result of IgE test strip;

[0030] Figure 3 This is the test result of MMP-9 test strip;

[0031] Figure 4 This is the test result of LF test strip;

[0032] Figure 5 This is the test result of the real sample. DETAILED DESCRIPTION

[0033] For ease of understanding, the specific process of the embodiment of the present invention is described below:

[0034] The present invention relates to a multiple combined detection kit for dry eye, the kit comprising a detection plate, a sample processing liquid, a disposable tear detection filter paper strip, and a desiccant, the detection plate comprising at least one detection reagent strip, the detection reagent strip comprising a nitrocellulose membrane, a sample pad, a reagent pad, absorbent paper, and a PVC board, and the reagent pad is prepared by the following preparation method:

[0035] Disperse the nanocarbon with an ultrasonic device, add it into a centrifuge tube, add the prepared EDC and NHS in sequence, and activate at 37°C for 10-60 minutes;

[0036] After activation, add the antibody dissolved in the coupling solution, mix well, and couple at 37°C for 10-60 minutes;

[0037] After coupling, add nanocarbon blocking solution and emulsifier in sequence, mix well, and block at 37°C for 10-60 min;

[0038] After blocking, centrifuge, remove the supernatant and add nanocarbon preservation solution for washing;

[0039] Centrifuge again, remove the supernatant and add nanocarbon preservation solution to obtain labeled nanocarbon solution;

[0040] The labeled nanocarbon solution was diluted with 20 times the volume of nanocarbon labeling diluent and then ultrasonically mixed;

[0041] The ultrasonically homogenized liquid was evenly dripped onto the glass fiber placed flat on the glass plate to ensure that the glass fiber was evenly and completely soaked. The laid reagent pad was transferred to the grid rack and placed in an oven to dry overnight at 37°C. The reagent pad was then stored dry to obtain the reagent pad. Specific embodiment one:

[0043] This embodiment is a method for preparing a MMP-9 detection reagent strip in a kit. The operation steps of the detection reagent strip are as follows:

[0044] 1. Preparation of Sample Pad

[0045] (1) Sample pad treatment solution preparation: Na2HPO4 0.58%, KH2PO4 0.052%, sucrose 1%, Tween-200.05%, S9 1.1%, BSA 1%, DEAE 1.5%, CaseinNa 0.2%. Weigh the reagents according to this formula and add purified water to dissolve. Adjust the pH to 7.4±0.1 with 1 mol / L HCl or NaOH;

[0046] (2) Sample pad preparation: Measure 40 mL of sample pad treatment solution and evenly add it onto glass fiber 8964 (254 mm*300 mm) placed flat on a glass plate. Roll it back and forth 4-5 times with a roller to ensure that the glass fiber is evenly and completely soaked. Transfer the laid sample pad to a grid rack and place it in an oven at 37°C to dry overnight. Store it dry.

[0047] 2. Preparation of Reagent Pad

[0048] Use an ultrasonic device to disperse the nanocarbon, and take 300ul of nanocarbon into a 1.5ml centrifuge tube;

[0049] (1) Add 50 μL of freshly prepared 0.8 mg / mL EDC and NHS in turn, and activate at 37°C for 30 min;

[0050] (2) Prepare the antibody working solution by adding 40 μg of MMP-9 antibody to 100 μL of coupling solution (0.1 M MES);

[0051] (3) Add the antibody working solution to the activated nanocarbon solution, mix well, and couple at 37°C for 30 min;

[0052] (4) After coupling, add 300ul of nanocarbon blocking solution (50mM Tris+2% BSA), 10%

[0053] Tween 100ul, mix well, and block at 37℃ for 30min;

[0054] (5) After blocking, centrifuge at 13000 r / min for 15 min, remove the supernatant and add 800 μL of nanocarbon preservation solution (50 mM Tris + 0.5% BSA) for washing;

[0055] (6) Centrifuge again at 13000 r / min for 15 min, remove the supernatant and add 200 μL of nanocarbon preservation solution for storage.

[0056] (7) Add 10 times the volume of nanocarbon labeling diluent (70 mM Tris + 1% BSA + 10% sucrose + 5% trehalose + 0.4% TritonX-100, PH = 8.2) to the nanocarbon solution labeled in the previous step, dilute and mix by ultrasonication.

