Blood coagulation seven-item composite quality control product and preparation method thereof

Abstract

The invention belongs to the technical field of blood coagulation detection, and particularly relates to a blood coagulation seven-item composite quality control product and a preparation method thereof.The quality control product is divided into a level 1 and a level 2, and seven indexes include APTT, PT, TT, FIB, AT-III, D-Dimer and FDP. The technology is stable and reliable; any index of APTT, PT, TT, FIB, AT-III, D-D and FDP of the level 1 quality control product can be adjusted to any target value in a normal range, any index of seven indexes of the level 2 quality control product can be adjusted to any target value in an abnormal value range, and the target value requirements of various blood coagulation detection systems can be met; the freeze-dried product is good in stability after being redissolved; and the quality control product is excellent in uniformity.

Description

Technical Field

[0001] The invention belongs to the technical field of coagulation detection, and in particular relates to a seven-item composite quality control product for coagulation and a preparation method thereof. Background Art

[0002] The seven-item composite quality control product for coagulation is a substance with known content and clear characteristics in the same matrix as the actual specimen. It is mainly used in the coagulation detection process for precision testing of detection equipment and reagents and daily inter-laboratory and internal quality control in the laboratory. It has strict requirements on product uniformity, reconstitution stability, etc.

[0003] The seven coagulation tests refer to the seven tests of APTT, PT, TT, FIB, AT-III, FDP and D-Dimer carried out on the clinical samples of patients. These seven tests cover the three major related systems of coagulation, anticoagulation and fibrinolysis, and have great clinical significance for the diagnosis, monitoring and treatment of coagulation, anticoagulation, thrombolysis and other diseases of patients.

[0004] APTT test can reflect: 1. Deficiency of coagulation factors in the intrinsic coagulation pathway (such as factor VIII (hemophilia A), factor IX (hemophilia B), factor XI and factor XII, prothrombin, fibrinogen and factor V, factor X); 2. Increase of anticoagulants in the blood (lupus anticoagulant, coagulation factor inhibitors or heparin); 3. And some immune diseases (liver disease, obstructive jaundice, neonatal hemorrhagic disease and large-scale transfusion of stored blood, etc.), which has important clinical significance.

[0005] PT is an indicator that reflects the activity of coagulation factors I, II, V, VII, and X in plasma and evaluates the coagulation function in vivo. It is the basis for clinical adjustment of anticoagulant dosage. PT detection is also a screening test to check whether the body's exogenous coagulation system function is impaired. Specific phenomena that can cause PT prolongation include: 1. Congenital coagulation factor deficiency (such as prothrombin (factor II), factor V, factor VII, factor X, and fibrinogen (factor I)); 2. Acquired coagulation factor deficiency (such as secondary / primary hyperfibrinolysis, severe liver disease, etc.); 3. Use of heparin; 4. The presence of antibodies in the blood circulation (such as antibodies to prothrombin, factor V, factor VII, factor X, and fibrinogen). Specific phenomena that can cause PT shortening include: 1. Oral contraceptives; 2. Congenital factor V excess; 3. Embolic diseases; 4. Hypercoagulable state, etc.

[0006] TT is the time it takes for standardized thrombin to coagulate in plasma. Since fibrinogen degradation products (FDP) can prolong TT, it can be used as a screening test for the fibrinolytic system to reflect the conversion of fibrinogen to fibrin in the common coagulation pathway. Clinically, the phenomena that can lead to prolonged TT include: increased heparin or the presence of heparin-like anticoagulants, increased fibrinogen degradation products, disseminated intravascular coagulation (DIC), and hypo(an)fibrinogenemia.

[0007] FIB is the precursor of fibrin, an important protein involved in the coagulation and hemostasis process, and its concentration changes have an important impact on coagulation function. Clinically, increased FIB is an important risk factor for various thrombotic diseases, and is seen in diabetes, acute myocardial infarction, rheumatic diseases, acute glomerulonephritis, nephrotic syndrome, multiple myeloma, burns, shock, after major surgery, malignant tumors and prethrombotic states, the elderly, etc., while decreased FIB is common in DIC, primary fibrinolysis, severe hepatitis and cirrhosis, hypofibrinogenemia; it can also be seen in the use of fiber-lowering drugs (antithrombin, defibrostase) and thrombolytic therapy.

[0008] AT-III is an important anticoagulant in the body, which can inhibit the activity of thrombin and coagulation factors IXa, Xa, XIa, XIIa, etc., thereby regulating the coagulation process and maintaining the balance between coagulation and anticoagulation. Clinically, AT-III deficiency or reduced activity can lead to a hypercoagulable state of the blood, which is prone to thrombosis. It is common in diseases such as congenital AT-III deficiency, DIC, liver disease, and nephrotic syndrome. In addition, in terms of anticoagulant therapy monitoring, the effects of certain anticoagulant drugs (such as heparin) depend on AT-III. If the AT-III level is too low, the anticoagulant effect of heparin may not be fully exerted. At this time, AT-III needs to be supplemented or the anticoagulant therapy regimen needs to be adjusted.

[0009] FDP is a degradation product produced after the decomposition of fibrinogen and fibrin. Its content can reflect the intensity of fibrinolytic activity in the body.

[0010] D-Dimer is a soluble fibrin degradation product and can be used as an important molecular marker reflecting the hypercoagulable state of plasma and the activation of the fibrinolytic system in vivo.

[0011] Therefore, in the diagnosis, detection and treatment of blood system diseases, especially in recent years when thrombosis-related diseases have received more and more attention, the seven coagulation tests will play an important role in the monitoring, prevention and treatment of diseases such as thrombosis.

[0012] In order to improve clinical monitoring efficiency and reduce monitoring costs, more and more coagulation test equipment manufacturers and reagents focus on the development of integrated coagulation test systems that include the above seven coagulation items. As a result, there is a demand for quality control products for the seven coagulation tests. In the past, coagulation quality control products were mainly used to test single items or the first 4-5 coagulation indicators, and the development of composite quality control products for the seven coagulation items was relatively rare.

[0013] The specification of Chinese patent CN117741166A records a multi-item composite coagulation quality control product and its preparation method. The method uses human plasma and human serum to add coagulation factors V, VII, VIII, IX, X, XI, XII, AT-III, DD, FDP and fibrinogen to prepare a composite quality control product. The quality control product of the invention can be used for clinical PT, INR, APTT, TT, FIB, AT-III, DD and FDP quality control at the same time.

