A coagulation seven-item composite quality control product and its preparation method

Abstract

The present invention belongs to the technical field of coagulation detection, and particularly relates to a seven - item coagulation composite quality control product and a preparation method thereof. The quality control product has two levels, namely level 1 and level 2. The seven indicators include APTT, PT, TT, FIB, AT - III, D - Dimer and FDP. In the present invention, the process is stable and reliable; for the level 1 quality control product, any one of the indicators of APTT, PT, TT, FIB, AT - III, D - D and FDP can be adjusted to any target value within the normal range, and for the level 2 quality control product, any one of the seven indicators 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 stability of the freeze - dried product after reconstitution is good; and the uniformity of the quality control product is excellent.

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 plasma to coagulate after adding standardized thrombin. Since fibrin(ogen) degradation products (FDP) can prolong TT, it can be used as a screening test for the fibrinolytic system, reflecting the conversion of fibrinogen to fibrin in the common coagulation pathway. Clinically, phenomena that can lead to prolonged TT include: increased heparin or the presence of heparin-like anticoagulant substances, increased fibrinogen degradation products, disseminated intravascular coagulation (DIC), hypo(afibrinogenemia), etc.

[0007] FIB is the precursor of fibrin, an important protein fiber involved in the processes of blood coagulation and hemostasis. Changes in its concentration have an important impact on blood coagulation function. Clinically, increased FIB is an important risk factor for various thrombotic diseases, 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 commonly seen in DIC, primary fibrinolysis, severe hepatitis and cirrhosis, hypofibrinogenemia; it can also be seen during the use of fibrinolytic drugs (antithrombotic enzyme, defibrase) and thrombolytic therapy.

[0008] AT-III is an important anticoagulant substance in the body, which can inhibit the activities of thrombin and coagulation factors IXa, Xa, XIa, XIIa, etc., thereby regulating the coagulation process and maintaining the balance between coagulation and anticoagulation. Clinically, deficiency or reduced activity of AT-III can lead to a hypercoagulable state of the blood, prone to thrombosis, commonly seen in congenital AT-III deficiency, DIC, liver diseases, nephrotic syndrome and other disease processes. In addition, in the monitoring of anticoagulant therapy, the effects of certain anticoagulant drugs (such as heparin) rely on AT-III. If the AT-III level is too low, the anticoagulant effect of heparin may not be fully exerted, and at this time, it is necessary to supplement AT-III or adjust the anticoagulant therapy plan.

[0009] FDP is the degradation product produced after fibrinogen and fibrin are decomposed. The level of 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 in the body and the activation of the fibrinolytic system.

[0011] Therefore, in the diagnosis, detection and treatment of hematological diseases, especially in recent years when thrombus-related diseases have received increasing attention, the seven - item coagulation test will play an important role in the monitoring, prevention and treatment of diseases such as thrombosis.

[0012] To improve clinical monitoring efficiency and reduce monitoring costs, more and more manufacturers of coagulation testing equipment and reagents are focusing on the development of an integrated detection system for the above seven coagulation tests. As a result, there is also a growing demand for quality control products for the seven coagulation tests. In the past, coagulation quality control products mainly targeted the detection of single or the first 4-5 coagulation indicators, and the development of composite quality control products for the seven coagulation tests was relatively rare.

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

[0014] The specification of Chinese Patent CN118425538A records an abnormal value seven coagulation composite quality control product and its preparation method. Using rabbit plasma as the matrix plasma, mainly using hirudin as an anticoagulant to regulate relevant coagulation time items, an abnormal value quality control product is prepared, which can be used for PT, APTT, TT, FIB, AT-Ⅲ, D-Dimer, and FDP items at the same time. In this patent, rabbit plasma is completely used as the matrix plasma, which may be different from the human blood matrix environment during actual detection. When conditions permit, it is most ideal to use human-derived plasma as the main component as much as possible. The addition of hirudin, a non-human-derived material, 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 included, so this method has great limitations.

[0015] The coagulation process is a process in which multiple factors (coagulation factors, other conditions) act together and restrict each other, and finally is reflected in items such as PT, APTT, TT, FIB, AT-Ⅲ, D-Dimer, and FDP. It is difficult to debug a single item alone without changing other items at all. In addition, due to the existence of differences in biological raw materials, there may be large batch-to-batch differences in quality control products, and even the phenomenon that the target value cannot be successfully debugged may occur.

[0016] The specification of Chinese Patent CN117741166A records that serum is used as one of the main components of the mixed matrix. Since a series of coagulation factors are activated during the formation of serum, these activated coagulation factors will bring uncertainty to the long-term stability of multiple coagulation quality control products. Therefore, human serum should be avoided or mixed as much as possible in the matrix of multiple quality control products. And the preparation method of this patent by adding coagulation factors increases the cost required for the purification of coagulation factors.

