HCT detection kit, HCT detection method and application

Abstract

The invention relates to an HCT detection kit, an HCT detection method and application, and belongs to the technical field of biological detection. The kit comprises anti-mouse IgG (immunoglobulin G), bovine serum albumin, polyethylene glycol 8000, ethylenediamine tetraacetic acid disodium salt, lauryl sodium sulfate and polyethylene glycol octyl phenyl ether. The light transmission effect of the to-be-detected sample is remarkably improved through multiple reagent anti-interference measures, and the detection accuracy and stability are improved. In the detection method, 540nm is selected as a detection wavelength, and an optimized light source, a sensor and a dynamic feedback calibration mechanism are combined, so that high precision of a detection result is ensured, a modified layer on the surface of a reaction vessel has a low adsorption effect, sample residues can be reduced, and the repeatability and accuracy of detection are further improved.

Description

Technical Field

[0001] The present invention belongs to the field of biological detection technology, and specifically relates to an HCT detection kit and a method and application for detecting HCT. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance understanding of the overall background of the invention and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] Hematocrit (HCT) refers to the percentage of blood volume occupied by blood cells in whole blood. Anemia patients have lower HCTs than healthy people, which dilutes the concentrations of other markers in the plasma. When measuring serum markers such as C-reactive protein (CRP) in anemic patients, if the HCT is calculated based on the HCT of healthy people, the deviation in plasma volume caused by abnormal HCT can result in falsely low test values. In certain whole blood testing scenarios in the laboratory, the centrifuged plasma and blood cells need to be remixed. This mixing process can also cause changes in HCT, resulting in deviations in test results.

[0004] Common methods for measuring HCT include electrode measurement (conductivity), hemoglobin absorbance (spectrophotometry), and blood cell counting (impedance / light scattering). The electrode method uses the difference in conductivity between red blood cells and plasma to measure whole blood conductivity and estimate HCT. However, this method is affected by plasma electrolyte concentrations, and electrolyte imbalances in patients can lead to biased results, and regular electrode calibration is required. The hemoglobin absorbance method, based on the absorption properties of hemoglobin at specific wavelengths, measures the absorbance of processed hemoglobin and then converts it to HCT based on an empirical relationship between hemoglobin and red blood cell size. However, red blood cell size (MCV) and morphological abnormalities (such as thalassemia) can affect the accuracy of this empirical relationship. Interferences with absorbance, such as hemolysis, lipemia, and hyperbilirubinemia, can also lead to biased results. A more accurate method is blood cell counting, which includes impedance counting (counting red blood cells by measuring changes in electrical resistance as they pass through micropores and calculating HCT based on their volume) and light scattering (using flow cytometry to analyze cell size and complexity and accurately calculate HCT). However, these methods require large equipment, complex sample processing, and long detection times. Summary of the Invention

[0005] In order to improve the deficiencies of the existing technology, the purpose of the present invention is to provide an HCT detection kit and a method and application for detecting HCT, which integrates immunochromatographic technology to dynamically correct HCT and compensate for the impact of HCT differences on marker concentrations.

[0006] In order to achieve the above objectives, the technical solutions of the present invention are as follows.

[0007] First, an HCT detection kit, comprising: anti-mouse IgG, bovine serum albumin (BSA), polyethylene glycol 8000 (PEG 8000), disodium ethylenediaminetetraacetic acid (EDTA-2Na), sodium dodecyl sulfate (SDS) and polyethylene glycol octylphenyl ether (Triton X-100).

[0008] In a second aspect, a method for detecting HCT comprises the steps of: S1. Sodium dodecyl sulfate (SDS), anti-mouse IgG, bovine serum albumin (BSA), polyethylene glycol 8000 (PEG8000), disodium ethylenediaminetetraacetic acid (EDTA-2Na), and polyethylene glycol octylphenyl ether (Triton X-100) are prepared into a reagent solution. A whole blood sample to be tested is added to the reagent solution to obtain a sample to be tested. A detection light source with a wavelength of 520-560 nm and an auxiliary light source with a wavelength of 850-890 nm are used to detect the absorbance A of the sample to be tested. 540 and turbidity I S90 ; S2. Calibrate the lipemia interference coefficient k using multiple lipemia samples, calibrate the absorbance-HCT conversion coefficient a using a standard substance with HCT = 4.0, and obtain the turbidity nonlinear compensation factor b using regression analysis of hyperlipemia samples; S3, substituting the test result obtained in S1 into formula (1), substituting the coefficient obtained in S2 into formula (1), and obtaining the hematocrit of the whole blood sample to be tested; The formula (1) is: , (1).