[0014] The specification of Chinese patent CN118425538A records seven composite quality control products for abnormal coagulation and their preparation methods. Rabbit plasma is used as matrix plasma, and hirudin is mainly used as an anticoagulant to adjust the relevant coagulation time items to prepare abnormal value quality control products, which can be used for PT, APTT, TT, FIB, AT-Ⅲ, D-Dimer, and FDP items at the same time. Rabbit plasma is used entirely as matrix plasma in this patent, which may be different from the human blood matrix environment during actual detection. If conditions permit, it is most ideal to use human plasma as the main component. The addition of hirudin from non-human materials will also increase the matrix effect. And this method is only used to prepare abnormal value quality control products, and normal quality control products are not yet included, so this method has great limitations.

[0015] The coagulation process is a process in which multiple factors (coagulation factors and other conditions) work together and restrict each other, which is ultimately reflected in PT, APTT, TT, FIB, AT-Ⅲ, D-Dimer, FDP and other items. It is difficult to debug a single item without changing other items. In addition, due to the differences in biological raw materials, quality control products may have large batch differences, and even the target value cannot be successfully debugged.

[0016] The specification of Chinese patent CN117741166A records the use of serum as one of the main components of the mixed matrix. Since serum involves the activation of a series of coagulation factors during its formation, these activated coagulation factors will bring uncertainty to the long-term stability of multiple coagulation quality control products, so multiple quality control product matrices should try to avoid or mix human serum. And this patent increases the cost required for coagulation factor purification by adding a preparation method for coagulation factors.

[0017] Therefore, there is an urgent need for a method for preparing a seven-item composite quality control product for coagulation, which can be adapted to any human plasma raw material, and the quality control product prepared by this method is long-term stable, and the content or measured value of each item can be arbitrarily adjusted within the normal range or the abnormal value range to meet the needs of multi-system testing or quality control requirements. Summary of the invention

[0018] In order to solve the above technical problems, the present invention provides a seven-item composite quality control product for coagulation and a preparation method thereof, which adopts human plasma as the matrix plasma to avoid the matrix effect, and has a reliable preparation process and strong operability. The prepared quality control product can meet the requirements of a wide range of target values ​​and accurately match any target value required for quality control, and at the same time has wide reagent and instrument applicability and long-term stability.

[0019] A method for preparing a seven-item composite quality control product for blood coagulation in the present invention that solves the above technical problems comprises the following steps: (1) Obtain sodium citrate anticoagulated human mixed plasma and sodium citrate anticoagulated animal plasma PP for use; Some coagulation factors in plasma can easily lose their activity, so the fresher the raw plasma, the better.

[0020] The human mixed plasma used is normal human plasma, but in practice, due to factors such as freezing and storage, the coagulation activity is attenuated, and the APTT and PT values ​​are prolonged. PP, the activity of coagulation factors is stronger than that of humans, and has shorter APTT and PT values ​​than humans. The role of animal plasma (PP) is to adjust the APTT and PT of quality control products; plasma that has lost some activity needs more PP to adjust to the APTT and PT values ​​corresponding to fresh human mixed plasma.

[0021] PP, in practical applications, can be screened by detection, where the APTT measurement value is in the range of 14 to 18 seconds.

[0022] (2) A portion of human mixed plasma and PP were adsorbed by aluminum hydroxide (Al(OH)3) colloid, and the supernatant was collected by centrifugation to obtain adsorbed human mixed plasma and adsorbed PP; In the present invention, both level 1 and level 2 need to be adsorbed by aluminum hydroxide colloid. The coagulation detection system includes a coagulation detection instrument and applicable APTT, PT, TT, FIB, AT-III, D-Dimer and FDP detection reagents. When detecting the same plasma sample, different systems often have inconsistent measurement values; the manufacturer of the detection system has its own target value for each indicator of the quality control product. The method of the present invention is to meet any detection system and adjust the detection values ​​of the seven indicators of the quality control product to the target value.

[0023] (3) Buffering stabilizing protective agents were added to another part of human mixed plasma and PP, as well as adsorbed human mixed plasma and adsorbed PP, to obtain the corresponding R0, PP0, R1 and PP1.

[0024] (4) Divide R0 and R1 into two parts: add PP0, the first part R1 and FIB adjustment solution to the first part R0, adjust the FIB, APTT, PT and TT values ​​to be consistent with fresh normal human mixed plasma, and obtain level 1 matrix solution 1, in which the target values ​​of APTT are 21~45sec, PT is 10~15sec, TT is 10~20sec, and FIB is 2~4mg / L; Add PP1, the second part R0, the diluent and the FIB adjustment solution to the second part R1, adjust the APTT, PT, TT, FIB and AT-Ⅲ values ​​to the target values, and obtain the level 2 matrix solution 1, wherein the target values ​​APTT is 45~150sec, PT is 15~74sec, TT is 20~50sec, FIB is 0.6~2mg / L, and AT-Ⅲ is 25~80%.

[0025] (5) Add DD stock solution and FDP stock solution to Level 1 Matrix Solution 1, and adjust the concentrations of DD and FDP in Level 1 Matrix Solution 1 to the target values ​​of Level 1 Matrix Solution 2, DD is 0.1-0.9 μg / L (FEU), and FDP is 2.2-5.5 μg / L; Then add AT-Ⅲ concentrate to level 1 matrix solution 2, and adjust the AT-Ⅲ value to be consistent with normal human mixed plasma, thus obtaining the seven-item coagulation composite quality control product level 1, in which the AT-Ⅲ target value is 80~120%; Add DD stock solution and FDP stock solution to level 2 matrix solution 1, adjust the concentrations of DD and FDP in level 2 matrix solution 1 to the target values ​​to obtain the coagulation seven-item composite quality control product level 2, where the target values ​​for DD are 0.9~30μg / L, and for FDP are 5.5~55μg / L.

[0026] (6) The two-level activated coagulation test reagent quality control product is obtained by subpackaging and freeze-drying.

[0027] Level 1 is the normal value control product; Level 2 is the abnormal value control product.

[0028] In the present invention, there is no need to adjust the pH, because the buffering stabilizing protective agent has already completed the function of adjusting the pH.

[0029] In the optimized solution, the amount (V / V) of aluminum hydroxide colloid in step (2) is 1% to 10%.

[0030] In step (1), PP is an animal plasma from cattle, pigs, sheep, and horses; and the centrifugation parameters are 4°C, 4000 rpm, and centrifugation for 10-50 min.

[0031] Furthermore, the buffer stabilizing protective agent in step (3) is one or more of trehalose, glycine, HEPES, sodium chloride, trisodium citrate, Tris and Proclin300.

[0032] Furthermore, the percentage concentration range of glycine is 0~160 g / L, the percentage concentration range of HEPES is 100~250g / L, the percentage concentration range of trehalose is 100~500g / L, the percentage concentration range of sodium chloride is 0~10g / L, the percentage concentration range of trisodium citrate is 0~5g / L, the percentage concentration range of Tris is 0~5g / L, and the concentration range (V / V) of Proclin300 is 1 / 1000 to 10.

