Preparation method of quality control product for conventional blood coagulation items
Through the method of preparing conventional coagulation quality control products, the problem of expensive commercial quality control products is solved, and uniform and stable conventional coagulation project detection is achieved, reducing costs and improving detection capabilities.
Patent Information
- Application Number
- CN202510585494.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-08
- Publication Date
- 2025-08-08
AI Technical Summary
In the prior art, most of the coagulation quality control substances used in clinical laboratories are commercial quality control substances, which have matrix effects and are expensive, which increases the cost of laboratory reagents. The quality assurance of homemade quality control substances is of great significance.
Physical and chemical methods are used to process mixed plasma for multiple people, prepare conventional coagulation quality control products at different concentration levels, and store them in ultra-low temperature refrigerators, including centrifugation, mixing, dilution and aliquoting steps to ensure the uniformity and stability of the product.
The prepared conventional coagulation items have uniformity and stability, and can replace commercial products for quality control, reduce testing costs, and improve testing capabilities.
Smart Images
Figure CN120446507A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of clinical coagulation test preparations, and in particular to a method for preparing a quality control product for routine coagulation items. Background Art
[0002] Routine coagulation test items are suitable for screening, diagnosis, treatment effect evaluation, and differential diagnosis of thrombotic and hemorrhagic diseases. There are many reagents and instruments used for routine coagulation test items. Different reagents, test instruments, and test principles vary. It is very important to use routine coagulation test item quality control products to perform indoor and inter-laboratory quality control of routine coagulation test items. According to the relevant industry standards for laboratory medicine, the indoor quality control of coagulation items includes at least two levels of quality control products, namely normal level (test results within the reference range) and abnormal level (test results outside the reference range) quality control products.
[0003] Currently, most coagulation quality control materials used in clinical laboratories are commercial quality control materials, which have matrix effects and are expensive, increasing the cost of laboratory reagents. Therefore, laboratory-made quality control materials are of great significance to the quality assurance of routine coagulation projects. Summary of the Invention
[0004] In order to solve the above technical problems, the present invention designs a method for preparing a quality control product for routine coagulation items.
[0005] In order to achieve the above technical effects, the present invention is implemented by the following technical solutions: a method for preparing a quality control product for routine coagulation items, characterized in that it comprises the following steps:
[0006] S1. Blood collected from a human vein was mixed with a sodium citrate anticoagulant at a concentration of 0.109 mol / L at a volume ratio of 9:1. The mixture was centrifuged at a temperature of 18°C to 25°C and a centrifugal force of 1500-1700 g for 15-20 minutes. The plasma sample was tested for routine coagulation tests using a fully automatic coagulation analyzer.
[0007] S2. Screen the results of routine coagulation tests, separate and aspirate the upper plasma of samples whose results are within the reference range, and mix the upper plasma of multiple samples to obtain mixed plasma;
[0008] S3. Centrifuging the obtained normal mixed plasma for multiple people at a temperature of 18° C. to 25° C. and a centrifugal force of 1500 to 1700 g for 15 to 20 minutes to obtain treated mixed plasma for multiple people;
[0009] S4. Prepare a quality control sample for abnormal coagulation levels by adding 200 ml of coagulation analysis diluent to every 100 ml of the processed multi-person mixed plasma.
[0010] S5. Perform quality control of routine coagulation test items, divide the package, and store in an ultra-low temperature refrigerator at -75℃ to -85℃.
[0011] Furthermore, in S1, the blood collected from the human vein is sampled from multiple healthy donors and patient samples.
[0012] Furthermore, in S1, the routine coagulation items include prothrombin time PT, activated partial thromboplastin time APTT, thrombin time TT, fibrinogen FIB and antithrombin III ATIII.
[0013] Furthermore, in S2, the reference interval is the reference interval for each routine coagulation item, specifically as follows:
[0014] PT: 9.4-12.5 seconds; APTT: 25.1-36.5 seconds; FIB: 2.38-4.98 seconds; TT: 10.3-16.6 seconds; ATⅢ: 83-128 seconds;
[0015] Furthermore, the diluent for coagulation analysis is prepared as follows:
[0016] 1) Weigh 4.42 g of barbital sodium, then add water to the barbital sodium to completely dissolve it and dilute to 400 ml to obtain a stock solution;
[0017] 2) adding 0.1% sodium azide to the original solution to obtain an initial solution;
[0018] 3) Use 2 mol / L hydrochloric acid solution to adjust the pH value of the initial solution to 7.35, and then filter to obtain a diluted solution.
