Application of Biological Sample Diluent and Its Detection Method in Immunoassay Kit

Abstract

The present invention belongs to the technical field of biological detection, and specifically relates to the application of a biological sample diluent and its detection method in an immunoassay kit. The biological sample diluent is composed of 6.055 g / L of tris(hydroxymethyl)aminomethane, 35 g / L of sodium chloride, 0.6 mL / L of Triton X-100, 0.6 g / L of surfactant S-9, 1 g / L of bovine serum albumin, and 0.2 mL / L of a liquid biological preservative. The surfactant S-9 and Triton X-100 in the biological sample diluent have a synergistic effect at specific concentrations, effectively improving the foaming phenomenon while also having a high elution and dissolution rate for the target protein, thereby improving the accuracy of immunoassay.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological detection, and particularly relates to the application of a biological sample diluent and its detection method in an immunoassay kit. Background Art

[0002] The IVD product is a combined detection kit for total IgE and eosinophil cationic protein (ECP) (colloidal gold method), which uses the principle of colloidal gold immunochromatographic double antibody sandwich method to qualitatively detect total IgE and eosinophil cationic protein (ECP) in human nasal swabs. The product uses a nasal swab to collect human nasal secretions, elutes and dissolves the collected components carried on the nasal swab in the diluent through the sample diluent, and qualitatively detects the diluent.

[0003] In this product, the main uses of the sample diluent are: (1) eluting the sample collected by the nasal swab to fully separate the sample from the swab and dissolve it in the diluent; (2) providing a stable buffer environment to ensure a stable detection process, uniform chromatography, and ensure the accuracy of detection; (3) protecting the target protein to keep its structure and properties stable during the elution and dissolution process.

[0004] The original diluent of the product consists of 6 components: tris(hydroxymethyl)aminomethane (Tris Base), sodium chloride (NaCl), Triton X-100, surfactant S-9 (Tetronic1307), bovine serum albumin (BSA), and liquid biopreservative (ProClin 300). Among them, Tris Base and NaCl are buffer salt components that provide a pH buffering effect; Triton X-100 and surfactant S-9 are surfactant components that provide the function of promoting elution and dissolution; BSA and ProClin 300 are protein protectants that provide the function of protein stabilization and protection. The above 6 components are dissolved in purified water in a specific ratio, and the content of each component is shown in the formula of the original diluent in Table 1.

[0005] During the use of the original diluent, there is a phenomenon of easy foaming ( Figures 1 - 2 ), and a large amount of foam is generated during use, resulting in bubbles and a long retention time when it is dropped into the sample addition hole of the test card, seriously hindering the sample from entering the conjugate pad and extending upward, causing the risk of inaccurate sample addition amount and abnormal detection results. Such as Figure 3As shown by the foaming situation of the original diluent before and after shaking, 1 mL of the existing diluent was added to the sampling tube, and the intelligent rotary shaking mixer (model MIX-S) was used in vibration mode F8 with a SPEED of 50 for shaking for 30 seconds. After standing for 30 seconds, obvious foam appeared in the tube, and the foam thickness was about 14.5 mm. This foaming situation will have a certain impact on the subsequent sample addition and detection process, making the sample addition volume and detection results inaccurate and causing result deviation.

[0006] In view of the above problems, it is urgent to provide an optimized formula for the diluent to reduce the foaming phenomenon and reduce the detection risk. Summary of the Invention

[0007] In order to solve the above problems, the present invention provides a biological sample diluent, which has less foaming, can effectively elute and dissolve the target protein, and improve the accuracy and sensitivity of detection.

[0008] On the one hand, the present invention provides a biological sample diluent, comprising the following components: tris(hydroxymethyl)aminomethane 6.055 g / L, sodium chloride 35 g / L, Triton X-100 0.6 mL / L, surfactant S-9 0.6 g / L, bovine serum albumin 1 g / L, and biological preservative 0.2 mL / L.

[0009] Specifically, the biological preservative can be selected from one or more of ProClin 300, benzoic acid, sorbic acid, methyl paraben, ethyl paraben, propyl paraben, butyl paraben, sodium dehydroacetate, sodium diacetate, and nisin.

[0010] More specifically, the biological preservative can be selected from one of ProClin 300, benzoic acid, sorbic acid, methyl paraben, ethyl paraben, propyl paraben, butyl paraben, sodium dehydroacetate, sodium diacetate, and nisin.