[0033] The amount of buffer stabilizing agent added in step (3) (V / V) is 3% to 7%, i.e., 3-7 ml is added to 100 ml.

[0034] In step (4), the FIB regulating solution is human plasma concentrate, and its addition amount is 0-50% of the total volume of R0 and R1; the addition amount of PP0 or PP1 accounts for 0-20% of the total volume; the diluent accounts for 0-60% of the total volume; and R0 accounts for 0-20% of the total volume.

[0035] The diluent in the present invention is one or more of D-trehalose, sodium chloride, HEPES FREE ACID, dihydrate and trisodium citrate.

[0036] Furthermore, the percentage concentration range of D-trehalose is 0-100 g / L, the percentage concentration range of sodium chloride is 0-50 g / L; the percentage concentration range of HEPES FREE ACID is 0-30 g / L; and the percentage concentration range of trisodium citrate dihydrate is 0-5 g / L.

[0037] Furthermore, the ranges required for Level 1 of the seven-item composite quality control product for coagulation are a prothrombin time of 10 to 15 seconds, an activated partial thromboplastin time of 21 to 45 seconds, a fibrinogen concentration of 2 to 4 g / L, an antithrombin III activity of 80 to 120%, a D-dimer concentration of 0.1 to 0.9 μg / mL (FEU), a fibrin degradation product concentration of 2.5 to 5.5 μg / mL, and a thrombin time of 10 to 20 seconds.

[0038] The ranges required for the level 2 of the seven-item composite quality control product for coagulation are as follows: prothrombin time 15~74s, activated partial thromboplastin time 45~150s, fibrinogen concentration 0.6~2 g / L, antithrombin III activity 5~80%, D-dimer concentration 0.9~30 μg / mL (FEU), fibrin degradation product concentration 5.5~55 μg / mL, and thrombin time 20~50s.

[0039] The preparation method of the seven-item composite quality control product of coagulation can be adjusted to obtain level 1 quality control products (any index of APTT, PT, TT, FIB, AT-III, DD and FDP can be adjusted to any target value within the normal range) and level 2 quality control products (any index of the seven indexes can be adjusted to any target value within the abnormal value range).

[0040] The seven-item composite quality control product of coagulation prepared by any one of the methods described above in the present invention is a seven-item composite quality control product of two-level activated coagulation.

[0041] Furthermore, the required ranges of the medium indicators of the seven-item composite quality control product level 1 of coagulation are: APTT 25~31s, PT10.8~13s, TT 14~18s, Fib 2.2~3.2g / L, AT-Ⅲ 86~105%, DD 0.3~0.5 ug / ml (FEU), and FDP2.5~5ug / ml.

[0042] The required ranges of the indicators in the level 2 of the seven-item composite quality control product for coagulation are: APTT 48~65s, PT 28~36s, TT28~50s, Fib 1.2~1.7g / L, AT-Ⅲ 30~50%, DD 2.0~3.0ug / ml (FEU), and FDP 15~20ug / ml.

[0043] Furthermore, the fixed components and volume contents of the seven-item composite quality control product level 1 of coagulation are: R0 36~81%, R1 0~45%, FIB adjustment solution 0~8.2%, PP0 0~8.2%, FDP 0~0.73%, DD 0~0.73%, and AT III 0~0.3%.

[0044] The fixed components and volume contents of the seven-item composite quality control product level 2 for coagulation are: R1 14-37%, diluent 37-59%, FIB adjustment solution 0-3%, R0 0-11%, PP1 0-15%, FDP 0-0.73%, and DD 0-0.73%.

[0045] The content of each component in the present invention has a great influence on the result, and each step is used to facilitate the adjustment of the target value of each item index.

[0046] The seven-item coagulation composite quality control product and the preparation method thereof of the present invention are based on many years of experience in the research and development and production of coagulation series quality control products, and have undergone theoretical and process reliability evaluation, pilot verification and mass production tests.

[0047] The quality control product prepared by the method of the present invention has been tested by various coagulation systems and has excellent performance in terms of product uniformity, reconstitution stability, etc.; for FIB, FDP and DD tests that require quantitative detection values, good precision and target value accuracy are maintained within the detection range; APTT, PT, TT and AT III level tests show good precision and uniformity. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 Schematic diagram of the preparation process of the seven-item composite quality control product level 1 of coagulation in the present invention; Figure 2 Schematic diagram of the preparation process of the seven-item composite quality control product level 2 of coagulation in the present invention. DETAILED DESCRIPTION

[0049] The present invention is further described below in conjunction with specific embodiments: The present invention specifically relates to a preparation method of two levels of seven-item composite quality control products for coagulation, a seven-item composite quality control product LI for coagulation and a seven-item composite quality control product LII for coagulation. The seven coagulation items are partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), FIB (fibrinogen), antithrombin III (AT-III), fibrinolytic substance (FDP) and D-dimer (D-Dimer) related to thrombolysis.

[0050] The seven-item composite quality control product of coagulation in the present invention is a quality control product divided into level 1 and level 2, and the seven indicators include APTT, PT, TT, FIB, AT-III, D-Dimer and FDP. The preparation method of the seven-item composite quality control product of coagulation comprises the following steps: (1) Obtain sodium citrate anticoagulated human mixed plasma and sodium citrate anticoagulated animal plasma for later use; (2) Part of the human mixed plasma and PP are adsorbed by aluminum hydroxide (Al(OH)3) colloid, respectively, and the supernatant is collected by centrifugation to obtain adsorbed human mixed plasma and adsorbed PP; the amount of aluminum hydroxide colloid (V / V) is 1% to 10%; PP is an animal plasma from cattle, pigs, sheep, and horses; the centrifugation parameters are 4°C, 4000 rpm, and centrifugation for 10-50 min.

[0051] (3) Add buffering and stabilizing protective agents to another part of human mixed plasma and PP, as well as adsorbed human mixed plasma and adsorbed PP, to obtain the corresponding R0, PP0, R1 and PP1; The buffer stabilizing protective agent is one or more of trehalose, glycine, HEPES, sodium chloride, trisodium citrate, Tris and Proclin 300. The amount of buffer stabilizing protective agent added (V / V) is 3%~7%. The volume ratio is 3-7ml in 100ml.

[0052] The percentage concentration range of glycine is 0~160 g / L, the percentage concentration range of HEPES is 100~250g / L, the percentage concentration range of trehalose is 100~500g / L, the percentage concentration range of sodium chloride is 0~10g / L, the percentage concentration range of trisodium citrate is 0~5g / L, the percentage concentration range of Tris is 0~5g / L, and the concentration range of Proclin300 (V / V) is 1 / 1000 to 10.