[0009] In a third aspect, the application of the above-mentioned method for detecting HCT includes: being used to correct the detection results of markers in plasma.

[0010] The beneficial effects of the present invention are: 1. The present invention uses multiple reagent anti-interference measures, such as anti-mouse IgG, BSA, and PEG 8000, to effectively reduce nonspecific antibody binding, protein adsorption, lipid interference, and turbidity interference. It also utilizes the synergistic effect of SDS and Triton X-100 to accelerate red blood cell membrane lysis and clean the reaction dish, significantly improving the light transmittance of the sample to be tested, and enhancing the accuracy and stability of the test.

[0011] 2. The detection method of this invention uses the hemoglobin absorption peak at 540nm as the detection wavelength. This, combined with an optimized light source, sensor, and dynamic feedback calibration mechanism, ensures high-precision detection results. A low-adsorption treatment on the reaction dish surface reduces sample carryover, further improving detection repeatability and accuracy.

[0012] 3. The detection method of this invention uses the 540nm hemoglobin absorption peak as the detection wavelength. Combined with an optimized light source, sensor, and dynamic feedback calibration mechanism, it ensures high-precision detection results. The use of a low-cost infrared LED instead of a laser diode as an auxiliary light source significantly reduces the manufacturing cost of the device without compromising detection accuracy. The modified layer on the reaction dish surface has a low adsorption effect, reducing sample carryover and further improving detection repeatability and accuracy.

[0013] 4. The HCT test results obtained by this invention can be integrated with immunochromatography technology and used to correct the detection results of markers in plasma. Through the algorithm, the impact of HCT differences on marker concentrations is dynamically compensated. This breaks through the limitations of traditional immunochromatography that relies only on a single optical signal or a fixed calibration curve, and significantly improves the detection reliability of complex samples (such as blood from anemic patients and mixed whole blood). BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The accompanying drawings, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0015] Figure 1 This is a schematic structural diagram of the device in Example 1 of the present invention.

[0016] Among them, 1. LED light source; 2. Filter and collimator; 3. Absorbance detection module; 4. Turbidity detection module; 5. Reaction dish; 6. Outer shell. DETAILED DESCRIPTION

[0017] It should be noted that the following detailed descriptions are exemplary and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0018] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations, devices, components and / or combinations thereof.

[0019] One or more embodiments of the present invention provide an HCT detection kit, comprising: anti-mouse IgG, bovine serum albumin (BSA), polyethylene glycol 8000 (PEG 8000), disodium ethylenediaminetetraacetic acid (EDTA-2Na), sodium dodecyl sulfate (SDS), and polyethylene glycol octylphenyl ether (Triton X-100).

[0020] The test kit is designed based on the SDS-Hb assay method, which allows hemoglobin (Hb) in the blood to react with low-concentration sodium dodecyl sulfate (SDS) to generate an SDS-Hb brown-red compound with an absorption peak at 540nm and an absorption valley at 500nm; The added polyethylene glycol octylphenyl ether (Triton X-100) as a non-ionic surfactant can reduce surface tension and work synergistically with SDS to accelerate the lysis of red blood cell membranes, releasing hemoglobin and forming an SDS-Hb complex in less than 3 seconds. It can also prevent lipid aggregation and stabilize turbidity measurements. Anti-mouse IgG can block the binding sites of non-specific antibodies in the sample, effectively reducing the interference of non-specific antibody binding on the test results; Bovine serum albumin (BSA) can block the adsorption of proteins during the reaction and avoid detection errors caused by protein adsorption.

[0021] Polyethylene glycol 8000 (PEG 8000), as a hydrophilic polymer, can produce a steric hindrance effect, inhibit chylomicron fusion, further reduce sample turbidity interference, and improve the clarity and stability of the detection signal.