[0011] Preferably, the biological preservative can be ProClin 300.

[0012] Specifically, the biological sample includes but is not limited to: nasal swab, oral swab, throat swab, cervical swab, male swab, blood, urine or feces.

[0013] Preferably, the biological sample can be a nasal swab, oral swab, throat swab, cervical swab or male swab.

[0014] More preferably, the biological sample can be a nasal swab, oral swab or throat swab.

[0015] Even more preferably, the biological sample can be a nasal swab.

[0016] On the other hand, the present invention provides the use of the aforementioned biological sample diluent in the preparation of an immunoassay kit.

[0017] Specifically, the immunoassay is performed by a colloidal gold immunochromatographic double antibody sandwich method to detect biological samples.

[0018] More specifically, the detection is to detect immunoglobulin E and / or eosinophil cationic protein in biological samples.

[0019] Preferably, the detection is to detect immunoglobulin E and eosinophil cationic protein in biological samples.

[0020] On the other hand, the present invention provides a kit, which includes the aforementioned biological sample diluent.

[0021] Specifically, the kit further includes, but is not limited to, one or more of an enzyme-linked immunosorbent assay (ELISA) plate, a standard, an enzyme-labeled antibody, a washing solution, or a termination solution.

[0022] On the other hand, the present invention provides a detection method, which includes the aforementioned biological sample diluent or kit.

[0023] Technical effects achieved by the present invention:

[0024] (1) The technical solution provided by the present invention can effectively improve the foaming phenomenon.

[0025] (2) The technical solution provided by the present invention has no influence on the elution and dissolution ability of the target protein.

[0026] (3) The technical solution provided by the present invention can meet the technical requirements of a dual-target joint detection kit.

[0027] (4) The technical solution provided by the present invention greatly improves the accuracy of the joint detection kit in the qualitative detection of allergic rhinitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figures 1 - 2 Shows the detection situation of the original diluent that is prone to foaming during the use of the IgE and ECP joint detection kits respectively.

[0029] Figure 3 Shows the foaming situation of the original diluent before and after shaking.

[0030] Figure 4 Shows the foam thickness of Comparative Example 1.

[0031] Figure 5 Shows the foam thickness of Example 1 and Comparative Example 2.

[0032] Figure 6For the foam thickness of the diluent for different components, where T represents "Triton X-100", S represents "surfactant S-9", and B represents "BSA". Detailed implementation manners

[0033] The following combines specific embodiments to further elaborate on the present invention. The following embodiments are not used to limit the present invention, but only to illustrate the present invention. The experimental methods used in the following embodiments, unless otherwise specified, and the experimental methods without specific conditions noted in the embodiments are usually carried out under conventional conditions. The materials, reagents, etc. used in the following embodiments, unless otherwise specified, can all be obtained from commercial channels.

[0034] Example 1

[0035] This example provides a new diluent formulation. Compared with the original diluent formulation, both Triton X-100 (i.e., Triton X-100) and S-9 (i.e., surfactant S-9) are adjusted to 60% (i.e., Triton X-100 is 0.6 mL / L and S-9 is 0.6 g / L). The formulations of Example 1 and the original diluent are shown in Table 1.

[0036] Table 1

[0037]

[0038] Comparative Example 1 Single-component adjustment

[0039] The difference from the original diluent in Example 1 is that the single components of Triton X-100, surfactant S-9, and BSA are respectively adjusted to 80%, 60%, 40%, 20%, and 0% of the corresponding components of the original diluent. Other components are the same as the original diluent in Example 1.

[0040] Comparative Example 2 Two-component adjustment

[0041] On the basis of the original diluent in Example 1, the two surfactants Triton X-100 and S-9 are adjusted in three ratios of 80%, 60%, and 40% respectively (except in Example 1, that is, the combination of 60% S-9 and 60% Triton X-100). Other components are the same as the original diluent in Example 1.

[0042] Effect Example 1

[0043] Measure 1 mL of each adjusted diluent ratio and place it in the product diluent tube. Use the intelligent rotary vibration mixer (model MIX-S) in F8 vibration mode and SPEED 50 for oscillation for 30 seconds. After standing for 30 seconds, use a vernier caliper to measure the foam thickness. The final result is the average value of 2 repeated experiments.