[0053] (4) Divide R0 and R1 into two parts: add PP0, the first part R1 and FIB adjustment solution to the first part R0, adjust the FIB, APTT, PT and TT values ​​to be consistent with fresh normal human mixed plasma, and obtain level 1 matrix solution 1, in which the target values ​​of APTT are 21~45sec, PT is 10~15sec, TT is 10~20sec, and FIB is 2~4mg / L; Add PP1, second part R0, diluent and FIB adjustment solution to the second part R1, adjust APTT, PT, TT, FIB and AT-Ⅲ values ​​to the target values, and obtain level 2 matrix solution 1, wherein the target values ​​of APTT are 45-150 sec, PT is 15-74 sec, TT is 20-50 sec, FIB is 0.6-2 mg / L, and AT-Ⅲ is 25-80%; The FIB adjustment solution is human plasma concentrate, and its addition amount is 0~50% of the total volume of R0 and R1; the addition amount of PP0 or PP1 accounts for 0~20% of the total volume; the diluent accounts for 0~60% of the total volume; and R0 accounts for 0~20% of the total volume.

[0054] (5) Add DD stock solution and FDP stock solution to Level 1 Matrix Solution 1, and adjust the concentrations of DD and FDP in Level 1 Matrix Solution 1 to the target values ​​of Level 1 Matrix Solution 2, DD is 0.1-0.9 μg / L (FEU), and FDP is 2.2-5.5 μg / L; Then add AT-Ⅲ concentrate to level 1 matrix solution 2, and adjust the AT-Ⅲ value to be consistent with normal human mixed plasma, thus obtaining the seven-item coagulation composite quality control product level 1, in which the AT-Ⅲ target value is 80~120%; Add DD stock solution and FDP stock solution to level 2 matrix solution 1, adjust the concentrations of DD and FDP in level 2 matrix solution 1 to the target values ​​to obtain the coagulation seven-item composite quality control product level 2, where the target values ​​of DD are 0.9~30μg / L and FDP are 5.5~55μg / L; (6) The two-level activated coagulation test reagent quality control product is obtained by subpackaging and freeze-drying.

[0055] The range required for coagulation quality control level 1 is prothrombin time 10~15s, activated partial thromboplastin time 21~45s, fibrinogen concentration 2~4 g / L, antithrombin III activity 80~120%, D-dimer concentration 0.1~0.9 μg / mL (FEU), fibrin degradation product concentration 2.5~5.5μg / mL, and thrombin time 10~20s.

[0056] The range required for coagulation quality control level 2 is prothrombin time 15~74s, activated partial thromboplastin time 45~150s, fibrinogen concentration 0.6~2 g / L, antithrombin III activity 5~80%, D-dimer concentration 0.9~30 μg / mL (FEU), fibrin degradation product concentration 5.5~55 μg / mL, and thrombin time 20~50s.

[0057] The preparation method of the seven-item composite quality control product of coagulation can be adjusted to obtain level 1 quality control products (any index of APTT, PT, TT, FIB, AT-III, DD and FDP can be adjusted to any target value within the normal range) and level 2 quality control products (any index of the seven indexes can be adjusted to any target value within the abnormal value range).

[0058] The two-level activated coagulation seven-item composite quality control product prepared by the method of the present invention.

[0059] The required range of the intermediate indicators of the seven-item composite quality control product level 1 for coagulation is: APTT 25~31s, PT 10.8~13s, TT14~18s, Fib 2.2~3.2g / L, AT-Ⅲ 86~105%, DD 0.3~0.5 ug / ml (FEU), FDP 2.5~5ug / ml.

[0060] The required range of indicators in level 2 of the seven-item composite quality control product for coagulation is: APTT 48~65s, PT 28~36s, TT 28~50s, Fib 1.2~1.7g / L, AT-Ⅲ 30~50%, DD 2.0~3.0ug / ml (FEU), FDP 15~20ug / ml.

[0061] The fixed components and volume contents of coagulation quality control level 1 are: R0 36~81%, R1 0~45%, FIB adjustment solution 0~8.2%, PP0 0~8.2%, FDP 0~0.73%, DD 0~0.73%, AT III 0~0.3%.

[0062] The fixed components and volume contents of the seven-item composite quality control product level 2 for coagulation are: R1 14~37%, diluent 37~59%, FIB adjustment solution 0~3%, R0 0~11%, PP1 0~15%, FDP 0~0.73%, and DD 0~0.73%.

[0063] The present invention has a stable and reliable process; any index of the level 1 quality control product APTT, PT, TT, FIB, AT-III, DD and FDP can be adjusted to any target value within the normal range, and any index of the seven indicators of the level 2 quality control product can be adjusted to any target value within the abnormal value range, which can meet the target value requirements of various coagulation detection systems; the freeze-dried product has good stability after reconstitution; and the quality control product has excellent uniformity. The specific implementation steps are as follows: Example 1

[0064] This embodiment provides a method for preparing a seven-item coagulation composite quality control product (level 1), comprising the following steps: (1) The buffer stabilizing protective agent contains: 480 g of D-trehalose, 50 g of glycine, 120 g of HEPES FREE ACID, add about 500 ml of purified water and heat to dissolve; then add 10 ml of Proclin 300.

[0065] (2) Add 60 ml of aluminum hydroxide colloid to 600 ml of human plasma and 30 ml of aluminum hydroxide colloid to 300 ml of PP, respectively, and stir for 5 minutes, centrifuge at 4000 rpm for 50 minutes to obtain the supernatant. Take 580 ml of the adsorbed human plasma supernatant and add 58 ml of the protective agent to obtain R1, and take 280 ml of the adsorbed PP supernatant and add 28 ml of the protective agent to obtain PP1.

[0066] The plasma is human mixed plasma anticoagulated with sodium citrate and animal plasma PP anticoagulated with sodium citrate. 1 part of sodium citrate and 9 parts of fresh blood in vitro (volume ratio = 1:9) are mixed and centrifuged at 4000rpm for 10 minutes. The clear liquid on the upper layer is plasma.

[0067] (3) Take 600 ml of human plasma and add 60 ml of protective agent to obtain R0; take 100 ml of PP and add 5.56 ml of protective agent to obtain PP0.