[0022] Disodium ethylenediaminetetraacetic acid (EDTA-2Na) is used to chelate metal ions to prevent oxidation and maintain Hb stability.

[0023] Optionally, the anti-mouse IgG is goat anti-mouse IgG.

[0024] One or more embodiments of the present invention provide a method for detecting HCT, comprising the steps of: S1. Prepare a reagent solution containing sodium dodecyl sulfate (SDS) anti-mouse IgG, bovine serum albumin (BSA), polyethylene glycol 8000 (PEG8000), disodium ethylenediaminetetraacetic acid (EDTA-2Na), and polyethylene glycol octylphenyl ether (Triton X-100). Add the whole blood sample to the reagent solution to obtain the sample to be tested. Use a detection light source with a wavelength of 520-560 nm and an auxiliary light source with a wavelength of 850-890 nm to detect the absorbance of the sample to be tested. and turbidity ; S2. Calibrate the lipemia interference coefficient k using multiple lipemia samples, calibrate the absorbance-HCT conversion coefficient a using a standard substance with HCT = 4.0, and obtain the turbidity nonlinear compensation factor b using regression analysis of hyperlipemia samples; S3, substituting the test result obtained in S1 into formula (1), substituting the coefficient obtained in S2 into formula (1), and obtaining the hematocrit of the whole blood sample to be tested; The formula (1) is: , (1).

[0025] In the above process, formula (1) is improved from formula (2) by adding variables k, b and I S90 , significantly improving the accuracy of HCT results. Formula (2) is as follows: , (2).

[0026] Optionally, in S1, the concentrations of the components in the reagent solution are: SDS 30-40 mg / mL, Triton X-100 0.5-0.9 vol.%, PEG 8000 30-70 mg / mL, EDTA-2Na 1.0-2.0 mg / mL, and BSA 6-10 mg / mL.

[0027] Optionally, in S1, the ratio of the reagent solution to the whole blood sample is (18~19): (1~2).

[0028] Optionally, in S1, a detection light with a wavelength of 520-560 nm is emitted by an LED light source, and an auxiliary light with a wavelength of 850-890 nm is emitted by an auxiliary light source. The detection light passes through a filter and a collimator to illuminate a reaction vessel containing a sample to be detected, and an absorbance detection module on the opposite side of the reaction vessel receives the transmitted light through the reaction vessel to obtain the absorbance A. 540 , the turbidity detection module on the opposite side of the reaction vessel receives the scattered light of the reaction vessel contents and obtains the turbidity I S90 .

[0029] Optionally, in S1, the sample to be tested is placed in a reaction vessel for absorbance and turbidity detection, wherein the reaction vessel includes a substrate made of polymethyl methacrylate (PMMA), and a polysulfobetaine (PSBMA) modified layer with a thickness of 150 to 250 nm is provided on the contact surface with the sample to be tested, and a polyethylene glycol (PEG) grafted layer is provided between the modified layer and the substrate.

[0030] Optionally, in S2, the method for calibrating the lipid-blood interference coefficient k for multiple lipid-blood samples includes: first setting the value of the lipid-blood interference coefficient k to 1; taking multiple lipid-blood samples with different lipoprotein contents, and measuring their A 540 and I S90; Then use the hematology analyzer to measure the actual HCT of multiple lipid blood samples, and 540 and I S90 Substitute the data into formula (1) to calculate the HCT value; compare it with the actual HCT, and use mathematical methods to adjust k so that the calculated value of formula (1) is close to the actual value.

[0031] Optionally, in S2, the method for calibrating the absorbance-HCT conversion coefficient a includes: first setting the value of the absorbance-HCT conversion coefficient a to 1; taking multiple samples with different HCT levels, and measuring their A 540 and the actual HCT to calculate the a value for each case; then the corrected a is obtained by averaging or fitting.

[0032] Optionally, in S2, the method for obtaining the turbidity nonlinear compensation factor b by regression analysis includes: first setting the value of the turbidity nonlinear compensation factor b to 0; selecting multiple hyperlipidemia samples, and measuring A 540 , I S90 and the true HCT, substitute the corrected k and a into formula (1) to obtain the calculated HCT, and optimize b through mathematical iteration to reduce the error between the true HCT and the calculated HCT.

