High-density lipoprotein cholesterol determination kit and detection method thereof

Abstract

The invention discloses a high-density lipoprotein cholesterol determination kit and a detection method thereof. The kit comprises a reagent 1 and a reagent 2, the reagent 1 is prepared from a buffer solution, inorganic salt, dextran sulfate, a macromolecular cationic polymer, TOOS, peroxidase, ascorbic acid oxidase and a preservative; the reagent 2 is prepared from a buffer solution, a surfactant 1, 4-aminoantipyrine, cholesterol oxidase, cholesterol esterase and a preservative. The kit has the advantages of being high in accuracy, good in anti-interference performance and the like, automatic detection can be achieved on a full-automatic biochemical analyzer, and compared with a traditional dextran sulfate precipitation method, the content of high-density lipoprotein cholesterol (HDL-C) in a lipid turbid sample can be more accurately detected.

Description

Technical Field

[0001] The present invention relates to the field of biochemical analysis, and in particular to a high-density lipoprotein cholesterol (HDL-C) determination kit and a detection method thereof. Background Art

[0002] Cholesterol is an important indicator of lipid metabolism in the human body. Only 10% of the cholesterol in the human body comes directly from animal food, and most of the cholesterol is produced by the liver. Cholesterol exists in the human body in two forms: free cholesterol, which accounts for 30% of the total cholesterol; and cholesterol ester, which exists in lipoproteins. Human lipoproteins can be roughly divided into the following four categories: 1. Chylomicrons CM, 2. Very low density lipoprotein VLDL, 3. Low density lipoprotein LDL, 4. High density lipoprotein HDL.

[0003] HDL is the lipoprotein with the highest density among lipoproteins, and is composed of proteins, phospholipids, cholesterol and its esters, and triglycerides. The proteins in HDL are mainly apolipoprotein (APo) AI and AⅡ, and the main lipids are phospholipids, with lecithin being the largest, followed by sphingomyelin and lysophospholipids. Cholesterol is the second largest lipid in HDL, and the ratio of esterified and free cholesterol is about 3:1. HDL has many physiological effects. First, HDL can transfer cholesterol from surrounding tissues to the liver for degradation through the action of enzymes and receptors, while inhibiting cells from binding to and taking up low-density lipoproteins, and preventing cholesterol from depositing in the arterial wall. Therefore, HDL is considered to be a preventive factor for atherosclerosis (AS). Secondly, it can promote endothelial progenitor cell-mediated endothelial repair, that is, it can help repair the endothelial tissue of blood vessels. Thirdly, it can stimulate blood endothelial cells to produce nitric oxide to protect the vascular endothelium. Finally, it can enter the endothelium, regulate the contraction and relaxation of blood vessels, and also have an antioxidant effect. Therefore, the detection of high-density lipoprotein cholesterol (HDL-C) has important clinical significance.

[0004] Traditional methods for testing HDL-C include centrifugation, electrophoresis, and precipitation. The internationally recognized reference method for HDL-C and LDL-C determination is the ultracentrifugation-heparin manganese precipitation-AK cholesterol determination method of the Centers for Disease Control and Prevention (CDC), also known as the β-quantification method. This method is also the only HDL-C and LDL-C reference method listed in the reference method list of the Joint Traceability Committee for Clinical Chemistry (JCTLM). However, this method is cumbersome to operate and has many influencing factors, making it difficult to be widely used. Later, for the determination of HDL-C, the CDC established a modified dextran sulfate precipitation method (DCM), which is widely used in the CDC blood lipid standardization program because of its simplicity and rapidity.

[0005] With the development of technology, HDL-C can be fully automatically detected. According to the reaction principle, the methods currently available for analyzing HDL-C with fully automatic biochemical analyzers include enzyme modification method, immunoseparation method, selective inhibition method, clearance method, etc. However, these methods all have different defects, such as poor specificity and poor anti-lipid turbidity ability.