[0019] The beneficial effects of the present invention are:
[0020] The quality control product for routine coagulation items prepared by the present invention has good product uniformity, stability and stability after frozen plasma rethawing. It can replace commercial products for quality control of routine coagulation item detection, which is beneficial to reducing detection costs and improving the laboratory's ability to detect routine coagulation items. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.
[0022] Figure 1 It is a flow chart for preparing the coagulation factor quality control product of the present invention. DETAILED DESCRIPTION
[0023] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.
[0024] Example 1
[0025] Prepare quality control products for routine coagulation tests with normal results;
[0026] The preparation method of quality control products for routine coagulation tests includes the following steps:
[0027] 1) Blood collected from multiple healthy donors was mixed with 0.109 mol / L sodium citrate anticoagulant at a volume ratio of 9:1. The samples were centrifuged at 1500 g for 15 minutes at a temperature of 18°C to 25°C. The plasma samples were then tested for routine coagulation tests using an automated coagulation analyzer. Routine tests included prothrombin time (PT), apoptotic thrombosis (APTT), thrombotic time (TT), fibrillation in the urethra (FIB), and AT III.
[0028] 2) Screen the coagulation test results, separate and aspirate the upper plasma of samples with results within the reference range, and mix the upper plasma of multiple samples to obtain multiple normal mixed plasma;
[0029] 3) centrifuging the normal plasma pool of multiple individuals at a temperature of 18° C. to 25° C. and a centrifugal force of 1500 g for 15 minutes to obtain processed plasma pool of multiple individuals;
[0030] 4) Prepare a coagulation control sample for abnormal concentration levels by adding 200 ml of coagulation analysis diluent (saline solution and less than 0.1% sodium azide) to every 100 ml of the processed multi-person pooled plasma.
[0031] 5) The prepared coagulation routine items are quality controlled, packaged, and stored in an ultra-low temperature freezer at -80°C.
[0032] Example 2
[0033] Prepare quality control products for routine coagulation tests with abnormal results;
[0034] Blood samples from patients were mixed with 0.109 mol / L sodium citrate anticoagulant at a volume ratio of 9:1 and centrifuged at 1500 g for 15 minutes at a temperature of 18°C to 25°C. The plasma samples were then tested for routine coagulation tests using an automatic coagulation analyzer. Routine tests included PT, APTT, TT, FIB, and ATIII.
[0035] 2) Screen the coagulation test results, separate and aspirate the upper plasma of samples with results within the reference range, and mix the upper plasma of multiple samples to obtain multiple abnormal mixed plasma;
[0036] 3) centrifuging the abnormal mixed plasma from multiple individuals at a temperature of 18° C. to 25° C. and a centrifugal force of 1500 g for 15 minutes to obtain processed mixed plasma from multiple individuals;
[0037] 4) Prepare a coagulation control sample for abnormal concentration levels by adding 200 ml of coagulation analysis diluent (saline solution and less than 0.1% sodium azide) to every 100 ml of the processed multi-person pooled plasma.
[0038] 5) The prepared coagulation routine items are quality controlled, packaged, and stored in an ultra-low temperature freezer at -80°C.
[0039] Example 3
[0040] The products obtained in Example 1 and Example 2 were tested as follows:
[0041] The IL-TOP 750 fully automatic coagulation analyzer and the supporting coagulation routine test reagents were used to test the coagulation routine items of the unpackaged mixed plasma in Example 1 and Example 2 (see
[0042] Table 1);
[0043]
[0044] Table 1 Coagulation test results of routine coagulation control products
[0045] As can be seen from the above table, the product obtained by this preparation method can meet the requirements of indoor quality control of coagulation projects and can obtain quality control products with normal and abnormal results.
[0046] Example 4
[0047] The quality control products after packaging in Example 1 and Example 2 were subjected to uniformity testing to ensure that the variation between units was small, as follows:
[0048] Homogeneity verification was performed using a one-way ANOVA. Ten subpackaged control samples were randomly selected and numbered 1 to 10. Each packaging unit was measured three times in the following order: 1, 3, 5, 7, 9, 2, 4, 6, 8, 10; 10, 9, 8, 7, 6, 5, 4, 3, 2, 1; and 2, 4, 6, 8, 10, 1, 3, 5, 7, 9. The measurement results for each control sample were recorded for subsequent calculations, including the following indicators:
[0049] To check the uniformity of the samples, i = 10, m = 10 samples were taken and each sample was tested j = 3, n = 3 times under repeated conditions.