[0033] Optionally, in S2, the correction is performed multiple times in the order of k, a, and b until the values ​​of k, a, and b no longer change within the set accuracy, thereby obtaining the final correction result.

[0034] One or more embodiments of the present invention provide applications of the above-mentioned method for detecting HCT, including: correction of detection results of markers in plasma.

[0035] Optionally, the HCT value is substituted into the chromatography reagent card master standard curve for calculation.

[0036] Example 1 A device for detecting HCT, such as Figure 1 As shown, it includes an LED light source 1, a filter and a collimator 2, a reaction vessel 5 and an absorbance detection module 3 arranged in a straight line in a housing 6, and a turbidity detection module 4 and the absorbance detection module 3 are arranged in parallel.

[0037] The LED light source 1 adopts a multi-wavelength light source and selects 540nm as the detection wavelength. This wavelength is the absorption peak of hemoglobin, which can achieve efficient detection of hemoglobin. In this embodiment, the LED light source is a dual-wavelength lamp bead, which can convert between 540 nm wavelength and 850 nm wavelength. The 850nm wavelength is used as an auxiliary light source, and the signal-to-noise ratio and calibration light path are optimized through the filter and collimator 2. Under the premise of ensuring detection accuracy, the equipment cost is greatly reduced.

[0038] The auxiliary light source is used to provide an optical signal complementary to the main detection wavelength. By jointly measuring with the detection light source, it can distinguish background interference (such as lipemia, hemolysis) or optical path fluctuations, thereby improving detection accuracy.

[0039] The absorbance detection module 3 includes a silicon photodiode, which is used to detect the transmitted light through the reaction dish 5 at a receiving angle of 0°, convert its optical signal into an electrical signal, measure the SDS-Hb absorbance, and provide a data basis for subsequent absorbance calculation.

[0040] The turbidity detection module 4 includes an avalanche photodiode, which is used to detect the scattered light of the contents of the reaction dish 5 at a receiving angle of 90°, convert its optical signal into an electrical signal, measure the lipemia turbidity, and provide a data basis for subsequent turbidity calculation.

[0041] The reaction vessel 5 includes a base made of polymethyl methacrylate (PMMA), a polyethylene glycol (PEG) grafted layer is provided on the base, and a polysulfobetaine (PSBMA) modified layer is provided on the grafted layer, serving as a contact surface with the sample to be detected.

[0042] The preparation method of the grafting layer includes: soaking a reaction dish 5 in an ethanol solution of benzoin dimethyl ether (photoinitiator) (the concentration of benzoin dimethyl ether is 1wt%) for 1 hour, blowing it dry, and irradiating it with an ultraviolet lamp for 10 minutes to generate active free radicals on the PMMA surface to obtain an activated PMMA reaction dish; preparing a binary solvent with ethanol and water in a volume ratio of 1:1, adding a sulfobetaine monomer to the binary solvent to a concentration of 5wt%, and adding a PEG monomer to a concentration of 3wt%, to obtain a grafting solution; placing the activated PMMA reaction dish in the grafting solution and irradiating it with an ultraviolet lamp for 2 hours to obtain a grafted layer, and then washing it with deionized water and ethanol multiple times for later use.

[0043] The preparation method of the modified layer includes: spraying 10wt% PSBMA solution on the surface of the grafted layer and then UV curing to make the thickness of the modified layer 200±50nm.

[0044] The purpose of modification and grafting is to improve low adsorption and prevent the sample to be tested from adhering to the surface of the reaction dish 5 and affecting the test results. Experimental verification shows that the contact angle of the untreated PMMA reaction dish is 35° and the SDS-Hb residue is 12.5μg / cm 2 After PEG grafting treatment, the contact angle was less than 5° and the residual SDS-Hb content was reduced to 2.1 μg / cm 2 After PSBMA coating treatment, the contact angle was less than 10° and the residual SDS-Hb content was 1.8 μg / cm 2, effectively reducing sample residue on the reaction dish surface, improving test repeatability and accuracy. Furthermore, the PEG coating can be used stably for over 100 cycles at pH 6-8 and temperatures <50°C, reducing operational costs. Specific performance is shown in Table 1.

