Hemolysis index quality control product as well as preparation method and application thereof
By developing a self-made hemolysis index quality control product, which utilizes mixed serum and red blood cells to destroy the sample and sodium azide as a preservative, the problems of high cost and poor stability of existing quality control products have been solved. This has enabled low-cost and efficient hemolysis sample detection management, ensuring the accuracy and consistency of test results.
Patent Information
- Application Number
- CN202511154695.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-18
- Publication Date
- 2025-10-28
AI Technical Summary
Existing hemolysis index quality control products are costly and have poor stability, and they are also insufficient in terms of monitoring and managing the detection of hemolyzed samples, which affects the accuracy and efficiency of clinical biochemical testing.
A self-made hemolysis index quality control was used. By mixing the remaining serum from the mixed test specimens with the destroyed red blood cells and the preservative sodium azide, quality control samples with different hemolysis indices were prepared to monitor the reaction and detection capabilities of IVD instruments.
To reduce costs, improve detection accuracy and efficiency, ensure the instrument's ability to detect hemolyzed samples, possess good stability and precision, reduce human error, and achieve automated detection and reliable results.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of medical testing and measurement technology, specifically to a hemolysis index quality control product, its preparation method, and its application. Background Technology
[0002] Internal quality control materials are a crucial material foundation for ensuring internal quality control work. Ensuring sample quality before testing is a vital aspect of routine and emergency laboratory testing. Clinical biochemistry testing, based on the premise of effective internal quality control using quality control materials, primarily utilizes theories and techniques from physics, chemistry, and biology to quantitatively analyze collected human body fluid samples, including serological and immunochemical examinations. Its role in clinical practice is mainly reflected in the following: Biochemical tests include diagnostically valuable items such as blood glucose and lipids. Accurate test results provide a scientific basis for clinical disease diagnosis, thus helping clinicians develop effective treatment plans and prevention and control measures. Biochemical tests such as liver function indicators, kidney function indicators, blood glucose, electrolytes, myocardial enzyme profiles, and C-reactive protein are involved in emergency testing, which is a crucial component of emergency treatment and resuscitation for critically ill patients. The timeliness and accuracy of emergency test results play a vital role in saving lives and alleviating suffering. Therefore, biochemistry testing plays a significant role in disease diagnosis, disease monitoring, efficacy observation, prognosis assessment, and disease prevention.
[0003] Numerous studies both domestically and internationally have demonstrated that the presence of high concentrations of interfering substances, such as hemoglobin, can reduce the quality of testing, interfering with results and leading to biased analysis. Hemolysis refers to the rupture of red blood cells, resulting in the release of hemoglobin from inside the cell to the extracellular space. The presence of hemolyzed specimens is often a major cause of delayed diagnosis, increased patient suffering, and prolonged reporting cycles. Historically, laboratory technicians have relied on visual inspection to judge specimen appearance, which is highly subjective and difficult to standardize and automate. In clinical laboratory diagnostics, the serum hemolysis index (HI) is a crucial indicator for assessing blood sample quality. Currently, domestic laboratory biochemical and immunoassay instruments can use the hemolysis index to evaluate specimen appearance quality. This involves measuring the absorbance at a specific wavelength to calculate the hemoglobin concentration in the serum or plasma, objectively reflecting the degree of hemolysis. The application of the serum hemolysis index function quantifies the criteria for judging specimen appearance, reducing the rate of rejected specimens due to hemolysis. Serum hemolysis index quality control products serve as the basis for quality assurance of this indicator.
[0004] Hemolysis index quality control products are used to monitor the reaction and detection capabilities of clinical biochemical testing instruments to hemolyzed samples. They typically contain a specific concentration of hemolytic substances to simulate samples with varying degrees of hemolysis. By comparing these samples with normal serum samples, the instrument's ability to detect hemolyzed samples can be assessed, thereby determining the instrument's accuracy and stability. Furthermore, quality control products can also be used to monitor the skill level of laboratory personnel and the quality of the laboratory environment. In summary, hemolysis index quality control products help ensure the accuracy and reliability of laboratory test results, improving laboratory efficiency and quality.