[0017] Therefore, there is an urgent need for a preparation method of a seven - item coagulation composite quality control product, which can adapt to any human plasma raw material, and the quality control product prepared by this method is stable for a long time, and the content or measured value of each item can be adjusted arbitrarily within the normal range or abnormal value range to meet the detection requirements of multiple systems or the requirements of quality control. Summary of the Invention

[0018] To solve the above - mentioned technical problems, the present invention provides a seven - item coagulation composite quality control product and its preparation method. Using human plasma as the matrix plasma, it avoids matrix effects, and the preparation process is reliable and highly operable; the prepared quality control product can meet a wide range of target value requirements and accurately target any target value required for quality control, and at the same time has wide applicability to reagents and instruments and long - term stability.

[0019] A preparation method of a seven - item coagulation composite quality control product in the present invention for solving the above - mentioned technical problems includes the following steps:

[0020] (1) Obtain citrate - anticoagulated human mixed plasma and citrate - anticoagulated animal plasma PP, and set aside;

[0021] Some coagulation factors in plasma are easily inactivated, so the fresher the raw plasma, the better.

[0022] The human mixed plasma used is the plasma of normal people, but in practice, due to factors such as freezing and storage, the coagulation activity decays, and the APTT and PT measured values are prolonged. For PP, the coagulation factor activity is stronger than that of humans, and it has shorter APTT and PT values than humans. The role of animal plasma (PP) is to regulate the APTT and PT of the quality control product; for plasma with partially lost activity, more PP is needed to adjust it to the corresponding APTT and PT values of fresh human mixed plasma.

[0023] For PP, in practical applications, it can be screened through detection, and the APTT measured value is within the range of 14 - 18 seconds.

[0024] (2) Part of the human mixed plasma and PP are respectively adsorbed by aluminum hydroxide (Al(OH)3) colloid, centrifuged to obtain the supernatant, and adsorbed human mixed plasma and adsorbed PP are obtained;

[0025] 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, the measured values of different systems are often inconsistent; the manufacturers of detection systems have their own target values for each index of the quality control product. The method of the present invention is to meet any detection system and adjust the measured values of the seven indexes of the quality control product to the target values.

[0026] (3) Another group of people mixed plasma and PP, and added buffer stabilizing protectants to the adsorbed human mixed plasma and adsorbed PP respectively to obtain corresponding R0, PP0, R1 and PP1.

[0027] (4) Divide R0 and R1 into two parts: Add PP0, the first part of R1 and FIB regulating solution to the first part of R0, and adjust the values of FIB, APTT, PT and TT to be consistent with those of fresh normal human mixed plasma to obtain matrix solution 1 at level 1, where the target values are APTT of 21 - 45 sec, PT of 10 - 15 sec, TT of 10 - 20 sec, and FIB of 2 - 4 mg / L;

[0028] Add PP1, the second part of R0, diluent and FIB regulating solution to the second part of R1, and adjust the values of APTT, PT, TT, FIB, and AT-Ⅲ to the target values to obtain matrix solution 1 at level 2, where the target values are APTT of 45 - 150 sec, PT of 15 - 74 sec, TT of 20 - 50 sec, FIB of 0.6 - 2 mg / L, and AT-Ⅲ of 25 - 80%.

[0029] (5) Add DD stock solution and FDP stock solution to matrix solution 1 at level 1, and adjust the concentrations of DD and FDP in matrix solution 1 at level 1 to the target values to obtain matrix solution 2 at level 1, where DD is 0.1 - 0.9 μg / L (FEU) and FDP is 2.2 - 5.5 μg / L;

[0030] Then add AT-Ⅲ concentrate to matrix solution 2 at level 1, and adjust the value of AT-Ⅲ to be consistent with that of normal human mixed plasma to obtain the coagulation seven-item composite quality control product at level 1, where the target value of AT-Ⅲ is 80 - 120%;

[0031] Add DD stock solution and FDP stock solution to matrix solution 1 at level 2, and adjust the concentrations of DD and FDP in matrix solution 1 at level 2 to the target values to obtain the coagulation seven-item composite quality control product at level 2, where the target values are DD of 0.9 - 30 μg / L and FDP of 5.5 - 55 μg / L.

[0032] (6) Subpackage and freeze-dry to obtain two-level activated coagulation test reagent quality control products.

[0033] Level 1 is the normal value quality control product; level 2 is the abnormal value quality control product.

[0034] In the present invention, there is no need to adjust the pH because the buffer stabilizing protectant has already completed the function of adjusting the pH.