[0045] Table 1 Performance of reaction vessels

[0046] Example 2 A HCT detection kit includes: anti-mouse IgG, bovine serum albumin (BSA), polyethylene glycol 8000 (PEG8000), disodium ethylenediaminetetraacetic acid (EDTA-2Na), sodium dodecyl sulfate (SDS) and polyethylene glycol octylphenyl ether (Triton X-100); the anti-mouse IgG is selected as goat anti-mouse IgG.

[0047] The method for detecting HCT using the above-mentioned HCT detection kit comprises the steps of: S0, take 320 μL of reagent solution and add it to the reaction dish, perform blank test and obtain background absorbance data; S1. The sampling arm draws 190 μL of reagent solution into the mixing well. 10 μL of the whole blood sample to be tested is then added to the mixing well and mixed to allow full contact between the whole blood and the reagent solution. This initiates the process of red blood cell membrane lysis and SDS-Hb complex formation, thus obtaining the first step reaction liquid. S1.1, take 35 μL of the first step reaction liquid from the mixing well into the reaction dish and perform further reaction. After mixing in the reaction dish, obtain the sample to be tested. Figure 1 The device of Example 1 is obtained by combining the structure of the embodiment 1 for detection. The LED light source 1 is used to emit detection light with a wavelength of 520-560nm, and the auxiliary light source is used to emit auxiliary light with a wavelength of 850-890nm. The detection light passes through the filter and the collimator 2 to illuminate the reaction dish 5 containing the sample to be detected. The absorbance detection module 3 on the opposite side of the reaction dish 5 receives the transmitted light through the reaction dish 5 to obtain the absorbance. , the turbidity detection module 4 on the opposite side of the reaction vessel receives the scattered light of the reaction vessel contents and obtains the turbidity The tip recovers 355 μL of waste liquid (the complex formed after the reaction of the blood sample and the reagent), completing one detection process.

[0048] S2. Calibrate the lipemia interference coefficient k using multiple lipemia samples, calibrate the absorbance-HCT conversion coefficient a using a standard substance with HCT = 4.0, and obtain the turbidity nonlinear compensation factor b using regression analysis of hyperlipemia samples; S3, substituting the test result obtained in S1 into formula (1), substituting the coefficient obtained in S2 into formula (1), and obtaining the hematocrit of the whole blood sample to be tested; , (1).

[0049] The preparation method of the reagent solution in S0 and S1 (taking the preparation of 100 mL of reagent solution as an example) includes: taking 80 mL of deionized water and placing it in a beaker, and performing the following operations in sequence under magnetic stirring: adding 3.8 g of SDS and stirring until completely dissolved; then adding 0.5 mL of Triton X-100 and stirring to mix; then adding 5 g of PEG8000 and stirring until completely dissolved (heating at 37°C to aid dissolution if necessary) and then cooling; then adding 150 mg of EDTA-2Na, adding NaOH solution dropwise to adjust to pH 8.0 to aid dissolution, and stirring to mix; then adding 1 mL of goat anti-mouse IgG stock solution (diluted in the reagent solution at a ratio of 1:100), gently stirring to mix, adding 0.8 g of BSA, and stirring again; then adding deionized water to make the volume 100 mL and filtering through a 0.22 μm filter membrane.

[0050] The coefficient acquisition method in S2 includes a calibration method for the lipemia interference coefficient k, a correction method for the absorbance-HCT conversion coefficient a, and a regression analysis method for the turbidity nonlinear compensation factor b.

[0051] The calibration method of the lipid-blood interference coefficient k includes: first setting the value of the lipid-blood interference coefficient k to 1; taking 200 lipid-blood samples with different lipoprotein contents, and measuring A with the device for detecting HCT in Example 1; 540 and I S90 ; Then use the hematology analyzer to measure the actual HCT of 200 lipid blood samples, and 540 and I S90 Substitute the data into formula (1) and take a=1 and b=0 to calculate the HCT value; compare it with the actual HCT and adjust k using mathematical methods to make the calculated value of formula (1) closer to the actual value; after the correction process mentioned above, the value of k is corrected from 1 to 0.21.