[0057] (8) Take 2 ml of the above ultrasonically homogenized liquid and drip it evenly onto the glass fiber KB50 (8 mm*10 mm) placed flat on the glass plate. Roll it back and forth 4-5 times with a roller to ensure that the glass fiber is evenly and completely soaked. Transfer the laid reagent pad to the grid and put it in an oven at 37°C to dry overnight and store it dry.

[0058] 3. Preparation of NC membrane

[0059] (1) C-line coating working solution, coating concentration 1.0 mg / mL, dilute goat anti-mouse with membrane diluent to 1.0 mg / mL, mix well and set aside;

[0060] (2) T-line coating working solution, coating concentration 0.3 mg / mL, dilute MMP-9 antibody to 0.3 mg / mL with membrane diluent, mix well and set aside;

[0061] (3) Adhesive film: PVC board 60mm*280mm, tear off the protective paper in the middle, and stick the NC film along the upper edge of the protective paper below;

[0062] (4) Dot film: Use a gold spray film streaking instrument to coat the C-line antibody and T-line antibody on the NC film fixed on the bottom plate, with an interval of 5 mm, streaking liquid volume: 1 μl / cm; streaking speed: 60 mm / s;

[0063] (5) Dry the scored PVC board at 37°C overnight, add desiccant and seal for later use.

[0064] 4. Assembly of large cards: Cut the dried sample pad into 20mm wide pieces and the reagent pad into 5mm wide pieces;

[0065] First, stick the absorbent paper on the coated NC membrane, and then stick the reagent pad MMP-9 (5 mm wide) and sample pad (20 mm wide) in order;

[0066] 5. Cutting the MMP-9 test strips. After forming the large plate, cut it into 3-4 mm wide test strips, which are the MMP-9 test strips. Specific embodiment 2:

[0068] This embodiment is a method for preparing an IgE detection reagent strip in a kit.

[0069] The preparation method of the reagent strip is the same as that of the MMP-9 detection reagent strip, except for the preparation of the sample pad treatment solution, the preparation of the reagent pad and the NC membrane coating concentration.

[0070] 1. IgE test strip sample pad treatment solution

[0071] The sample pad treatment solution is Na2HPO4 0.15%, NaH2PO4 1.45%, NaCl 0.9%, PVP-30 0.5%, DEAE 1%, Tween-20 1%, Prolin-300 0.1%. According to this formula, weigh the reagents and add purified water to dissolve, and adjust the pH to 7.4±0.1 with 1mol / L HCl or NaOH;

[0072] 2. Preparation of IgE test strip reagent pad

[0073] Use an ultrasonic device to disperse the nanocarbon, and take 200ul of nanocarbon (carboxylated multi-walled carbon nanotubes (length) 30-50nm, product number 100280, Nanjing Xianfeng Nanomaterial Technology Co., Ltd.) into a 1.5ml centrifuge tube;

[0074] (1) Add 50 μL of freshly prepared 0.8 mg / mL EDC and NHS in turn, and activate at 37°C for 30 min;

[0075] (2) Prepare antibody working solution by adding 80 μg of IgE antibody to 100 μL of coupling solution (0.1 M MES);

[0076] (3) Add the antibody working solution to the activated nanocarbon solution, mix well, and couple at 37°C for 30 min;

[0077] (4) After coupling, add 300ul of nanocarbon blocking solution (50mM Tris+2% BSA), 10%

[0078] Tween 100ul, mix well, and block at 37℃ for 30min;

[0079] (5) After blocking, centrifuge at 13000 r / min for 15 min, remove the supernatant and add 800 μL of nanocarbon preservation solution (50 mM Tris + 0.5% BSA) for washing;

[0080] (6) Centrifuge again at 13000 r / min for 15 min, remove the supernatant and add 200 μL of nanocarbon preservation solution for storage.

[0081] (7) Add 20 times the volume of nanocarbon labeling diluent (70 mM Tris + 1% BSA + 10% sucrose + 5% trehalose + 0.4% TritonX-100, PH = 8.2) to the nanocarbon solution labeled in the previous step, dilute and mix by ultrasonication.

[0082] (8) Take 4 ml of the above ultrasonically homogenized liquid and drip it evenly onto the glass fiber KB50 (16 mm*20 mm) placed flat on the glass plate. Roll it back and forth 4-5 times with a roller to ensure that the glass fiber is evenly and completely soaked. Transfer the laid reagent pad to the grid and put it in an oven at 37°C to dry overnight and store it dry.