[0044] (1) Comparative Example 1

[0045] The foam thickness of Comparative Example 1 is shown in Table 2 and Figure 4 .

[0046] Table 2 Foam Thickness of Comparative Example 1

[0047]

[0048] Note: The values in the table are the foam film thickness, in millimeters (mm); compared with the original diluent, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

[0049] As Figure 4 can be seen, after the proportion of each of the three diluent components was adjusted separately, the foaming phenomenon was improved. Among them, when the dosage of surfactant S-9 was reduced, the foaming effect was improved most significantly. Since surfactant S-9 plays a role in promoting protein elution and dissolution in the diluent, the elution and dissolution ability of the adjusted formula for the target protein was tested.

[0050] (2) Example 1 and Comparative Example 2

[0051] The foam thickness of Example 1 and Comparative Example 2 is shown in Table 3:

[0052] Table 3 Foam Thickness of Example 1 and Comparative Example 2

[0053]

[0054] Note: The values in the table are the foam film thickness, in millimeters (mm); compared with the original diluent, * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.

[0055] As Figure 5 can be seen, when both Triton X-100 and surfactant S-9 are at a 60% ratio, the diluent has an obvious improvement effect on the foaming effect, and the foam thickness is the smallest (8.44 mm) among all the test combinations. Subsequently, the elution and dissolution ability of this formula diluent was tested.

[0056] Figure 6 For the integrated graph of the single-component adjustment of Comparative Example 1, the foam thickness of Example 1, and the two-component adjustment of Comparative Example 2, it can be clearly observed from the graphical results that when the proportion of the two components is adjusted to 60% of the original diluent for both Triton X-100 and surfactant S-9, the foam formation of the diluent is significantly improved, and the interference of bubbles on the IVD test results is successfully reduced.

[0057] As can be seen from Table 2, when Triton X-100 is 60%, the thickness of the diluted foam decreases by 1.8 mm compared to the original; when surfactant S-9 is 60%, the thickness of the diluted foam decreases by 2.92 mm compared to the original; while when both Triton X-100 and surfactant S-9 in Table 3 are at 60%, the thickness of the diluted foam decreases by 6.07 mm compared to the original. This shows that when both Triton X-100 and surfactant S-9 are at 60%, they have a synergistic effect in reducing the foam thickness.

[0058] Effect Example 2

[0059] According to the results of Effect Example 1, elution and dissolution ability tests were carried out on 20% S-9 and 0% S-9 with good foaming improvement effect in Comparative Example 1, Example 1 (i.e., 60% S-9 and 60% Triton X-100), and 60% S-9 and 40% Triton X-100, 40% S-9 and 60% Triton X-100, 40% S-9 and 40% Triton X-100 in Comparative Example 2.

[0060] An ECP protein ELISA quantitative detection kit was selected to test the elution and dissolution ability of different formulation diluents. The information of the ECP protein ELISA quantitative detection kit is as follows: Name: Human Eosinophil Cationic Protein (ECP) ELISA Kit, Manufacturer: Wuhan Huamei Biological Engineering Co., Ltd., Detection Range: 1.56 ng / mL - 100 ng / mL, Sensitivity 0.39 ng / mL; Catalog Number: CSB-E11729h, Specification 96T / 48T.

[0061] Standard Solution: Dissolve the ECP protein to a concentration of 1 μg / mL, pipette 20 μL and directly add it to 1 mL of the original diluent as the reference standard solution. The theoretical concentration of the standard solution is 20 ng / mL.

[0062] The operation process for the remaining groups is as follows: Pipette 20 μL of the 1 μg / mL ECP solution and drop it onto the nasal swab head. After the liquid is fully absorbed, immerse the swab head into each group of diluents (1 mL) and mix well.

[0063] The concentration points of the standard curve are: 1.56 ng / mL, 3.12 ng / mL, 6.256 ng / mL, 12.5 ng / mL, 25 ng / mL, 50 ng / mL, 100 ng / mL.

[0064] (1) Comparative Example 1

[0065] The experimental groups of Comparative Example 1 were divided into: the original dilution solution group, the 20% S-9 group, and the 0% S-9 group. Each group was set with 5 replicated samples.

[0066] The curve fitting equation is:

[0067] In the formula: A = 4.40332; B = -1.29288; C = 122.06866; D = 0.08976; R 2 = 0.99961.