[0052] The calibration method of absorbance-HCT conversion coefficient a includes: first setting the value of absorbance-HCT conversion coefficient a to 1; taking 150 samples with different HCT levels (minimizing lipid-blood interference), measuring A 540 and actual HCT, using A 540 The value of a was divided by the true HCT value to obtain the a value for each case; then the averaging or fitting method was used to obtain the corrected a. After the above correction process, the value of a was corrected from 1 to 2.35.

[0053] The regression analysis method of turbidity nonlinear compensation factor b includes: first setting the value of turbidity nonlinear compensation factor b to 0; selecting 100 hyperlipidemia samples and measuring A 540 , I S90and the true HCT, substitute the corrected k and a into formula (1) to calculate and obtain the calculated HCT, and optimize b through mathematical iteration to reduce the error between the true HCT and the calculated HCT; after the above process, the value of b is corrected from 0 to 0.00015.

[0054] The coefficients obtained are: k=0.21; a=2.35; b=0.00015.

[0055] After obtaining the above coefficients, the test results of the hematology analyzer were analyzed and the statistical analysis data of 200 samples were obtained as shown in Table 2, which shows that formula (1) has high accuracy and can be used for HCT determination.

[0056] Table 2 Statistical analysis data of 200 samples

[0057] The process of obtaining the coefficient is actually to substitute the test result obtained in S1 into formula (1) before obtaining the whole blood sample to be tested, under the premise that a, b and k in formula (1) have been determined, to obtain the HCT result.

[0058] Comparative Example 1 3.8wt% SDS single reagent was used for detection, and the calculation formula was: , obtain HCT data.

[0059] Result Analysis (1) The test results of Example 2 and Comparative Example 1 are shown in Tables 3-1 and 3-2. In Tables 3-1 and 3-2, Sysmex XN-2800 refers to the HCT data obtained using the Sysmex XN-2800 instrument, 3.8% SDS method refers to the HCT data obtained in Comparative Example 1, innovative detection system refers to the HCT data obtained in Example 2, and error refers to the error relative to the HCT data obtained by the Sysmex XN-2800 instrument.

[0060] Table 3-1 Statistical Table 1 of the test results of Example 2 and Comparative Example 1

[0061] Table 3-2 Statistical Table 2 of the test results of Example 2 and Comparative Example 1

[0062] The statistical analysis data of Comparative Example 2 and Example 3 are shown in Table 4.

[0063] Table 4 Statistical analysis data of Table 3

[0064] The correlation between the HCT data obtained by Example 2 (innovative system) and the Sysmex XN-2800 instrument was 0.981 (R 2 =0.962), which is significantly better than the SDS method's 0.847 (R 2 =0.717), the error dropped from an average of 8.32% to 0.20%, a decrease of 97.6%.

[0065] Example 3 Twenty patients were selected and serum and whole blood samples were obtained respectively. The test item was C-reactive protein. Roche cobas c 701 serum was used as the gold standard for the control group. The whole blood samples were measured using immunochromatographic reagents. The results were directly measured without correction. Under normal circumstances, the default HCT value of healthy people is 37%. A regression curve was obtained by tracing the source with the gold standard serum sample. When the test was a whole blood sample, whole blood = the value in the plasma curve / (1-0.37). At the same time, during the test process, the HCT value of each sample will be obtained through the system.

[0066] Corrected whole blood value = value in plasma curve / (1-actual HCT measured value).

[0067] The clinical validation data and analysis of 20 cases using the immunochromatographic platform whole blood detection + HCT correction vs. serum gold standard are shown in Table 5.

[0068] Table 5 Statistics of whole blood + serum sample test results

[0069] The statistical analysis data of Comparative Example 2 and Example 3 are shown in Table 6.

[0070] Table 6 Statistical analysis data of Table 5

[0071] The HCT correction module enables whole blood testing to reach the serum gold standard level, providing laboratory-level accuracy for POCT scenarios.

[0072] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. A HCT detection kit, characterized in that include: Anti-mouse IgG, bovine serum albumin, polyethylene glycol 8000, disodium edetate, sodium lauryl sulfate, and polyethylene glycol octylphenyl ether.