[0083] 3. Preparation of NC membrane

[0084] (1) C-line coating working solution, coating concentration 1.0 mg / mL, dilute the goat anti-mouse membrane diluent to 1.0 mg / mL, mix well and set aside;

[0085] (2) T-line coating working solution, coating concentration 1.0 mg / mL, dilute IgE antibody to 1.0 mg / mL with membrane diluent, mix well and set aside;

[0086] Other operation steps are the same as those of the MMP-9 detection reagent strip. Specific embodiment three:

[0088] This embodiment is a method for preparing a lactoferrin LF detection reagent strip in a kit.

[0089] The preparation method of the reagent strip is the same as that of the IgE test strip. The difference is the preparation of the sample pad treatment solution.

[0090] LF test strip sample pad treatment solution

[0091] The sample pad treatment solution is 0.1M Tris, PVP-30 0.5%, DEAE 1%, Tween-20 1%, Prolin-3000.1%. According to this formula, weigh the reagents and add purified water to dissolve, and adjust the pH to 7.4±0.1 with 1mol / L HCl or NaOH; Specific embodiment four:

[0093] This example is the preparation of an IgE detection reagent strip (colloidal gold method).

[0094] The preparation method of the reagent strip is the same as that of the IgE detection reagent strip in Example 2. The difference is that the preparation method of the reagent pad is the colloidal gold method.

[0095] Preparation of reagent pad for IgE detection reagent strip (colloidal gold method)

[0096] (1) Take 2 ml of 40 nm colloidal gold solution in a 2 ml EP tube, add 24 ul of 0.1 M K2CO3 and mix well;

[0097] (2) Add 20 μg of IgE antibody and mix well;

[0098] (3) Add 40 μl of 10% BSA, mix well, and centrifuge at 13,000 rpm for 15 min twice.

[0099] (4) After centrifugation, discard the supernatant and keep the precipitated colloidal gold labeling solution.

[0100] (5) Take 5ul of the gold-labeled solution from the previous step, dilute it 400 times with pure water, measure the highest absorption peak on a spectrophotometer and record the value. Calculate the OD of the colloidal gold-labeled solution and select OD5 for gold plating.

[0101] (6) Gold plastering.

[0102] Prepare 1 ml of gold-spreading solution (containing 40% gold label diluent, colloidal gold labeling solution and pure water), mix well, and spread evenly on 5cm*8cm KB50 glass fiber to ensure that the glass fiber is evenly and completely soaked. Transfer the laid reagent pad to the grid, mark it, put it in an oven at 37℃ to dry overnight, and store it dry.

[0103] The other operation steps are the same as those for the preparation of the IgE detection reagent strip in Example 2. Specific embodiment five:

[0105] This example is a verification of the quality control products of the detection reagent strips described in Examples 1, 2, 3, and 4.

[0106] See Figure 1-4 , the quality control products of IgE, MMP-9 and LF were graded diluted with sample diluent. The minimum detection concentration of IgE detection reagent strips by colloidal gold method was 10IU, while the minimum detection concentration of IgE detection reagent strips labeled with nanocarbon method was 1IU, which increased the detection sensitivity by 10 times compared with the colloidal gold method;

[0107] The lowest concentration that the MMP-9 test strip can detect is 2ng / mL, and the highest concentration that the LF test strip can detect is 10ng / mL. The test results are as follows:

[0108] Table 1 Sensitivity test results

[0109] Concentration of quality control products 0 5IU / ml 10IU / ml 20IU / ml Colloidal gold IgE G1 G3 G4 G5 Concentration of quality control products 0 0.5IU / ml 1IU / ml 2IU / ml IgE C1 C3 C4 / C4- C5 Concentration of quality control products 0 2ng / mL 5ng / mL 10 ng / mL MMP-9 C1 C3.5 C4- C4+ Concentration of quality control products 0 10 ng / mL 50ng / mL LF C6+ / C7- C3.5+ / C4- C1

[0110] Note: T line intensity ≥ C3.5 / G3.5 indicates a positive test, < C3.5 / G3.5 indicates a negative test Specific embodiment six:

[0112] Real sample testing of the dry eye syndrome multi-combination test kit

[0113] From left to right, the MMP-9 reagent strip, IgE reagent strip, and LF reagent strip are assembled on a three-in-one plastic part, which is a multi-item dry eye combined detection card.