[0006] The dextran sulfate precipitation method is a reference method for CDC to determine HDL-C. Its principle is to first use dextran sulfate combined with magnesium ions to precipitate non-HDL lipoproteins, centrifuge to remove the precipitate, leaving only HDL-C in the supernatant, HDL-C generates cholesterol under the action of cholesterol esterase, cholesterol generates hydrogen peroxide under the action of cholesterol oxidase, hydrogen peroxide causes the chromogen to develop color under the action of peroxidase, resulting in an increase in absorbance, and the HDL-C content is calculated based on the change in absorbance. This method has a high accuracy. However, when this principle is applied to a fully automatic biochemical analyzer, due to the lack of a centrifugation step, when measuring lipid turbidity samples (chylomicron or high triglyceride TG>1.8mmol / L), reagent 1 reacts with the sample to generate more precipitates, causing a significant increase in absorbance, resulting in a deviation in the absorbance change after the cholesterol reaction, and the result is low (the proportion of the precipitate generated by non-HDL lipoproteins and reagents in the entire reaction volume of healthy human serum is too low, and there will be no significant increase in absorbance). Summary of the invention

[0007] In view of the shortcomings of the prior art, the technical problem to be solved by the present invention is to provide a high-density lipoprotein cholesterol determination kit and a detection method thereof, which can solve the problem of fat turbidity interference in the fully automatic detection of the dextran sulfate precipitation method. The present invention uses a high molecular cationic polymer to quickly aggregate and precipitate the precipitate formed by the fat turbidity sample, so that the reaction system is quickly clarified and the absorbance quickly returns to normal, avoiding interference with subsequent reactions, thereby ensuring the accuracy of the results.

[0008] The technical solution adopted by the present invention to solve the technical problem is as follows:

[0009] A high-density lipoprotein cholesterol (HDL-C) determination kit comprises reagent 1 and reagent 2, wherein the components and concentrations of reagent 1 are:

[0010]

[0011] The components and concentrations of the reagent 2 are:

[0012]

[0013] Furthermore, the high molecular cationic polymer is a polymer that can rapidly aggregate and precipitate the precipitate produced by dextran sulfate and non-HDL lipoprotein from a dispersed state within 3 minutes.

[0014] Furthermore, the buffers in the reagents 1 and 2 are independently selected from: phosphate buffer, 3-[NN-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulfonic acid buffer, glycine buffer, 4-hydroxyethylpiperazineethanesulfonic acid buffer, 3-morpholinepropanesulfonic acid buffer, etc. The pH value of the buffer is 7.0-8.0.

[0015] The inorganic salt in the reagent 1 is selected from one or more of sodium chloride, potassium chloride, magnesium chloride, calcium chloride and the like.

[0016] The dextran sulfate in the reagent 1 is selected from at least one of: dextran sulfate-molecular weight 3000, dextran sulfate-molecular weight 20000, dextran sulfate-molecular weight 40000, dextran sulfate-molecular weight 50000, etc.

[0017] The high molecular cationic polymer in the reagent 1 is selected from at least one of: amine polymers, imine polymers, polyquaternary ammonium salts, etc.;

[0018] The amine polymer is at least one of polyamine and polyethyleneamine; the imine polymer is at least one of polyethyleneimine and polypropyleneimine; the polyquaternary ammonium salt is at least one of polyquaternary ammonium salt-5, polyquaternary ammonium salt-6, polyquaternary ammonium salt-7, and polyquaternary ammonium salt 39.

[0019] Furthermore, the preservatives in the reagent 1 and the reagent 2 are selected from one or more of sodium azide, PC300, gentamicin, sodium benzoate and the like.

[0020] The surfactant 1 in the reagent 2 is selected from one or more of the emulsifiers AEO-3, AEO-7, AEO-9, etc.

[0021] Preferably, the concentration of each component in the kit is:

[0022] Reagent 1 is:

[0023]

[0024] Reagent 2 is:

[0025]

[0026] The pH value of the above buffer is 7.0-8.0.