[0005] The research and application of quality control products have received widespread attention both domestically and internationally. Meanwhile, a series of relevant standards and regulations have been introduced in China, such as the "Clinical Laboratory Quality Management Standards," providing guidance and assurance for the research and application of quality control products. Existing hemolysis index quality control products on the market suffer from high costs and poor stability. Currently, research and preparation of quality control products in the field of biochemical testing conducted by medical and research institutions involve glycated hemoglobin protein controls, transfusion compatibility testing internal quality control products, etc., but research on the preparation of serum hemolysis index quality control products is scarce. Summary of the Invention
[0006] This invention aims to at least partially address one of the technical problems in related technologies. This research aims to develop a self-made hemolysis index quality control material designed to monitor the response and detection capabilities of in vitro diagnostic products (IVD), i.e., IVD instruments, to hemolyzed samples, to meet the needs of clinical laboratories. This quality control material can be used to assist laboratories in improving the detection of pre-analytical errors that affect clinical biochemical assays. The advantages of the hemolysis index quality control material provided by this invention are that it uses serum mixed with the remaining sample after testing, resulting in low cost and virtually no matrix effect. Its application and promotion in biochemical testing can not only save costs but also bring economic benefits.
[0007] Specifically, the present invention provides the following technical solution: A first aspect of the present invention provides a hemolysis index quality control product, comprising serum, preservatives, and hemolytic components; The serum, preservatives, and hemolytic components are mixed in different predetermined proportions to obtain quality control products with different hemolysis indices. The hemolyzed component is a blood sample containing destroyed red blood cells; The preservative is 0.05% sodium azide.
[0008] According to embodiments of the present invention, the hemolysis indices of the quality control products with different hemolysis indices are 20-50, 70-100, 100-200, 250-350, and 500-700, respectively.
[0009] According to specific embodiments of the present invention, the hemolysis indices of the quality control products with different hemolysis indices are 630, 300, 150, 75, and 37.5, respectively.
[0010] According to an embodiment of the present invention, the serum is a mixed serum of at least 10 subjects.
[0011] According to an embodiment of the present invention, the serum is a mixed serum from at least 10 subjects.
[0012] According to an embodiment of the present invention, the serum is a mixed serum of at least two subjects, and the hemolysing component is a blood sample with destroyed red blood cells from at least two subjects.
[0013] According to an embodiment of the present invention, the quality control samples with different hemolysis indices are obtained by the following method: Provide quality control products with a hemolysis index of 500-700, wherein the quality control products with a hemolysis index of 500-700 are blood samples with destroyed red blood cells; A quality control product with a hemolysis index of 250-350 is provided. The quality control product with a hemolysis index of 250-350 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 100-200 is provided. The quality control product with a hemolysis index of 100-200 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 70-100 is provided. The quality control product with a hemolysis index of 70-100 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 20-50 is provided, which is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide.
[0014] A second aspect of the present invention provides a method for preparing a hemolysis index quality control product, comprising: Blood was collected from at least two subjects. Serum was separated from a portion of the blood, inactivated, and then mixed to obtain mixed serum. A portion of the blood was frozen until the red blood cells were destroyed to obtain a blood sample with destroyed red blood cells. The mixed serum, preservatives, and hemolytic components are mixed in different predetermined proportions to obtain quality control products with different hemolysis indices. The hemolytic component is a blood sample containing destroyed red blood cells; The preservative is 0.05% sodium azide; The hemolysis indices of the quality control products with different hemolysis indices are 20-50, 70-100, 100-200, 250-350, and 500-700, respectively.
[0015] According to an embodiment of the present invention, the quality control samples with different hemolysis indices are obtained by the following method: Provide quality control products with a hemolysis index of 500-700, wherein the quality control products with a hemolysis index of 500-700 are blood samples with destroyed red blood cells; A quality control product with a hemolysis index of 250-350 is provided. The quality control product with a hemolysis index of 250-350 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 100-200 is provided. The quality control product with a hemolysis index of 100-200 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 70-100 is provided. The quality control product with a hemolysis index of 70-100 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 20-50 is provided, which is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide.
[0016] A third aspect of the present invention provides a kit comprising the hemolysis index control product described in the first aspect or the hemolysis index control product obtained according to the preparation method described in the second aspect.
[0017] The fourth aspect of the present invention provides the application of the hemolysis index quality control product described in the first aspect or the hemolysis index quality control product obtained according to the preparation method described in the second aspect in the diagnosis of hemolysis in non-disease conditions.
[0018] A fifth aspect of the present invention provides a method for determining the hemolysis index of a serum sample to be tested, comprising: The hemolysis index of the test sample was determined using an automated biochemical analyzer in conjunction with a hemolysis index quality control sample. The hemolysis index control product is the hemolysis index control product described in the first aspect or the hemolysis index control product obtained according to the preparation method described in the second aspect.