[0068] The experimental results are shown in Table 4:

[0069] Table 4

[0070]

[0071] Note: Recovery rate = mean concentration of the experimental group / mean concentration of the standard solution × 100%.

[0072] It can be seen from the results in Table 4 that after reducing the proportion of surfactant S-9, the elution and dissolution efficiency of the target protein decreased to a certain extent. The recovery rate of the original ratio dilution solution was 98.5%, the recovery rate of 20% S-9 was 87.6%, and the recovery rate of 0% S-9 was 84.7%. Reducing the proportion of surfactant S-9 alone can improve the foaming phenomenon of the dilution solution to a certain extent, but its elution ability for the target protein is weakened to a certain extent. Therefore, adjusting the components of a single dilution solution cannot take into account both improving the foaming problem and not affecting the elution ability.

[0073] (2) Example 1 and Comparative Example 2

[0074] The experimental groups of Example 1 and Comparative Example 2 were divided into: Example 1 (i.e., 60% S-9 and 60% Triton X-100), 60% S-9 and 40% Triton X-100, 40% S-9 and 60% Triton X-100, 40% S-9 and 40% Triton X-100. Each group was set with 5 replicated samples.

[0075] The curve fitting equation is:

[0076] In the formula: A = 4.22206; B = -1.25372; C = 125.03421; D = 0.07735; R 2 = 0.99947.

[0077] The experimental results are shown in Table 5:

[0078] Table 5

[0079]

[0080] Note: Recovery rate = Mean concentration of experimental group / Mean concentration of standard solution × 100%.

[0081] From the experimental results, it can be seen that the formulated diluent of Example 1 (both Triton X-100 and S-9 are 60%) has no effect on the elution and dissolution ability of the target protein, and the recovery rate is 98.2%, which shows no difference from the recovery rate of 98.5% of the original diluent.

[0082] Effect Example 3

[0083] Select an IgE protein ELISA quantitative detection kit to test the elution and dissolution ability of the selected formulated diluent (both Triton X-100 and S-9 are 60%). Among them, the human immunoglobulin E (IgE) enzyme-linked immunosorbent assay kit is from Beijing Boao Sen Biotechnology Co., Ltd., with a detection range of 468.75 - 30000 pg / mL, product number: BSK1158, and specification: 96T / box.

[0084] Standard solution: Dissolve the IgE protein to a concentration of 500 ng / mL, pipette 20 μL, and directly add it to 1 mL of the original diluent as the reference standard solution. The theoretical concentration of the standard solution is 10 ng / mL.

[0085] The operation process for the remaining groups is as follows: Pipette 20 μL of the 500 ng / mL IgE solution and drop it onto the nasal swab head. After the liquid is fully absorbed, immerse the swab head into each group of diluents (1 mL) and mix well.

[0086] The concentration points of the standard curve are: 0.02 ng / mL, 0.05 ng / mL, 0.10 ng / mL, 0.39 ng / mL, 1.56 ngU / mL, 6.25 ng / mL, 12.5 ng / mL.

[0087] The experimental groups are divided into: the original diluent group, and the group where both Triton X-100 and S-9 are 60%. Each group is set with 5 replicated samples.

[0088] The curve fitting equation is as follows:

[0089] In the formula: A = 4.34256; B = -1.31900; C = 5.00045; D = 0.05949; R 2 = 0.99970.

[0090] The experimental results are shown in Table 6:

[0091] Table 6

[0092]

[0093] Note: Recovery rate = mean concentration of the experimental group / mean concentration of the standard solution × 100%.

[0094] From the experimental results, it can be seen that the formula dilution solution (both Triton X-100 and S-9 are 60%) has no significant effect on the elution and dissolution ability of the target protein, and the recovery rate is 98.9%, which has no significant difference from the recovery rate of 99.4% of the original dilution solution.

[0095] In summary, the formula in Example 1 significantly improves the foaming phenomenon of the dilution solution, and this formula has no effect on the elution and dissolution ability of the target proteins ECP and IgE. Therefore, this formula is a dilution solution formula with more excellent performance effects.

[0096] Effect Example 4

[0097] Since the IgE content in nasal secretions is low, small changes in its concentration may significantly affect the measurement results of the detection kit. In order to simultaneously meet the combined detection test of total IgE and ECP for this IVD product, the sensitivity of the colloidal gold test strip, false positive, chromatographic return white time, and gold-labeled antibody release and chromatography were tested. Four formulas were obtained by changing the composition of the dilution solution: Among them, the formulas of Triton X-100 (i.e., Tween 20) and S-9 (i.e., surfactant S-9) are shown in Table 7, and the other components are the same as those of the original dilution solution.