[0027] In a second aspect, the present invention provides a detection method for a high-density lipoprotein cholesterol determination kit, the process of the method is: after adding reagent 1 to the sample on the biochemical analyzer, incubate for 3 minutes, the high molecular cationic polymer adsorbs dextran sulfate DS and quickly aggregates and precipitates the dextran sulfate and non-HDL lipoprotein from a dispersed state within 3 minutes, so that the absorbance is restored to the initial level, that is, the absorbance level when no precipitation is produced, and then reagent 2 is added.

[0028] Furthermore, the reagent 1 is a reagent containing dextran sulfate and a high molecular cationic polymer, and the interference of the fat turbidity sample is eliminated by precipitation method;

[0029] The reagent 2 is a reagent containing cholesterol oxidase, cholesterol esterase and 4-aminoantipyrine.

[0030] Furthermore, the upper limit of the anti-interference of the kit for chyle samples is increased to more than 2%; for lipid turbidity samples, the deviation of the multiple dilution back calculation of the kit is controlled within 10%.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] The invention adopts a high molecular cationic polymer to quickly aggregate the precipitate produced by dextran sulfate and fat turbidity samples, eliminates the interference caused by the increase of absorbance, solves the problem of low measured value of fat turbidity samples, and overcomes the problem of poor anti-fat turbidity ability when the existing dextran sulfate precipitation method is applied to the full-automatic biochemical analyzer. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The absorbance change curve of the reaction process of measuring HDL-C using the dextran sulfate precipitation method for healthy human samples (DS or DS+PEI);

[0034] Figure 2 The absorbance change curve of the reaction process of measuring HDL-C using the traditional dextran sulfate precipitation method for lipid turbidity samples (only dextran sulfate DS was added);

[0035] Figure 3 The absorbance change curve of the reaction process of measuring HDL-C using the improved dextran sulfate precipitation method of the present application for lipid turbidity samples (adding dextran sulfate DS and polyethyleneimine PEI);

[0036] Figure 4 This is a diagram showing the effect of different molecular weight PEI combined with DS on the detection of lipid turbidity samples.

[0037] Figure 5 It is a comparison chart of the linear test results of Comparative Examples 1 and 2 and Examples 1 and 2.

[0038] Figure 6It is a comparison diagram of the reaction curves of the turbid sample 2 (normal turbidity TG=6.5g / L) and the turbid sample 4 (severe turbidity TG=15.4g / L) in different control examples and embodiments. DETAILED DESCRIPTION

[0039] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the technical solutions of the present invention are described in detail below in conjunction with embodiments.

[0040] The kit of the present invention comprises reagent 1 and reagent 2; reagent 1 comprises buffer, inorganic salt, dextran sulfate, high molecular cationic polymer, TOOS, peroxidase, ascorbic acid oxidase and preservative, but does not contain surfactant; reagent 2 comprises buffer, surfactant 1, 4-aminoantipyrine, cholesterol oxidase, cholesterol esterase and preservative.

[0041] Figure 1-Figure 3 This is a comparison chart of the absorbance change curves of the reaction process of measuring HDL-C using healthy human samples and lipid turbidity samples using the traditional dextran sulfate precipitation method and the dextran sulfate precipitation method improved by the present application. Figure 1-Figure 3 The blue color is the main wavelength, the green color is the secondary wavelength, and the red color is the main wavelength-secondary wavelength. The high molecular cationic polymer used in the improved dextran sulfate precipitation method test process of this application is polyethyleneimine PEI; the test process is to first add reagent 1 and sample, incubate for 3 minutes, read the absorbance A1, then add reagent 2, incubate for 5 minutes, and read the absorbance A2.

[0042] Compared with the reaction curve of healthy human samples, when testing lipid turbidity samples, only DS (dextran sulfate), reagent 1 and sample incubation process were added, and the absorbance increased significantly, which had a great interference with the subsequent absorbance measurement. When DS and PEI were added at the same time, the precipitate formed by reagent 1 and the sample quickly aggregated, and the turbidity turned into clarity, and the absorbance dropped rapidly to the initial level, thus eliminating the interference of lipid turbidity on the final measurement value.