[0019] A sixth aspect of the present invention provides a method for detecting the reactivity of an instrument to a hemolyzed sample, comprising: The instrument is used to test the hemolysis index quality control sample. Based on the hemoglobin concentration in serum or plasma measured by the instrument, the hemolysis index is calculated. The hemolysis index quality control sample is the hemolysis index quality control sample described in the first aspect or the hemolysis index quality control sample obtained according to the preparation method described in the second aspect.
[0020] The beneficial effects achieved by this invention are at least as follows: The hemolysis index quality control provided by this invention can monitor the instrument's response and detection capability to hemolyzed samples, thus meeting the needs of clinical laboratories. Furthermore, the hemolysis index quality control is prepared in-house using serum, which offers advantages such as selecting residual samples after testing and mixing them with serum, resulting in low cost and virtually no matrix effect. This not only saves costs but also brings economic benefits.
[0021] The hemolysis index quality control provided by this invention remained essentially stable in average value after being stored at 4°C and -20°C for two weeks, indicating good stability under different storage conditions. Furthermore, the provided hemolysis index quality control exhibited good precision in terms of repeatability, intermediate precision, and reproducibility, with RSD values all less than 7%, meeting the requirements for clinical testing quality control. Detailed Implementation
[0022] The embodiments of the present invention are described in detail below. These embodiments are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention.
[0023] This study aims to develop a self-made hemolysis index quality control, designed to monitor the response and detection capabilities of clinical laboratory testing instruments to hemolyzed samples, thus meeting the needs of clinical laboratories. This quality control can assist laboratories in improving the detection of pre-analytical errors that affect clinical biochemical assays. The advantages of the self-made hemolysis index quality control include the use of residual serum from post-analysis samples, low cost, and minimal matrix effect. The preparation method involves adding fragmented red blood cells, stabilizers, preservatives, etc., to the mixed serum according to the hemolysis level (the threshold affecting each assay) to create a multi-concentration hemolysis index quality control, which not only saves costs but also brings economic benefits.
[0024] The preservative mentioned is 0.05% sodium azide, and this refers to the mass concentration. During the research, it was found that compared to other preservatives, sodium azide has broad-spectrum antibacterial activity. Sodium azide can effectively inhibit the growth of various microorganisms, including bacteria, fungi, and yeasts, thereby extending the shelf life of hemolyzed quality control products. It achieves its preservative effect by blocking enzymes in cellular respiration (such as cytochrome oxidase), inhibiting the metabolic activities of microorganisms; and it is stable under normal storage conditions. Furthermore, to ensure its effective preservative effect, a concentration of 0.05% was chosen.
[0025] The hemolysis index quality control samples provided can be repackaged, protected from light, and frozen.
[0026] The hemolysis index quality control provided by this invention has multiple uses, such as for monitoring and evaluating the quality of blood samples in clinical laboratory testing, especially in the specimen processing stage before analysis. Its main uses include: (1) Monitoring specimen quality: Hemolysis index quality control can help laboratories monitor the quality of blood specimen collection, delivery and processing, ensuring the integrity of the specimen and the accuracy of the test results. (2) Reducing errors and improving test quality: By using hemolysis index quality control, laboratories can effectively reduce erroneous test results caused by hemolysis, reduce specimen rejection rate and improve patient satisfaction. (3) Standardizing testing procedures: Hemolysis index quality control helps standardize laboratory testing procedures, ensuring the comparability of results between different laboratories, thereby improving the reliability and consistency of test results. (4) Automated testing and reporting: Combined with automated testing systems, hemolysis index quality control can realize automated testing and evaluation of hemolyzed specimens, reduce human error and improve work efficiency. (5) Interference detection and management: Hemolysis index quality control products can also be used to detect and manage possible interfering factors in the pre-analysis stage, such as hemolysis, jaundice and lipemia, thereby ensuring the accuracy of clinical biochemical test results.
[0027] In summary, hemolysis index quality control products play an important role in clinical laboratories. By monitoring and managing the quality of blood samples, they ensure the accuracy and reliability of test results, thereby providing a reliable basis for clinical diagnosis.
[0028] Accordingly, hemolysis index quality control products can be prepared into reagent kits.
[0029] The present invention also provides a method for detecting the reactivity of an instrument to a hemolyzed sample, comprising: The hemolysis index quality control sample is tested using the instrument, and the hemolysis index is calculated based on the hemoglobin concentration in serum or plasma measured by the instrument. The hemolysis index result is compared with the instrument's threshold to determine the instrument's responsiveness to hemolyzed samples.