2. The HCT detection kit according to claim 1, wherein The anti-mouse IgG was selected as goat anti-mouse IgG.

3. A method for detecting HCT, comprising the steps of: S1. Sodium dodecyl sulfate (SDS), anti-mouse IgG, bovine serum albumin (BSA), polyethylene glycol 8000 (PEG8000), disodium ethylenediaminetetraacetic acid (EDTA-2Na), and polyethylene glycol octylphenyl ether (Triton X-100) are prepared into a reagent solution. A whole blood sample to be tested is added to the reagent solution to obtain a sample to be tested. A detection light source with a wavelength of 520-560 nm and an auxiliary light source with a wavelength of 850-890 nm are used to detect the absorbance A of the sample to be tested. 540 and turbidity I S90 ; S2. Calibrate the lipemia interference coefficient k using multiple lipemia samples, calibrate the absorbance-HCT conversion coefficient a using a standard substance with HCT = 4.0, and obtain the turbidity nonlinear compensation factor b using regression analysis of hyperlipemia samples; S3, substituting the test result obtained in S1 into formula (1), substituting the coefficient obtained in S2 into formula (1), and obtaining the hematocrit of the whole blood sample to be tested; The formula (1) is: , (1).

4. The method for detecting HCT according to claim 3, wherein: The concentrations of the components in the reagent solution were: SDS 30–40 mg / mL, Triton X–100 0.5–0.9 vol.%, PEG 8000 30–70 mg / mL, EDTA-2Na 1.0–2.0 mg / mL, and BSA 6–10 mg / mL. Alternatively, in S1, the ratio of the reagent solution to the whole blood sample is (18~19):(1~2).

5. The method for detecting HCT according to claim 3, wherein: In S1, an LED light source is used to emit a detection light with a wavelength of 520-560 nm, and an auxiliary light source is used to emit an auxiliary light with a wavelength of 850-890 nm. The detection light passes through a filter and a collimator to illuminate a reaction vessel containing a sample to be detected. The absorbance detection module on the opposite side of the reaction vessel receives the transmitted light through the reaction vessel to obtain the absorbance A. 540 , using the turbidity detection module on the opposite side of the reaction vessel to receive the scattered light of the reaction vessel contents, and obtain the turbidity I S90 .

6. The method for detecting HCT according to claim 3, wherein: In S1, the sample to be tested is placed in a reaction dish for absorbance and turbidity detection. The reaction dish is a base made of polymethyl methacrylate, and a 150-250 nm thick polysulfobetaine modified layer is provided on the contact surface with the sample to be tested, and a polyethylene glycol grafted layer is provided between the modified layer and the base.

7. The method for detecting HCT according to claim 3, The method is characterized in that, in S2, the method for calibrating the lipemia interference coefficient k of multiple lipemia samples includes: First, set the value of the lipidemia interference coefficient k to 1; Take several blood lipid samples with different lipoprotein contents and measure their A 540 and I S90 ; Then use a hematology analyzer to measure the actual HCT of the lipid blood sample, and 540 and I S90 Substitute the data into formula (1) to calculate the HCT value; compare it with the actual HCT, and use mathematical methods to adjust k so that the calculated value of formula (1) is close to the actual value.

8. The method for detecting HCT according to claim 3, The method of correcting the absorbance-HCT conversion coefficient a in S2 includes: first setting the value of the absorbance-HCT conversion coefficient a to 1; taking multiple samples with different HCT levels and measuring their A values ​​respectively; 540 and the actual HCT, and calculate the a value for each case; then use the averaging or fitting method to obtain the corrected a; Alternatively, in S2, the method for obtaining the turbidity nonlinear compensation factor b by regression analysis includes: first setting the value of the turbidity nonlinear compensation factor b to 0; selecting multiple hyperlipidemia samples, and measuring A 540 , I S90 and the true HCT, substitute the corrected k and a into formula (1) to obtain the calculated HCT, and optimize b through mathematical iteration to reduce the error between the true HCT and the calculated HCT.

9. A use of the method for detecting HCT according to any one of claims 3 to 8, characterized in that: Used to correct the test results of markers in plasma.

10. The use according to claim 9, characterized in that: Substitute the HCT value into the main standard curve of the chromatography reagent card for calculation.

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