[0043] The basic principle of the present invention is as follows: DS (dextran sulfate) is a negatively charged polysaccharide, which can react with positively charged lipoprotein (non-HDL lipoprotein) in solution to form a precipitate; a high molecular cationic polymer can combine with negatively charged biological molecules to make them quickly aggregate and precipitate. In the present invention, DS is used to capture non-HDL lipoprotein, and then DS is adsorbed by a high molecular cationic polymer, so that the precipitate generated by DS and lipoprotein quickly aggregates and precipitates from a dispersed state (the sample amount used for biochemical instrument detection is very small, and the precipitate after aggregation and sedimentation will not affect the subsequent reaction cup cleaning), which has an effect similar to centrifugation, restores the absorbance to normal, eliminates the interference of turbid samples, and improves the accuracy and anti-turbidity ability of the reagent.

[0044] The high molecular cationic polymer used in the present invention is at least one of amine polymers, imine polymers, polyquaternary ammonium salts, etc.;

[0045] The amine polymer is at least one of polyamine, polyethyleneamine, etc.; the imine polymer is at least one of polyethyleneimine, polypropyleneimine, etc.; the polyquaternary ammonium salt is at least one of polyquaternary ammonium salt-5, polyquaternary ammonium salt-6, polyquaternary ammonium salt-7, polyquaternary ammonium salt 39, etc.

[0046] Since the high molecular cationic polymer has different molecular weights according to the degree of polymerization, the present invention selected different PEI with molecular weights of 600-250000 on the market to cooperate with DS to test the lipid turbidity sample to verify its aggregation precipitation effect. The results are as follows: Figure 4 As shown in the figure above, it can be seen that the larger the PEI molecular weight, the faster the absorbance decreases after the precipitation, indicating that the rate at which PEI causes the precipitation to aggregate and settle is proportional to its molecular weight; on the biochemical analyzer, after adding reagent 1 to the sample and incubating for about 3 minutes, reagent 2 is added. To ensure that the absorbance returns to normal within 3 minutes, PEI with a molecular weight of 3000 or above should be selected (the absorbance of PEI3000 in the figure drops to the bottom at 170s). The preferred PEI molecular weight is 3000-300000. The sedimentation effect of DS and PEI of different molecular weights is not much different and can be ignored.

[0047] For the molecular weight selection of other high molecular cationic polymers, the minimum molecular weight limit may be different according to their different structures, and the minimum molecular weight limit may be based on the ability to quickly aggregate and settle the precipitate produced by dextran sulfate and non-HDL lipoprotein from a dispersed state within 3 minutes.

[0048] Control Example: (Traditional Dextran Sulfate Precipitation Method)

[0049] Control Example 1: Preparation of a high-density lipoprotein cholesterol (HDL-C) assay kit

[0050] Reagent 1 (R1) is:

[0051]

[0052] Reagent 2 (R2) is:

[0053]

[0054] Control Example 2: Preparation of a high-density lipoprotein cholesterol (HDL-C) assay kit

[0055] Reagent 1 (R1) is:

[0056]

[0057] Reagent 2 (R2) is:

[0058]

[0059]

[0060] The main difference between Control Example 2 and Control Example 1 is that Control Example 1 adopts the traditional dextran sulfate precipitation method, while Control Example 2 adds amide cationic polymers on the basis of Control Example 1.

[0061] Example 1: Preparation of a high-density lipoprotein cholesterol (HDL-C) assay kit

[0062] Reagent 1 (R1) is:

[0063]

[0064] Reagent 2 (R2) is:

[0065]

[0066] The main difference between Example 1 and Comparative Example 2 is that the cationic polymer used in Reagent 1 is different.

[0067] Example 2: Preparation of a high-density lipoprotein cholesterol (HDL-C) assay kit

[0068] Reagent 1 (R1) is:

[0069]

[0070] Reagent 2 (R2) is:

[0071]

[0072]

[0073] The main difference between Example 2 and Example 1 is that the molecular weight and amount of dextran sulfate and polyethyleneimine are different.