[0050] The average value of each sample tested:
[0051] The total average value of all sample tests:
[0052] Total number of tests:
[0053] Between-sample sum of squares: mean square
[0054] Within-sample sum of squares: mean square
[0055] Degrees of freedom: f1 = m-1 = 10-1 = 9
[0056] f2=Nm=30-10=20
[0057] Statistics:
[0058] Interpretation of results: If F < the critical value F with the degrees of freedom (f1, f2) and a given significance level α (usually α = 0.05), α (f1, f2) = 2.39, indicating that there is no significant difference within and between samples, and the samples are uniform.
[0059] Substitute the subpackaged quality control products obtained in Example 1 and Example 2 into the above steps, and the results are shown in Table 2;
[0060] Table 2 Homogeneity verification results of quality control products for routine coagulation items
[0061]
[0062] As can be seen from the above table, there is no significant difference within or between samples of the products obtained by this preparation method, and the samples are uniform.
[0063] Example 5
[0064] Stability: Verify the re-dissolution stability and shelf life stability.
[0065] Reconstitution stability verification method: Take out the homemade quality control product stored in a -80℃ refrigerator for less than 6 months, place it in a 37℃ water bath for reconstitution for 5 minutes, test immediately, place it at room temperature for 1 hour and test again, repeat the measurement 3 times immediately, and repeat the measurement 3 times after 1 hour. Take the average value as X0 and X, calculate the relative deviation B (see Table 3 for requirements), calculation formula:
[0066] Shelf life stability verification method: comparison of the average value of a series of measurements with the certified value / reference value
[0067] Calculate the t value as follows:
[0068]
[0069] The average value of n measurements;
[0070] M: standard value / reference value;
[0071] n: number of measurements;
[0072] S: standard deviation of n measurement results.
[0073] Note: To ensure the accuracy of the mean and standard deviation, n≥6.
[0074] If t < significance level ɑ (usually ɑ = 0.05) and the degrees of freedom are n-1, the critical value t ɑ(n-1) , there is no significant difference between the mean value and the standard value / reference value.
[0075] Table 3 Stability verification results of quality control products for routine coagulation items
[0076]
[0077]
[0078] In summary, the quality control products for routine coagulation items prepared by this preparation method have good product uniformity, stability and stability after rethawing of frozen plasma. They can replace commercial products for quality control of routine coagulation item detection, which is beneficial to reduce detection costs and improve the laboratory's ability to detect routine coagulation items.
Claims
1. A method for preparing a quality control product for routine coagulation tests, characterized in that: The following steps are involved: S1. Blood collected from a human vein was mixed with 0.109 mol / L sodium citrate anticoagulant at a volume ratio of 9:
1. The mixture was centrifuged at a temperature of 18°C to 25°C and a centrifugal force of 1500-1700 g for 15-20 minutes. The coagulation parameters of the plasma sample were tested using an automatic coagulation analyzer. S2. Screen the results of routine coagulation tests, separate and aspirate the upper plasma of samples whose results are within the reference range, and mix the upper plasma of multiple samples to obtain mixed plasma; S3. Centrifuging the obtained mixed plasma at a temperature of 18° C. to 25° C. and a centrifugal force of 1500 to 1700 g for 15 to 20 minutes to obtain processed mixed plasma. S4. Prepare a quality control sample for abnormal coagulation levels by adding 200 ml of coagulation analysis diluent to every 100 ml of the processed multi-person mixed plasma. S5. Perform quality control of routine coagulation test items, divide the package, and store in an ultra-low temperature refrigerator at -75℃ to -85℃.
2. The method for preparing a quality control product for routine coagulation items according to claim 1, characterized in that: In S1, the blood collected from the human vein is sampled from multiple healthy donors and patient samples.
3. The method for preparing a quality control product for routine coagulation items according to claim 1, characterized in that: In S1, the routine coagulation items include prothrombin time PT, activated partial thromboplastin time APTT, thrombin time TT, fibrinogen FIB and antithrombin III ATIII.
4. The method for preparing a quality control product for routine coagulation items according to claim 1, characterized in that: In S2, the reference intervals are the reference intervals for each routine coagulation item, specifically as follows: PT: 9.4-12.5 seconds; APTT: 25.1-36.5 seconds; FIB: 2.38-4.98 seconds; TT: 10.3-16.6 seconds; ATⅢ: 83-128 seconds.
5. The method for preparing a quality control product for routine coagulation items according to claim 1, characterized in that: The coagulation analysis diluent was prepared as follows: 1) Weigh 4.42 g of barbital sodium, then add water to the barbital sodium to completely dissolve it and dilute to 400 ml to obtain a stock solution; 2) adding 0.1% sodium azide to the original solution to obtain an initial solution; 3) Use 2 mol / L hydrochloric acid solution to adjust the pH value of the initial solution to 7.35, and then filter to obtain a diluted solution.