[0068] (4) FIB conditioning solution: Weigh 20 g of human plasma concentrate, add 400 ml of purified water, centrifuge at 4000 rpm at 4°C for 20 min, and collect the supernatant; (5) Add 100 ml of adsorbed human plasma, 20 ml of FIB conditioning solution 1, and 15 ml of PP0 to 200 ml of R0 and mix well. Add 16.5 μl of FDP stock solution and 5.2 μl of DD stock solution and mix well. Add 1500 μl of AT-Ⅲ. The ingredients are shown in Table 1 below: Table 1 raw material R0 R1 FIB conditioning fluid PP0 FDP DD AT Ⅲ Dosage 200ml 100ml 20ml 15ml 16.5μl 5.2μl 1500μl Proportion 59.5% 29.8% 5.95% 4.5% 0.005% 0.0015% 0.25% (6) After packaging, the mixture was freeze-dried, reconstituted with water and tested. The results are shown in Table 1.

[0069] The results showed that APTT, PT, TT, FIB, AT-III, FDP and DD were within the range of the seven-item composite quality control product for coagulation (level 1), meeting the requirements for normal quality control products.

[0070] Example 2 This example provides a preparation of a seven-item coagulation composite quality control product (level 2), which is different from Example 1 in that 200 ml of R1 is mixed with 300 ml of diluent, 10 ml of FIB adjustment solution, 30 ml of R0, and 80 ml of PP1, and 50 μl of FDP stock solution and 400 μl of DD stock solution are added and mixed. The content of each component is shown in Table 2 below: Table 2 raw material R1 Dilution FIB conditioning fluid R0 PP1 FDP DD Dosage 200ml 300ml 10ml 30ml 80ml 50μl 400 μl Proportion 32% 48.4% 1.6% 4.8% 12.9% 0.008% 0.064% After freeze-drying, the samples were reconstituted with water and tested. The results are shown in Table 3.

[0071] Table 3 Test results of Examples 1 and 2

[0072] The results showed that APTT, PT, TT, FIB, AT-III, FDP and DD were within the range of the seven-item composite quality control product for coagulation (level 2), meeting the requirements for abnormal quality control products.

[0073] Example 3 This example provides a preparation of a seven-item coagulation composite quality control product (level 2), which is different from Example 1 in that 110 ml of adsorbed human plasma, 25 ml of FIB adjustment solution 1, and 15 ml of PP0 are added to 200 ml of R0 and mixed, 16.5 μl of FDP stock solution and 5.2 μl of DD stock solution are added and mixed, and 850 μl of AT-III is added. The components are shown in Table 4 below: Table 4 R0 R1 FIB conditioning fluid PP0 FDP DD AT-Ⅲ 200ml 110ml 30ml 16ml 16.5μl 5.2μl 850μl 56% 30.8% 7% 4.48% 0.0046% 0.0015% 0.24% Example 4

[0074] This example provides a preparation of a seven-item coagulation composite quality control product (level 2), which is different from Example 1 in that 200 ml of R1 is mixed with 330 ml of diluent, 13 ml of FIB adjustment solution, 34 ml of R0, and 85 ml of PP1, and 50 μl of FDP stock solution and 400 μl of DD stock solution are added and mixed. The components are shown in Table 5 below: Table 5 R1 Dilution FIB conditioning fluid R0 PP1 FDP DD 200ml 330ml 13ml 34ml 85ml 50μl 400 μl 30.2% 49.8% 1.96% 5.13% 12.8% 0.0075% 0.06% After the samples of Examples 3 and 4 were freeze-dried, they were reconstituted with water and tested. The results are shown in Table 6: Table 6 Test results of Examples 3 and 4

[0075] Comparative Examples 1 and 3, after mixing the above different raw material ratios, the APTT, PT, TT and FIB index values ​​can be satisfactorily adjusted within the normal range. Comparative Examples 2 and 4, after mixing the above different raw material ratios, the APTT, PT, TT and FIB index values ​​can be satisfactorily adjusted within the abnormal value range.

[0076] The above diluents can be one or more of D-trehalose, sodium chloride, HEPES FREE ACID, dihydrate. Trisodium citrate. Among them, the percentage concentration range of D-trehalose is 0~100g / L, the percentage concentration range of sodium chloride is 0~50g / L; the percentage concentration range of HEPES FREE ACID is 0~30g / L; the percentage concentration range of dihydrate. Trisodium citrate is 0~5g / L, and the same applies to the following.

[0077] Test 1: Uniformity test of different samples.

[0078] The examples 1 and 2 of the present invention and the commercially available control products (normal) (normal coagulation control product, abnormal coagulation control product (no TT), buffer matrix normal DD control product, buffer matrix abnormal DD control product, buffer matrix normal FDP control product, buffer matrix abnormal FDP control product) were accurately reconstituted with the indicated volume of purified water, and the homogeneity test was performed according to the following method: Take 10 bottles of this product and use the APTT, PT, TT, Fib, AT-Ⅲ, FDP, and D-Dimer detection kits respectively according to their respective instructions and the instructions of the applicable coagulation analyzer. Measure each bottle once (APTT, PT, TT, FIB, AT-Ⅲ, FDP, D-D), and calculate the average value of the test results according to formula 1 (1) and the standard deviation of the test results according to formula 2 (S1); mix the above 10 bottles of reconstituted products and continuously test 10 times (APTT, PT, TT, FIB, AT-Ⅲ, FDP, D-D), and calculate the average value of the test results according to formula 1 (2) and the standard deviation of the test results according to formula 2 (S2); calculate the coefficient of variation within the bottle CV within (APTT, PT, TT, FIB, AT-Ⅲ, FDP, D-D) according to formula 3; calculate the coefficient of variation between bottles CV between (APTT, PT, TT, FIB, AT-Ⅲ, FDP, DD) according to formula 4 and formula 5.

[0079] Formula 1 ; Formula 2 ;

[0080] Formula 3 ;

[0081] Formula 4 ;

[0082] Formula 5 (When S1 < S2, let CV between = 0); In the formula: ----Average value; S----Standard deviation; n----Number of measurements; CV----Coefficient of variation; xi----The i-th measured value of the specified parameter. The formula is from the "General Requirements for Quality Control Substances for In Vitro Diagnostic Reagents" YY / T1652-2019. The results are shown in Table 7 below: Table 7 Detection Results of the Homogeneity of Different Samples

[0083] From the above results, it can be seen that for all items in Examples 1 and 2, CV between and CV within are less than those of the commercially available quality control products, and for CV between of the corresponding items, it proves that the repeatability and precision of the quality control products of the present invention are better than those of the commercially available quality control products.

[0084] Test Two: Detection of the Stability of Different Samples.