[0074] 1. The performance of the control example 1 / 2 and the embodiment 1 / 2 was evaluated, and the evaluation index and method were as follows:

[0075] 1.1 Reagent blank absorbance: At 37°C and 600nm wavelength, the reagent blank absorbance should be <0.05.

[0076] 1.2 Analytical sensitivity: When the sample concentration is 1mmol / L, the absorbance difference should be greater than 0.04.

[0077] 1.3 Repeatability: Two serum concentrations, high and low, were tested. Each sample was tested 10 times, and the coefficient of variation (CV) was <4%.

[0078] 1.4 Accuracy: The relative deviation of third-party quality control products shall not exceed ±10%.

[0079] 1.5 Linearity: In the range of [0.08, 3.90] mmol / L, the linear correlation coefficient 2 is ≥ 0.995. In the range of (1.0, 3.90] mmol / L, the relative deviation does not exceed ± 10%, and in the range of [0.08, 1.0] mmol / L, the absolute deviation should not exceed ± 0.1 mmol / L.

[0080] 1.6 Clinical comparison: Test one group of random serum and one group of high triglyceride serum to compare the correlation and deviation between the control sample and the reagents of the embodiment.

[0081] 1.7 Anti-interference performance: Interference sample preparation: Preparation is carried out by adding interfering substances to the serum; the influence of a certain concentration of interfering substances on the test results is considered to have no significant effect within ±10%.

[0082] Types and standards of interference substances: The upper limit of anti-hemoglobin interference is 5g / L; the upper limit of anti-ascorbic acid interference is 0.5g / L; the upper limit of anti-bilirubin interference is 100umol / L; the upper limit of anti-chylomicron interference is 1%; the upper limit of anti-heparin sodium interference is 100IU / L.

[0083] 1.8 Lipid turbidity sample detection: Divide the lipid turbidity sample into two parts, one for control and the other for pretreatment: manually add dextran sulfate and magnesium ions to remove non-HDL lipoproteins in the sample, and then perform fully automatic detection to compare the difference in HDL-C values ​​before and after treatment.

[0084] Lipid sample pretreatment process:

[0085] Experimental materials: 4 cases of lipid turbidity serum (TG>2g / L, TG reference range: 0.4-1.6g / L for males and 0.2-1.08g / L for females), numbered 1-4.

[0086] DS (Dextran Sulfate DS50) precipitation reagent: DS50: 10 g / L, magnesium chloride: 20 g / L.

[0087] Experimental method: 4 samples of turbid sera were treated with 1 ml serum + 1 ml DS precipitation reagent, mixed with DS reagent and allowed to stand for 10 minutes, then centrifuged to obtain the supernatant.

[0088] After pretreatment, lipid-turbid serum before and after treatment was tested simultaneously to compare the differences in HDL-C values.

[0089] 2. Evaluation results:

[0090] 2.1 Blank absorbance and analytical sensitivity

[0091] 2.2 Repeatability

[0092]

[0093] Conclusion: The repeatability of the control example and the embodiment is qualified 2.3 Accuracy

[0094]

[0095] Conclusion: The accuracy of the control example and the embodiment is qualified 2.4 Linearity (results are shown in the table below and Figure 5 )

[0096]

[0097] Conclusion: The linearity of the control example and the embodiment is qualified

[0098] 2.5 Clinical comparison

[0099]

[0100] Conclusion: When testing random serum, the overall measured value of control example 1 / 2 is slightly lower than that of example 1 / 2, but the deviation is small, and the overall deviation of multiple dilution is good; when testing lipid turbid serum (TG high value sample), the measured value of control example 1 / 2 is about 20% lower than that of example 1 / 2, and after multiple dilution, the reverse calculation deviation of control example 1 / 2 is 19.7% and 19.0%, respectively, and the reverse calculation deviation is <10% 2.6 Anti-interference performance

[0101]

[0102] Conclusion: There is no significant difference in the anti-interference ability to hemoglobin, VC, bilirubin and heparin sodium. The anti-interference ability of Example 1 / 2 to chyle samples is significantly improved compared with that of Control Example 1 / 2, and the upper limit of anti-interference is increased from 0.5%-1% to about 2%.