[0035] In the optimized scheme, the dosage (V / V) of aluminum hydroxide colloid in step (2) is 1% - 10%.

[0036] In step (1), the PP is the plasma of one kind of animal among cattle, pigs, sheep and horses; the centrifugation parameters are 4000 rpm at 4 °C for 10 - 50 min.

[0037] Further, in step (3), the buffer stabilizing and protecting agent is one or several of trehalose, glycine, HEPES, sodium chloride, trisodium citrate, Tris and Proclin300.

[0038] Furthermore, the percentage concentration range of glycine is 0 - 160 g / L, the percentage concentration range of HEPES is 100 - 250 g / L, the percentage concentration range of trehalose is 100 - 500 g / L, the percentage concentration range of sodium chloride is 0 - 10 g / L, the percentage concentration range of trisodium citrate is 0 - 5 g / L, the percentage concentration range of Tris is 0 - 5 g / L, and the concentration range of Proclin300 (V / V) is 1‰ to 10‰.

[0039] The addition amount (V / V) of the buffer stabilizing and protecting agent in step (3) is 3% - 7%. By volume ratio, that is, 3 - 7 ml is added to 100 ml.

[0040] 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; R0 accounts for 0 - 20% of the total volume.

[0041] In the present invention, the diluent is one or several of D - trehalose, sodium chloride, HEPES FREE ACID, trisodium citrate dihydrate.

[0042] 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; the percentage concentration range of trisodium citrate dihydrate is 0 - 5 g / L.

[0043] Further, the required range of the coagulation factor VII composite quality control product level 1 is that the prothrombin time is 10 - 15 s, the activated partial thromboplastin time is 21 - 45 s, the fibrinogen concentration is 2 - 4 g / L, the antithrombin III activity is 80 - 120%, the D - dimer concentration is 0.1 - 0.9 μg / mL (FEU), the fibrin degradation product concentration is 2.5 - 5.5 μg / mL, and the thrombin time is 10 - 20 s.

[0044] The range required for the coagulation seven - item composite quality control product at level 2 is as follows: prothrombin time is 15 - 74 s, activated partial thromboplastin time is 45 - 150 s, fibrinogen concentration is 0.6 - 2 g / L, antithrombin III activity is 5 - 80%, D - dimer concentration is 0.9 - 30 μg / mL (FEU), fibrin degradation product concentration is 5.5 - 55 μg / mL, and thrombin time is 20 - 50 s.

[0045] The preparation method of the described coagulation seven - item composite quality control product can be adjusted to obtain level 1 quality control product (any index of APTT, PT, TT, Fib, AT - III, D - D, and FDP can be adjusted to any target value within the normal range), and level 2 quality control product (any index of the seven items can be adjusted to any target value within the abnormal value range).

[0046] In the present invention, the coagulation seven - item composite quality control product prepared by the method described in any one of the above is the two - level activated coagulation seven - item composite quality control product.

[0047] Furthermore, the range of the index requirements for the coagulation seven - item composite quality control product at level 1 is as follows: APTT is 25 - 31 s, PT is 10.8 - 13 s, TT is 14 - 18 s, Fib is 2.2 - 3.2 g / L, AT - Ⅲ is 86 - 105%, D - D is 0.3 - 0.5 μg / ml (FEU), and FDP is 2.5 - 5 μg / ml.

[0048] The range of the index requirements for the coagulation seven - item composite quality control product at level 2 is as follows: APTT is 48 - 65 s, PT is 28 - 36 s, TT is 28 - 50 s, Fib is 1.2 - 1.7 g / L, AT - Ⅲ is 30 - 50%, D - D is 2.0 - 3.0 μg / ml (FEU), and FDP is 15 - 20 μg / ml.

[0049] Furthermore, the fixed components and volume contents of the coagulation seven - item composite quality control product at level 1 are as follows: R0 is 36 - 81%, R1 is 0 - 45%, FIB regulating solution is 0 - 8.2%, PP0 is 0 - 8.2%, FDP is 0 - 0.73%, DD is 0 - 0.73%, and AT Ⅲ is 0 - 0.3%.

[0050] The fixed components and volume contents of the coagulation seven - item composite quality control product at level 2 are as follows: R1 is 14 - 37%, diluent is 37 - 59%, FIB regulating solution is 0 - 3%, R0 is 0 - 11%, PP1 is 0 - 15%, FDP is 0 - 0.73%, and DD is 0 - 0.73%.

[0051] In the present invention, the content of each component has a great influence on the result, and each step is used to facilitate the adjustment of the target value of each item index.