[0085] The quality control products of Examples 1 and 2 of the present invention and commercially available quality control products (normal) (normal coagulation quality control product, abnormal coagulation quality control product (no TT), buffer matrix normal DD quality control product, buffer matrix abnormal DD quality control product, buffer matrix normal FDP quality control product, buffer matrix abnormal FDP quality control product) were accurately reconstituted with the indicated volume of purified water and stored under refrigeration (2°C-8°C). Samples were taken every 0h, 4h, 6h, 8h, 17h, 24h, and 48h to detect APTT, PT, TT, FIB, ATIII, DD, and FDP. The stability deviation was calculated with 0h as a reference, and the results are shown in Tables 4 to 7.

[0086] (Stability deviation = ). The results are shown in Tables 8 to 11 below: Table 8 Stability deviation of reconstitution of quality control products in Example 1

[0087] Table 9 Stability deviation of reconstitution of quality control products in Example 2

[0088] Table 10 Reconstitution stability deviation of normal quality control products on the market

[0089] Table 11 Reconstitution stability deviation of abnormal quality control products on the market

[0090] It can be seen from the results in Tables 8 to 11 above that the stability deviation results of all items in Examples 1 and 2 are smaller than those of commercially available quality control products, proving that the stability of the quality control product of the present invention is better than that of commercially available quality control products.

[0091] Experiment 3 Taking Example 1 as an example, set control groups 1-6, the influence of different contents of each component on the results The other contents are the same as those in Example 1, except that the contents of the components are different: 1. A control group 1 was set. The difference from Example 1 was that the amount of FDP stock solution was changed from 16.5 μl to 0 μl, 30 μl, 50 μl, 70 μl, and 100 μl. The test results are shown in Table 12 below.

[0092] Table 12 FDP stock solution dosage and FDP measured value FDP dosage FDP (ug / ml) DD (ug / ml) APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0μl 1.78 0.33 30.93 12.07 12.34 2.89 88.57 30μl 5.21 0.38 30.86 11.95 15.98 2.91 88.46 50μl 9.08 0.36 30.25 12.07 16.15 2.94 88.14 70μl 12.43 0.39 31.25 12.25 16.17 2.92 88.32 100μl 20 0.37 30.97 12.03 16.27 2.97 88.47 The results in Table 12 show that when the dosage of other components remains unchanged, as the dosage of FDP stock solution increases within the range of 0-100 μl, the FDP detection value increases accordingly, and the other six detection values ​​do not change significantly.

[0093] 2. Set up control group 2. The difference from Example 1 is that the dosage of DD stock solution is changed from 5.2 μl to 0 μl, 10 μl, 30 μl, 70 μl, and 100 μl. The test results are shown in Table 13 below.

[0094] Table 13 DD stock solution dosage and DD measured value DD dosage DD (ug / ml) FDP (ug / ml) APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0μl 0.22 3.01 30.86 12.21 16.26 2.98 88.31 10μl 0.43 2.96 30.76 12.17 16.17 2.95 88.37 30μl 0.84 3.00 30.94 12.18 16.21 2.93 89.06 70μl 1.74 2.95 30.84 12.09 16.24 2.91 88.59 100μl 2.45 2.97 30.79 12.19 16.23 2.94 88.34 The results in Table 13 show that when the dosage of other components remains unchanged, as the dosage of DD stock solution increases within the range of 0~100μl, the DD detection value increases accordingly, and the other six detection values ​​do not change significantly.

[0095] 3. Set up control group 3. The difference from Example 1 is that the dosage of AT-III concentrate is changed from 1500 μl to 0 μl, 300 μl, 900 μl, 1500 μl, 2100 μl, and 2700 μl. The test results are shown in Table 14 below.

[0096] Table 14 AT-III concentrate dosage and AT-III measured values AT-Ⅲ dosage AT-Ⅲ(%) DD (ug / ml) FDP (ug / ml) APTT(s) PT(s) TT(s) Fib (g / L) 0 80.01 0.33 3.00 30.81 12.16 16.25 2.94 300μl 82.42 0.32 3.01 30.79 12.13 16.23 2.95 900μl 87.65 0.33 2.94 30.83 12.15 16.13 2.97 1500μl 92.10 0.32 3.03 30.87 12.18 16.26 2.93 2100μl 96.94 0.33 2.99 30.46 12.19 16.24 2.96 2700μl 101.77 0.34 2.99 30.52 12.20 16.27 2.93 From the results in Table 14, it can be seen that when no concentrate is added, the AT-III activity is equivalent to about 80% of that of a normal person. When the dosage of other ingredients remains unchanged, as the dosage of AT-III concentrate increases within the range of 0-2700 μl, the AT-III test value increases accordingly, and the other six test values ​​do not change significantly.

[0097] 4. Set up control group 5. The difference from Example 1 is that AT-III concentrate, FDP stock solution and DD stock solution are not added, and the concentration of FIB regulating solution is changed from 50 g / dL to 0 g / dL, 20 g / dL, 70 g / dL, 100 g / dL and 130 g / dL. The results are shown in Table 15 below.

[0098] Table 15 Effect of FIB regulating solution concentration changes on coagulation activity FIB conditioning solution concentration APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0 g / dL 34.72 15.21 16.69 2.23 80.23 20 g / dL 32.51 14.49 16.45 2.46 80.45 50 g / dL 30.90 12.14 16.20 2.96 80.92 70 g / dL 29.04 11.38 16.11 3.34 81.02 100 g / dL 28.23 11.04 15.93 3.87 81.24 From the results in Table 15, it can be seen that when the dosage of other components remains unchanged, the volume of FIB adjusting solution remains unchanged, the concentration of FIB adjusting solution increases within the range of 0~130g / dL, the FIB detection value increases accordingly, the APTT, PT, and TT measurement values ​​gradually shorten, and the AT-III measurement value does not change significantly.

[0099] 5. Set up control group 5. The difference from Example 1 is that no AT-III concentrate, FDP stock solution, or DD stock solution is added, and the amount of PPO is changed from 15 mL to 0 mL, 7.5 mL, 20 mL, 40 mL, and 60 mL. The results are shown in Table 16 below.

[0100] Table 16 Effect of PP0 dosage changes on coagulation activity PP0 dosage APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0 mL 34.93 15.69 15.53 2.94 81.09 7.5 mL 32.62 14.76 16.26 2.89 81.07 15 mL 30.99 12.38 16.47 2.96 80.92 20 mL 29.35 11.57 16.63 2.91 81.12 40 mL 28.59 11.24 16.73 2.88 80.84 60 mL 26.72 10.81 16.95 2.93 81.16 From the results in Table 16, it can be seen that when the dosage of other components remains unchanged, as the dosage of PP0 increases within the range of 0~40mL, the APTT and PT test values ​​are shortened, the TT test value is slightly prolonged, and the FIB and AT-III test values ​​have no obvious changes.