[0103] 2.7 Lipid turbidity sample test (results are shown in the table below and Figure 6 )

[0104]

[0105] Conclusion: Before treatment: the lipid turbidity sample was directly tested, and the result of the control example was significantly lower than that of the embodiment, and from the reaction curve No. 2 / 4, the absorbance of the control example R1 increased significantly after mixing with the sample; while the absorbance of the embodiment increased and then dropped rapidly to return to normal; After treatment: the sample was first treated with dextran sulfate and then tested. At this time, the measured results of the control example and the embodiment were similar, and the first half of the reaction curve of the control example of sample No. 2 / 4 no longer increased, indicating that the difference in the measured values ​​between the control example and the embodiment before sample treatment was caused by the lipid turbidity problem of the sample; The results show that: the embodiment can effectively eliminate the interference of lipid turbidity.

[0106] 3. Conclusion summary:

[0107] 3.1 Basic performance of reagents: blank absorbance, analytical sensitivity, repeatability, linearity, accuracy. There was no significant difference between the control example and the embodiment, and all were qualified.

[0108] 3.2 Clinical comparison: Random human serum was tested. The overall deviation between the control example and the embodiment was small, and the correlation was good. The reverse calculation deviation of individual samples of the control example after multiple dilution was large, while the reverse calculation deviation of the embodiment was small. Turbid serum was tested: the measured values ​​of the control example were lower than those of the embodiment as a whole, and the reverse calculation deviation of the measured values ​​of the control example after multiple dilution was large (>19%), while the reverse calculation deviation of the embodiment was small (within ±5%).

[0109] 3.3 Anti-interference performance: There is no significant difference in the anti-interference ability to hemoglobin, VC, bilirubin, and heparin sodium. The anti-interference ability of the embodiment to chyle samples is significantly improved compared with the control example, and the upper limit of anti-interference is increased from 0.5%-1% to about 2%.

[0110] 3.4 Detection of turbid samples: Before treatment: The turbid samples were directly tested, and the results of the control example were significantly lower than those of the example. From the reaction curve of sample No. 2 / 4, the absorbance of the control example R1 increased significantly after mixing with the sample; while the absorbance of the example decreased rapidly after the increase and returned to normal; After treatment: The sample was first treated with dextran sulfate and then tested. At this time, the measured results of the control example and the example were similar, and the first half of the reaction curve of the control example of sample No. 2 / 4 no longer increased; the measured values ​​of the control example before and after treatment were greatly different, while the measured values ​​of the example changed less; This shows that the difference in the measured values ​​between the control example and the example before sample treatment was caused by the turbidity of the sample.

[0111] Based on the test results of lipid turbidity sample dilution ratio, anti-chylolytic ability and lipid turbidity sample in clinical comparison, it can be determined that compared with the traditional dextran sulfate method or dextran sulfate combined with polyacrylamide method, the polyethyleneimine combined with dextran sulfate method adopted in the present invention can effectively eliminate the interference problem of lipid turbidity samples.

[0112] Example 3

[0113] The components and detection methods of this embodiment are the same as those of Example 1, except that the high molecular cationic polymer added to reagent 1 in this embodiment is one or more of polyamine, polyethyleneamine, polypropyleneimine, polyquaternium-5, polyquaternium-6, polyquaternium-7 or polyquaternium 39, to replace the polyethyleneimine-molecular weight 10000 in Example 1. The molecular weights are all above 8000.

[0114] Example 4

[0115] The components and detection methods of this embodiment are the same as those of Example 1, except that in this embodiment, the polyethyleneimine is selected as a mixture of molecular weights of 3,000 and 170,000 or 250,000, or a mixture of molecular weights of 600 and 100,000 or 250,000, so that it can quickly aggregate and precipitate the precipitate produced by dextran sulfate and non-HDL lipoprotein from a dispersed state within 3 minutes, so that the absorbance returns to a normal level.