[0030] The technical solution of the present invention will be described below through specific embodiments. It should be noted that these embodiments are only used to facilitate understanding by those skilled in the art and should not be considered as limiting the scope of protection of the present invention. Unless otherwise specified, the reagents used in the embodiments are all commercially available.
[0031] Example 1 Example 1: A hemolysis index control sample was prepared by the following method: Serum samples were collected from emergency patients over a month, and samples with lipemia, jaundice, or infectious agents were excluded. Hemoglobin solution that had been destroyed by freeze-thaw cycles was added to prepare serum hemolysis index quality control products with hemolysis indices of 600, 300, 150, 75, and 37.5, respectively. These products were then aliquoted into ampoules, with each ampoule containing 200 μL, and stored frozen at -20°C in the dark.
[0032] The quality control samples with different hemolysis indices can be prepared using the following method: (1) Prepare a high-concentration hemolytic stock solution for red blood cell destruction in advance. Its hemolysis index is 630, which can be used as a high-value stock solution or for future use. (2) Prepare a quality control product with a hemolysis index of 300. The preparation ratio is: 500ul mixed serum, 500ul high-concentration red blood cell destruction hemolysis stock solution (hemolysis index of 630), and then add 0.05% sodium azide preservative. (3) Prepare a quality control product with a hemolysis index of 150. The preparation ratio is: 750ul mixed serum, 250ul high-concentration red blood cell destruction hemolysis stock solution (hemolysis index of 630), and then add 0.05% sodium azide preservative. (4) Prepare a quality control product with a hemolysis index of 75. The preparation ratio is: 875ul mixed serum, 125ul high-concentration red blood cell destruction hemolysis stock solution (hemolysis index of 630), and then add 0.05% sodium azide preservative. (5) Prepare a quality control product with a hemolysis index of 37.5. The preparation ratio is: 937.5ul mixed serum, 62.5ul high-concentration red blood cell destruction hemolysis mother solution (hemolysis index is 630), and then add 0.05% sodium azide preservative.
[0033] Example 2 Taking Example 1 as an example, Example 2 verified the stability and precision of the hemolysis index quality control product prepared in Example 1: 1. Stability Quality control samples with different hemolysis indices were stored at 4℃ and -20℃ for two weeks at each temperature. Detection method: Samples were taken weekly, and the hemolysis index was measured using a fully automated biochemical analyzer. Data were recorded. Number of replicates: Each quality control sample with a hemolysis index was replicated three times, and the average value was taken.
[0034] The results are shown in Table 1 below: Table 1. Stability test results
[0035] As can be seen from the table, after the quality control samples with different hemolysis indices were stored at 4℃ and -20℃ for 2 weeks, the average value of the hemolysis index remained basically stable and the standard deviation was small, indicating that the quality control samples had good stability under both storage conditions.
[0036] 2. Precision Under identical conditions, perform 20 repeated measurements on the same quality control sample and calculate the relative standard deviation (RSD). Intermediate precision: Measure the same quality control sample by different operators at different time points and calculate the RSD. Reproducibility: Divide the quality control sample into multiple portions and perform measurements by different laboratories, then calculate the RSD.
[0037] Table 2 Precision Measurement Results
[0038] As can be seen from the table, all the quality control products for hemolysis index showed good precision in terms of repeatability, intermediate precision and reproducibility, with RSD values of less than 7%, which meets the requirements for quality control products for clinical testing.
[0039] Example 3 Example 3 utilizes the hemolysis index quality control sample to test the reactivity of an instrument for hemolyzed samples. The instrument used is the Orsonvir Vitros 5600 rapid biochemical immunoassay analyzer from the USA. The specific steps are as follows: (1) Set the hemolysis index (HI) measurement parameters according to the instrument manual and ensure that the instrument is in quality control mode.
[0040] (2) Testing the quality control sample: Add the hemolysis index quality control sample prepared in Example 1 to the instrument and perform the test according to the instrument's operating procedure. The instrument will automatically measure the hemoglobin concentration in serum or plasma and calculate the hemolysis index.
[0041] Results Analysis: After the test is completed, the instrument will display the hemolysis index result. If the result is within the manufacturer's allowable range (e.g., the precision for low and high values is 1.56% and 1.96%, respectively), it indicates that the instrument has a good ability to respond to hemolyzed samples.
[0042] Recording and Reporting: Record the test results in the Laboratory Information System (LIS) and generate a test report with annotations. For specimens with hemolysis index exceeding the interference threshold, an abnormality marker should be added to the report to alert the clinician.