[0098] Table 7

[0099]

[0100] Sensitivity and false positive test method: Use different sample dilution solutions to add negative nasal swab samples to mix into the dilution solution of the reference product, and then use different reference product dilution solutions to dilute the total IgE reference product so that the final concentrations are 1.0 IU / mL, 0.5 IU / mL, 0.25 IU / mL, 0.125 IU / mL, 0.0625 IU / mL, 0 IU / mL (the dilution solution does not add the reference product). Four groups of samples are obtained and tested using a colloidal gold test strip to obtain the sensitivity under four formulas. Observe whether the sample of 0 IU / mL produces a false positive.

[0101] Chromatographic return white time test method: Observe the time required for the chromatographic background to return white while testing the sensitivity and false positive.

[0102] Gold-labeled antibody release and chromatography: During the chromatography process, observe whether the gold-labeled antibody is evenly chromatographed upward after release, and after the chromatographic background returns white, disassemble the test strip detection card to observe whether the gold-labeled antibody on the gold pad is completely released.

[0103] After preparing the four different formulated diluents to be tested as above, the reference product at 800 IU / mL is diluted to 10 IU / mL using Formulation 1, Formulation 2, Formulation 3, and Formulation 4 respectively, and then further diluted from 10 IU / mL to 1 IU / mL, 0.5 IU / mL, 0.25 IU / mL, 0.125 IU / mL, and 0 IU / mL (the diluent without the reference product). The products from Batch 20240501 are used for detection respectively. The sample loading volume for each well is 60 μL. After loading the samples, start timing, and make the following condition records respectively. Each condition is detected three times.

[0104] Observe the chromatographic situation of the test strips at 5, 8, 10, 13, 15, 20, 25, and 30 minutes respectively.

[0105] Record the time required for the background plate to turn white after loading the samples for each formulation.

[0106] Record the sample detection situation for each formulation at 15 - 20 minutes after loading the samples. "+" indicates positive, and "-" indicates negative.

[0107] After 30 minutes of running the plate, remove the upper cover of the reagent card and observe the release situation of the gold conjugate pad.

[0108] The results are shown in Table 8 - Table 12:

[0109] Table 8 Results of Formulation 1

[0110]

[0111] Table 9 Results of Formulation 2

[0112]

[0113] Table 10 Results of Formulation 3

[0114]

[0115] Table 11 Results of Formulation 4

[0116]

[0117] Table 12 Summary of Test Results

[0118]

[0119] The sensitivity and false positive of colloidal gold test strips directly affect the detection and accuracy of colloidal gold reagents. The chromatographic background return time affects the time to observe the results in colloidal gold detection. If the chromatographic return time is too long, the time to observe the results will be postponed accordingly, which is inconsistent with the purpose of rapid detection. Incomplete release of gold-labeled antibodies and uneven chromatography will affect the uneven color of colloidal gold detection strips and poor detection repeatability. According to the above experiments, Formula 1: The dilution of 60% S9 and 60% Triton x100 components has achieved unexpected technical results and achieved the technical requirements of the dual-target combined detection kit.

[0120] Effect Example 5 Accuracy

[0121] The samples were obtained from the clinical trial project of this IVD product approved by the Ethics Committee of Beijing Century Altar Hospital, and the sample collection process complies with the relevant provisions of the "Ethical Review Measures for Biomedical Research of the People's Republic of China". The samples used for testing were taken from 10 patients who were enrolled in the group. The samples were tested immediately after collection. The sampling swabs should be avoided from being contaminated during the sampling process, and the samples should be tested immediately after sampling. If the collected samples are not tested immediately, they can be stored at 2~8℃ for 48 hours or frozen at -20℃ for no more than 72 hours. The frozen samples should be fully thawed and restored to room temperature before testing, and shaken well.