[0101] 6. A control group 6 was set up. The difference from Example 1 was that no AT-III concentrate, FDP stock solution, or DD stock solution was added, and the total amount of R0 and R1 remained unchanged. The change in the ratio of the two and the test results were as shown in Table 17: Table 17 Effect of changes in the volume ratio of R0 to R1 on coagulation activity R0 dosage R1 dosage APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 300 mL 0 mL 26.27 10.80 16.44 2.92 80.85 250 mL 50 mL 28.18 11.27 16.51 2.91 80.73 200 mL 100 mL 30.99 12.38 16.47 2.96 80.92 150 mL 150 mL 32.57 15.31 16.38 2.85 81.04 50 mL 200 mL 46.39 24.16 16.42 2.93 80.79 0 mL 300 mL 57.81 26.94 16.53 2.89 81.10 The results in Table 17 show that when the dosage of other components remains unchanged and the total volume of R0 and R1 remains unchanged, as the dosage of R0 decreases and the dosage of R1 gradually increases, the APTT and PT test values ​​are extended, while the TT, FIB, and AT-III test values ​​do not change significantly.

[0102] Experiment 4 Taking Example 2 as an example, set control groups 7-13, the influence of different contents of each component on the results The other contents are the same as those in Example 2, except that the contents of the components are different: 1. A control group 7 was set up. The difference from Example 2 was that the amount of FDP stock solution used was changed from 50 μl to 0 μl, 25 μl, 50 μl, 75 μl, and 100 μl. The test results are shown in Table 18 below.

[0103] Table 18 FDP stock solution dosage and FDP measured value FDP dosage FDP (ug / ml) DD (ug / ml) APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0μl 1.04 2.61 57.33 39.70 31.35 1.42 38.71 25μl 10.25 2.59 57.45 39.76 31.42 1.41 38.74 50μl 18.06 2.62 57.48 39.71 31.49 1.44 38.69 75μl 26.32 2.58 57.43 39.61 31.36 1.43 38.62 100μl 35.52 2.63 57.49 37.82 31.47 1.43 38.65 The results in Table 18 show that when the dosage of other components remains unchanged, as the dosage of FDP stock solution increases within the range of 0-100 μl, the FDP detection value increases accordingly, and the other six detection values ​​do not change significantly.

[0104] 2. Set up control group 8. The difference from Example 2 is that the dosage of DD stock solution is changed from 400 μl to 0 μl, 200 μl, 700 μl, and 1000 μl. The test results are shown in Table 19 below.

[0105] Table 19, DD stock solution dosage and DD measured value DD dosage DD (ug / ml) FDP (ug / ml) APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0μl 0.41 18.09 57.47 39.68 31.53 1.43 38.56 200 μl 1.28 18.05 57.39 39.76 31.42 1.44 38.72 400μl 2.62 18.06 57.48 39.71 31.49 1.44 38.69 700μl 4.94 18.10 57.51 39.68 31.45 1.45 38.64 1000μl 6.48 18.07 57.46 37.78 31.49 1.46 38.62 The results in Table 19 show that when the dosage of other components remains unchanged, as the dosage of DD stock solution increases within the range of 0~1000μl, the DD detection value increases accordingly, and the other six detection values ​​do not change significantly.

[0106] 3. A control group 9 was set up. The difference from Example 2 was that no FDP stock solution or DD stock solution was added, and the amount of FIB regulating solution was changed from 10 mL to 0 mL, 5 mL, 20 mL, 30 mL, and 40 mL. The results are shown in Table 20 below.

[0107] Table 20 Effect of changes in the amount of FIB regulating solution on coagulation activity FIB conditioning solution volume Fib (g / L) APTT(s) PT(s) TT(s) AT-Ⅲ(%) 0mL 0.61 89.64 61.28 49.26 38.26 5mL 1.03 68.43 50.62 39.47 38.47 10mL 1.44 57.48 39.71 31.49 38.69 20mL 2.25 46.82 32.59 24.51 38.79 30mL 3.09 41.19 27.28 19.43 38.94 40mL 3.95 39.49 21.57 17.31 39.12 From the results in Table 20, it can be seen that when the dosage of other components remains unchanged, the dosage of FIB adjustment solution increases within the range of 0~40mL, the FIB detection value increases accordingly, the APTT, PT, and TT measurement values ​​gradually shorten, and the AT-III measurement value does not change significantly.

[0108] 4. Set up control group 10. The difference from Example 2 is that no FDP stock solution and DD stock solution are added, and the amount of PP1 is changed from 80 mL to 0 mL, 40 mL, 120 mL, and 160 mL. The results are shown in Table 21 below.

[0109] Table 21 Effect of PP1 dosage changes on coagulation activity PP1 dosage APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0 mL 110.25 100.24 29.43 1.42 38.58 40mL 81.72 65.13 30.47 1.45 38.74 80 mL 57.48 39.71 31.49 1.44 38.69 120 mL 41.46 30.45 32.46 1.43 38.72 160 mL 29.10 25.24 34.73 1.46 38.63 From the results in Table 21, it can be seen that when the dosage of other components remains unchanged, as the dosage of PP1 increases within the range of 0~160mL, the APTT and PT test values ​​are shortened, the TT test value is slightly prolonged, and the FIB and AT-III test values ​​have no obvious changes.

[0110] 5. Set up control group 11. The difference from Example 2 is that no FDP stock solution and DD stock solution are added, and the amount of R0 is changed from 30mL to 0mL, 15mL, 50mL, and 70mL. The results are shown in Table 22 below.

[0111] Table 22 Effect of R0 dosage changes on coagulation activity R0 dosage APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 0 mL 71.23 50.27 31.42 1.38 35.51 15mL 60.24 41.13 31.47 1.41 37.02 30 mL 57.48 39.71 31.49 1.44 38.69 50mL 52.43 37.16 31.51 1.46 39.51 70mL 48.27 35.28 31.43 1.49 40.26 The results in Table 22 show that when the dosage of other components remains unchanged, as the dosage of R0 increases in the range of 0~70mL, the APTT and PT test values ​​decrease accordingly, while the TT, FIB, and AT-III test values ​​do not change significantly.

[0112] 6. A control group 12 was set up. The difference from Example 2 was that no FDP stock solution and DD stock solution were added, and the total amount of R1 and diluent remained unchanged. The change in the ratio of the two and the test results were shown in Table 23 below: Table 23 Effect of the volume ratio of R1 to diluent on coagulation activity R1 dosage Dilution amount APTT(s) PT(s) TT(s) Fib (g / L) AT-Ⅲ(%) 50 mL 450 mL 155.43 90.19 65.27 0.84 14.47 100 mL 400 mL 80.24 67.94 45.68 1.16 24.34 200 mL 300 mL 57.48 39.71 31.49 1.44 38.69 300 mL 200 mL 49.81 35.47 30.18 1.68 52.41 400mL 100 mL 42.37 33.26 29.42 1.79 66.39 500 mL 0 mL 39.81 31.20 28.37 1.84 82.56 The results in Table 23 show that when the dosage of other components remains unchanged and the total volume of R1 and diluent remains unchanged, as the dosage of R1 increases and the dosage of R1 gradually decreases, the APTT, PT, and TT test values ​​shorten accordingly, and the FIB and AT-III test values ​​gradually increase.