[0043] The corresponding hemolysis index results are shown in Table 3 below:
[0044] All hemolysis index results were within the manufacturer's allowable precision range (1.56%~1.96%), indicating that VITROS 5600 has good detection capabilities for samples with different degrees of hemolysis.
[0045] By following the steps above, the instrument's ability to respond to hemolyzed samples can be effectively assessed, ensuring the accuracy and reliability of laboratory test results.
[0046] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.
Claims
1. A hemolysis index quality control product, characterized in that, Includes serum, preservatives, and hemolytic components; The serum, preservatives, and hemolytic components are mixed in different predetermined proportions to obtain quality control products with different hemolysis indices. The hemolytic component is a blood sample with destroyed red blood cells or a hemoglobin standard; The preservative is 0.05% sodium azide.
2. The hemolysis index quality control product according to claim 1, characterized in that, The hemolysis indices of the quality control products with different hemolysis indices are 20-50, 70-100, 100-200, 250-350, and 500-700, respectively.
3. The hemolysis index quality control product according to claim 1, characterized in that, The serum is a mixed serum from at least two subjects; Optionally, the serum is a pooled serum from at least 10 subjects; Optionally, the serum is a mixed serum from at least two subjects, and the hemolysing component is a blood sample with destroyed red blood cells from at least two subjects.
4. The hemolysis index quality control product according to claim 1, characterized in that, The quality control samples with different hemolysis indices were obtained by the following method: Provide quality control products with a hemolysis index of 500-700, wherein the quality control products with a hemolysis index of 500-700 are blood samples with destroyed red blood cells; A quality control product with a hemolysis index of 250-350 is provided. The quality control product with a hemolysis index of 250-350 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 100-200 is provided. The quality control product with a hemolysis index of 100-200 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 70-100 is provided. The quality control product with a hemolysis index of 70-100 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 20-50 is provided, which is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide.
5. A method for preparing a hemolysis index quality control product, characterized in that, include: Blood was collected from at least two subjects. Serum was separated from a portion of the blood, inactivated, and then mixed to obtain mixed serum. A portion of the blood was frozen until the red blood cells were destroyed to obtain a blood sample with destroyed red blood cells. The mixed serum, preservatives, and hemolytic components are mixed in different predetermined proportions to obtain quality control products with different hemolysis indices. The hemolytic component is a blood sample containing destroyed red blood cells; The preservative is 0.05% sodium azide; The hemolysis indices of the quality control products with different hemolysis indices are 20-50, 70-100, 100-200, 250-350, and 500-700, respectively.
6. The preparation method according to claim 5, characterized in that, The quality control samples with different hemolysis indices were obtained by the following method: Provide quality control products with a hemolysis index of 500-700, wherein the quality control products with a hemolysis index of 500-700 are blood samples with destroyed red blood cells; A quality control product with a hemolysis index of 250-350 is provided. The quality control product with a hemolysis index of 250-350 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 100-200 is provided. The quality control product with a hemolysis index of 100-200 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 70-100 is provided. The quality control product with a hemolysis index of 70-100 is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide. A quality control product with a hemolysis index of 20-50 is provided, which is obtained by mixing and compounding serum, a quality control product with a hemolysis index of 500-700, and 0.05% sodium azide.
7. A reagent kit, characterized in that, Includes the hemolysis index quality control product according to any one of claims 1 to 4 or the hemolysis index quality control product obtained by the preparation method according to claim 5 or 6.
8. The use of the hemolysis index control product according to any one of claims 1 to 4, or the hemolysis index control product obtained by the preparation method according to claim 5 or 6, in the diagnosis of hemolysis in non-disease conditions.
9. A method for determining the hemolysis index of a serum sample to be tested, characterized in that, include: The hemolysis index of the test sample was determined using an automated biochemical analyzer in conjunction with a hemolysis index quality control sample. The hemolysis index quality control product is the hemolysis index quality control product according to any one of claims 1 to 4 or the hemolysis index quality control product obtained by the preparation method according to claim 5 or 6.
10. A method for detecting the reactivity of an instrument to a hemolyzed sample, characterized in that, include: The hemolysis index quality control is tested using the instrument, and the hemolysis index is calculated based on the hemoglobin concentration in serum or plasma measured by the instrument. The hemolysis index quality control is the hemolysis index quality control according to any one of claims 1 to 4 or the hemolysis index quality control obtained by the preparation method according to claim 5 or 6. The hemolysis index result is compared with the instrument's threshold to determine the instrument's responsiveness to hemolyzed samples.