[0116] The embodiments of the present application can achieve the purpose of eliminating the interference of turbid samples by changing different types of high molecular cationic polymers, which is an attempt at a new method and has significant progress.

[0117] The above specific examples are only used to illustrate the invention to help understand the invention, and are not intended to limit the invention. Those skilled in the art may make several modifications and improvements based on the concept of the invention, which all belong to the protection scope of the invention.

[0118] Any matters not described in the present invention are applicable to the prior art.

Claims

1. A high-density lipoprotein cholesterol assay kit, characterized in that: Including reagent 1 and reagent 2, The components and concentrations of the reagent 1 are: The components and concentrations of the reagent 2 are:

2. The kit according to claim 1, characterized in that: The high molecular cationic polymer is a high polymer that can quickly aggregate and precipitate the precipitate produced by dextran sulfate and non-HDL lipoprotein from a dispersed state within 3 minutes.

3. The kit according to claim 1, characterized in that: The high molecular cationic polymer in the reagent 1 is at least one of an amine polymer, an imine polymer, and a polyquaternary ammonium salt; The amine polymer is at least one of polyamine and polyethyleneamine; the imine polymer is at least one of polyethyleneimine and polypropyleneimine; the polyquaternary ammonium salt is at least one of polyquaternary ammonium salt-5, polyquaternary ammonium salt-6, polyquaternary ammonium salt-7, and polyquaternary ammonium salt 39.

4. The kit according to claim 1, wherein: The high molecular cationic polymer in the reagent 1 is polyethyleneimine, and the molecular weight is not less than 3000.

5. The kit according to claim 1, wherein: The dextran sulfate in the reagent 1 is selected from at least one of: dextran sulfate-molecular weight 3000, dextran sulfate-molecular weight 20000, dextran sulfate-molecular weight 40000, and dextran sulfate-molecular weight 50000.

6. The kit according to claim 1, characterized in that: The buffers in the reagent 1 and the reagent 2 are independently selected from: phosphate buffer, 3-[NN-bis(2-hydroxyethyl)amino]-2-hydroxypropanesulfonic acid buffer, glycine buffer, 4-hydroxyethylpiperazineethanesulfonic acid buffer, 3-morpholinepropanesulfonic acid buffer; The inorganic salt in the reagent 1 is selected from one or more of sodium chloride, potassium chloride, magnesium chloride and calcium chloride; The surfactant 1 in the reagent 2 is selected from one or more of the emulsifiers AEO-3, AEO-7, and AEO-9; The preservatives in the reagent 1 and the reagent 2 are selected from one or more of sodium azide, PC300, gentamicin and sodium benzoate.

7. The kit according to claim 1, characterized in that: The concentrations of the components in the kit are: Reagent 1 is: Reagent 2 is: The pH value of the above buffer is 7.0-8.

0.

8. The kit according to any one of claims 1 to 7, characterized in that The upper limit of the anti-interference of the kit for chyle samples is increased to more than 2%; for lipid turbidity samples, the deviation of the multiple dilution back calculation of the kit is controlled within 10%.

9. A detection method of a high-density lipoprotein cholesterol determination kit, characterized in that: The process of the method is: after adding reagent 1 to the sample on the biochemical analyzer, incubate for 3 minutes, the high molecular cationic polymer adsorbs dextran sulfate DS and quickly aggregates and precipitates the dextran sulfate and non-HDL lipoprotein from a dispersed state within 3 minutes, so that the absorbance is restored to the initial level, that is, the absorbance level when no precipitation is generated, and then reagent 2 is added.

10. The detection method according to claim 9, characterized in that: The reagent 1 is a reagent containing dextran sulfate and a high molecular cationic polymer, which eliminates the interference of the fat turbidity sample by precipitation method; The reagent 2 is a reagent containing cholesterol oxidase, cholesterol esterase and 4-aminoantipyrine.

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