[0122] The 22 specific IgE antibody detection kits involved are from Jiangsu Haooubo Biotechnology Co., Ltd. and Hunan Haooubo Biotechnology Co., Ltd., including: house dust mites, dust mites, artemisia, ragweed, dog epithelium, cat epithelium, Alternaria, eggs, milk, peanuts, soybeans, cod, shrimp, crab, wheat flour, willow, cockroaches, Aspergillus fumigatus (M3), Humulus (W22), cypress (T23), common birch (T3), sycamore (T11), and allergens. Suxie registration number 20202401671, 20202401682, 20202401668, 20202401680, 20202401669, 20202401705, 20202401711, 20202401689, 20202401715, 20202401716, 20202401670, 20202401681, 20202401692, 20202401700, 20202401691, 20202401704, 20202401688, Hunan Medical Equipment Registration No. 20212401291, 20212401294, 20212401290, 20212401258, 20212401293.

[0123] The test method of the IVD product of the present invention is as follows:

[0124] Nasal secretion sampling method:

[0125] 1. Blow your nose clean with a tissue.

[0126] 2. Sampling:

[0127] (1) Carefully open the outer package of the swab, avoiding contact between your hand and the swab tip.

[0128] (2) Tilt your head slightly back, hold the swab by the tail (do not touch the swab tip with your hand), and insert the swab tip into the nasal cavity to a depth of 1 - 1.5 cm.

[0129] (3) Rotate the swab tightly against the nasal cavity for at least 4 turns (stay for no less than 15 seconds).

[0130] (4) Then use the same swab to repeat the above operation on the other nasal cavity.

[0131] (5) Take another swab and repeat the previous sampling operation to finally obtain two nasal swabs.

[0132] Detection steps:

[0133] (a) Unscrew the caps of the total IgE diluent and ECP diluent tubes;

[0134] (b) Immediately place one of the nasal swabs collected for the sample into the total IgE diluent and the other into the ECP diluent. Rotate the swab tip in the liquid for at least 30 seconds to mix well;

[0135] (c) Squeeze the swab tip forcefully through the outer wall of the tube with your hand at least 5 times to ensure that the liquid is fully released and eluted from the swab;

[0136] (d) Break the swab at the break point (in the middle), leaving the swab tip in the diluent;

[0137] (e) Tighten the caps of the total IgE diluent and ECP diluent tubes;

[0138] (f) Take out the test card, open it along the tear line of the aluminum foil bag, and place the test card flat;

[0139] (g) Unscrew the small caps on the caps of the total IgE diluent and ECP diluent tubes, invert them vertically, gently squeeze the tube body, and drop 3 drops of liquid into the corresponding test sample wells of the test card in sequence;

[0140] (h) Let the test card stand for 15 minutes and observe the displayed result.

[0141] Double positive for IgE and ECP can be judged as allergic rhinitis. The results are shown in Table 13 - 1 and Table 13 - 2.

[0142] Table 13 - 1

[0143]

[0144] Table 13-2

[0145]

[0146] According to the results of Effect Example 4 and Effect Example 5, it can be seen that the foaming phenomenon generated by the diluent during the detection process will significantly affect the detection amount, sensitivity, return to white time and chromatography uniformity of the IVD product sample added dropwise, thus having an adverse impact on the accuracy of the final detection result. During the dropping process of the diluent containing 60% S9 and 60% Triton X-100 described in Effect Example 5 for the IVD product, almost no bubbles are generated, and its detection result is highly consistent with the 22 allergen detection data provided by the hospital and the clinical diagnosis results obtained by the doctor through physical examination. The coincidence rate is significantly improved, greatly enhancing the accuracy of the combined detection kit in the qualitative detection of allergic rhinitis.

Claims

1. A method for diluting a biological sample, characterized in that, The dilution method includes dilution using a biological sample diluent; The biological sample diluent consists of the following components: 6.055 g / L of tris(hydroxymethyl)aminomethane, 35 g / L of sodium chloride, 0.6 mL / L of Triton X-100, 0.6 g / L of surfactant S-9, 1 g / L of bovine serum albumin, and 0.2 mL / L of ProClin 300; the biological sample diluent is used for detecting IGE and ECP in biological samples and improving the foaming phenomenon in detection.

2. The dilution method according to claim 1, wherein The biological sample is a nasal swab, oral swab, cervical swab, male swab, blood, urine, or feces.

3. The dilution method according to claim 2, wherein The biological sample is a nasal swab.

4. The dilution method according to claim 1, wherein The detection is performed on the biological sample by the colloidal gold immunochromatographic double antibody sandwich method.

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