[0113] The above implementation / test examples are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A method for preparing a seven-item composite quality control product for coagulation, characterized in that: The following steps are involved: (1) Obtaining sodium citrate anticoagulated human mixed plasma and sodium citrate anticoagulated animal plasma PP; (2) Part of the human mixed plasma and PP were adsorbed by aluminum hydroxide colloid, and the supernatant was collected by centrifugation to obtain adsorbed human mixed plasma and adsorbed PP; (3) Add buffering and stabilizing protective agents to another part of human mixed plasma and PP, as well as adsorbed human mixed plasma and adsorbed PP, to obtain the corresponding R0, PP0, R1 and PP1; (4) Divide R0 and R1 into two parts: add PP0, the first part R1 and FIB adjustment solution to the first part R0, adjust the FIB, APTT, PT and TT values ​​to be consistent with normal human mixed plasma, and obtain level 1 matrix solution 1, in which the target values ​​of APTT are 21~45sec, PT are 10~15sec, TT are 10~20sec, and FIB are 2~4mg / L; Add PP1, second part R0, diluent and FIB adjustment solution to the second part R1, adjust APTT, PT, TT, FIB and AT-Ⅲ values ​​to the target values, and obtain level 2 matrix solution 1, wherein the target values ​​of APTT are 45-150 sec, PT is 15-74 sec, TT is 20-50 sec, FIB is 0.6-2 mg / L, and AT-Ⅲ is 25-80%; (5) Add DD stock solution and FDP stock solution to Level 1 matrix solution 1, and adjust the concentrations of DD and FDP in Level 1 matrix solution 1 to the target values ​​of Level 1 matrix solution 2, DD is 0.1~0.9μg / L, and FDP is 2.2~5.5μg / L; then add AT-Ⅲ concentrate to Level 1 matrix solution 2, and adjust the AT-Ⅲ value to be consistent with normal human mixed plasma, so as to obtain the seven-item composite quality control product Level 1 of coagulation, in which the target value of AT-Ⅲ is 80~120%; Add DD stock solution and FDP stock solution to level 2 matrix solution 1, adjust the concentrations of DD and FDP in level 2 matrix solution 1 to the target values ​​to obtain the coagulation seven-item composite quality control product level 2, where the target values ​​of DD are 0.9~30μg / L and FDP are 5.5~55μg / L; (6) Subpackaging and freeze-drying to obtain two levels of activated coagulation test reagent quality control products.

2. The method for preparing a seven-item composite quality control product for coagulation according to claim 1, characterized in that: The amount of aluminum hydroxide colloid used in step (2) (V / V) is 1% to 10%.

3. The method for preparing a seven-item composite quality control product for coagulation according to claim 1, characterized in that: In the step (3), the buffer stabilizing protective agent is one or more of trehalose, glycine, HEPES, sodium chloride, trisodium citrate, Tris, and Proclin300.

4. The method for preparing a seven-item composite quality control product for coagulation according to claim 3, characterized in that: The trehalose concentration range is 50-500 g / L, the glycine concentration range is 0-160 g / L, the HEPES concentration range is 100-250 g / L, the sodium chloride concentration range is 0-10 g / L, the trisodium citrate concentration range is 0-5 g / L, the Tris concentration range is 0-5 g / L, and the Proclin300 concentration range is 1 / 1000 to 10; the added amount of the buffer stabilizing protective agent is 4% to 10%.

5. The method for preparing a seven-item composite quality control product for coagulation according to claim 1, characterized in that: In step (4), the FIB regulating solution is human plasma concentrate, and its addition amount is 0-50% of the total volume of R0 and R1; the addition amount of PP0 or PP1 accounts for 0-20% of the total volume; the diluent accounts for 0-60% of the total volume; and R0 accounts for 0-20% of the total volume.

6. The method for preparing a seven-item composite quality control product for coagulation according to claim 1, characterized in that: The seven-item composite quality control product level 1 of coagulation in step (5) has a prothrombin time of 10-15 s, an activated partial thromboplastin time of 21-45 s, a fibrinogen concentration of 2-4 g / L, an antithrombin III activity of 80-120%, a D-dimer concentration of 0.1-0.9 μg / mL, a fibrin degradation product concentration of 2.5-5.5 μg / mL, and a thrombin time of 10-20 s.

7. The method for preparing a seven-item composite quality control product for coagulation according to claim 1, characterized in that: The seven-item composite quality control product level 2 of the coagulation test in step (5) has a prothrombin time of 15-74 s, an activated partial thromboplastin time of 45-150 s, a fibrinogen concentration of 0.6-2 g / L, an antithrombin III activity of 5-80%, a D-dimer concentration of 0.9-3.0 μg / mL, a fibrin degradation product concentration of 5.5-55 μg / mL, and a thrombin time of 20-50 s.

8. A seven-item composite quality control product for coagulation prepared by the method according to any one of claims 1 to 7.

9. A seven-item composite quality control product for coagulation according to claim 8, characterized in that: The fixed components and volume contents of the seven-item composite quality control product level 1 of coagulation are: R0 36-81%, R1 0-45%, FIB adjustment solution 0-8.2%, PP0 0-8.2%, FDP 0-0.73%, DD 0-0.73%, AT III 0-0.3%; The seven-item composite quality control level 1 indicators of coagulation are: APTT 25~31s, PT 10.8~13s, TT 14~18s, FIB2.2~3.2g / L, AT-Ⅲ 86~105%, DD 0.3~0.5 ug / ml, and FDP 2.5~5ug / ml.

10. A seven-item composite quality control product for coagulation according to claim 8, characterized in that: The fixed components and volume contents of the seven-item composite quality control product level 2 for coagulation are: R1 14-37%, diluent 37-59%, FIB adjustment solution 0-3%, R0 0-11%, PP1 0-15%, FDP 0-0.73%, DD 0-0.73%; The seven-item composite quality control product level 2 indicators of coagulation are: APTT 48~65s, PT 28~36s, TT 28~50s, FIB 1.2~1.7g / L, AT-Ⅲ 30~50%, DD 2.0~3.0ug / ml, and FDP 15~20ug / ml.

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