[0114] 1. Sample Collection

[0115] According to the Schirmer Strips tear sample collection method, aspirate the tears to 10 mm, drop the filter paper strips into the sample diluent tube, and mix for 1 minute;

[0116] 2. Detection

[0117] 1) Tear open the aluminum foil bag, take out the test plate from the aluminum foil bag, and place it flat on the table;

[0118] 2) Add 3-4 drops (about 100ul) of the above sample treatment solution to each of the three sample wells of the detection plate;

[0119] 3) Timed observation: The results will be read after 15-20 minutes. Results read after 20 minutes will be invalid.

[0120] 3. Test results

[0121] The experimental samples were self-tested by laboratory staff, 1# was a normal sample; the donors of samples 2# and 3# had dry eyes and ocular oxygen.

[0122] Test results: see Figure 5 The results were consistent with the symptoms provided by the experimental samples, 2# and 3# were allergic conjunctivitis and dry eyes, respectively.

[0123] From the verification of the detection methods of Examples 4 and 5, the multiple combined detection kit for dry eye provided by this patent has simple operation, high sensitivity, and intuitive results. It is a simple, accurate and economical rapid detection method.

[0124] As described above, the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features thereof may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A multiple combined detection kit for dry eye syndrome, characterized in that: The kit is composed of a test plate, a sample processing liquid, a disposable tear test filter paper strip, and a desiccant. The test plate includes at least one test reagent strip, and the test reagent strip is composed of a nitrocellulose membrane, a sample pad, a reagent pad, absorbent paper, and a PVC board. The reagent pad is prepared by the following preparation method: 1 mg of nano-carbon powder was weighed and dissolved and dispersed in 1 ml of 1% carbon nanotube dispersant (purchased from Nanjing Xianfeng Nanotechnology Co., Ltd., item number: 101112). Disperse the nanocarbon with an ultrasonic device, add it into a centrifuge tube, add the prepared EDC and NHS in sequence, and activate at 35-39℃ for 10-60min; After activation, add the antibody dissolved in the coupling solution, mix well, and couple at 35-39℃ for 10-60min; After coupling, add nanocarbon blocking solution and emulsifier in sequence, mix well, and block at 35-39°C for 10-60 min; After blocking, centrifuge, remove the supernatant and add nanocarbon preservation solution for washing; Centrifuge again, remove the supernatant and add nanocarbon preservation solution to obtain labeled nanocarbon solution; The labeled nanocarbon solution was diluted with 20 times the volume of nanocarbon labeling diluent and then ultrasonically mixed; Evenly drop the ultrasonically homogenized liquid onto the glass fiber placed flat on the glass plate to ensure that the glass fiber is evenly and completely soaked. Transfer the laid reagent pad to the grid rack, put it into an oven at 35-39°C to dry overnight, and store it dry to obtain the reagent pad.

2. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The antibody is an IgE antibody, a MMP-9 antibody, or a LF antibody.

3. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The nanocarbon sealing solution is 50mM Tris+2% BSA.

4. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The nanocarbon preservation solution is 50mM Tris+0.5% BSA.

5. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The nanocarbon labeling diluent is 70mM Tris+1%, BSA+10%, sucrose+5%, trehalose+0.4% TritonX-100, and the pH value is 8.

2.

6. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The sample treatment solution is 10 mM PBS, pH=7.

4.

7. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The disposable tear test filter paper strip is a Schirmer test paper.

8. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The nitrocellulose membrane is coated with a detection line T line and a quality control line C line which are parallel to each other and are spaced apart by 5 mm. After coating, the membrane is dried at 35-39° C. overnight.

9. A multiple combined detection kit for dry eye according to claim 1, characterized in that: The sample pad treatment solution contains Na2HPO4, NaH2PO4, NaCl, PVP-30, DEAE, Tween-20, Prolin-300, pH=7.

4. After the sample pad is treated with the treatment solution, it is dried at 35-39°C overnight.

10. The multiple combined detection kit for dry eye according to claim 1, characterized in that: The sample pad treatment solution contains Na2HPO4, KH2PO4, sucrose, Tween-20, S9, BSA, DEAE, CaseinNa, pH=7.

4. After the sample pad is treated with the treatment solution, it is dried at 35-39°C overnight.

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