[0052] The coagulation seven - item composite quality control product of the present invention and its preparation method are based on years of experience in researching and producing coagulation series quality control products, and have undergone theoretical and process reliability evaluations, pilot - scale verifications, and mass - production tests.

[0053] The quality control product prepared by the method of the present invention has been tested by various coagulation systems and shows excellent performance in terms of product homogeneity, reconstitution stability, etc.; for the detection of FIB, FDP, and DD that require quantitative detection values, good precision and target - value accuracy are maintained within the detection range; good precision and homogeneity are shown in the detection of levels such as APTT, PT, TT, and AT III. Brief Description of the Drawings

[0054] Figure 1 It is a schematic diagram of the preparation process of the coagulation seven - item composite quality control product at level 1 in the present invention;

[0055] Figure 2 It is a schematic diagram of the preparation process of the coagulation seven - item composite quality control product at level 2 in the present invention. Detailed Embodiments

[0056] The following further elaborates on the present invention in combination with specific embodiments:

[0057] The present invention specifically relates to a preparation method of two - level coagulation seven - item composite quality control products, coagulation seven - item composite quality control product LI, and coagulation seven - item composite quality control product LII. The seven coagulation items are activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), FIB (fibrinogen), antithrombin III (AT - III), fibrinolytic products related to thrombolysis (FDP), and D - dimer (D - Dimer).

[0058] The coagulation seven - item composite quality control product in the present invention is divided into level 1 and level 2 quality control products. The seven indicators include APTT, PT, TT, FIB, AT - III, D - Dimer, and FDP. The preparation method of this coagulation seven - item composite quality control product includes the following steps:

[0059] (1) Obtain citrate - anticoagulated human mixed plasma and citrate - anticoagulated animal plasma for standby;

[0060] (2) Part of the human mixed plasma and PP are respectively adsorbed by aluminum hydroxide (Al(OH)3) colloid, centrifuged to obtain the supernatant, and adsorbed human mixed plasma and adsorbed PP are obtained; the dosage of aluminum hydroxide colloid (V / V) is 1% - 10%; PP is one kind of animal plasma among bovine, porcine, ovine, and equine; the centrifugation parameters are 4°C, 4000 rpm, and centrifugation for 10 - 50 min.

[0061] (3) Another group of people mixed the pooled plasma and PP, and added buffer stabilizing and protecting agents to the adsorbed pooled plasma and adsorbed PP respectively to obtain the corresponding R0, PP0, R1, and PP1;

[0062] The buffer stabilizing and protecting agent is one or more of trehalose, glycine, HEPES, sodium chloride, trisodium citrate, Tris, and Proclin300. The addition amount (V / V) of the buffer stabilizing and protecting agent is 3% - 7%. By volume ratio, that is, 3 - 7 ml is added to 100 ml.

[0063] The percentage concentration range of glycine is 0 - 160 g / L, the percentage concentration range of HEPES is 100 - 250 g / L, the percentage concentration range of trehalose is 100 - 500 g / L, the percentage concentration range of sodium chloride is 0 - 10 g / L, the percentage concentration range of trisodium citrate is 0 - 5 g / L, the percentage concentration range of Tris is 0 - 5 g / L, and the concentration range of Proclin300 (V / V) is 1‰ to 10‰.

[0064] (4) Divide R0 and R1 into two parts: Add PP0, the first part of R1, and FIB adjusting solution to the first part of R0 to adjust the values of FIB, APTT, PT, and TT to be consistent with those of fresh normal pooled plasma, and obtain the level 1 matrix solution 1, where the target values are APTT of 21 - 45 sec, PT of 10 - 15 sec, TT of 10 - 20 sec, and FIB of 2 - 4 mg / L;

[0065] Add PP1, the second part of R0, diluent, and FIB adjusting solution to the second part of R1 to adjust the values of APTT, PT, TT, FIB, and AT - Ⅲ to the target values, and obtain the level 2 matrix solution 1, where the target values are APTT of 45 - 150 sec, PT of 15 - 74 sec, TT of 20 - 50 sec, FIB of 0.6 - 2 mg / L, and AT - Ⅲ of 25 - 80%;

[0066] The FIB adjusting 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; R0 accounts for 0 - 20% of the total volume.

[0067] (5) Add DD stock solution and FDP stock solution to the level 1 matrix solution 1 to adjust the concentrations of DD and FDP in the level 1 matrix solution 1 to the target values to obtain the level 1 matrix solution 2, where DD is 0.1 - 0.9 μg / L (FEU) and FDP is 2.2 - 5.5 μg / L;

[0068] Add AT-Ⅲ concentrate to the level 1 matrix solution 2, and adjust the AT-Ⅲ value to be consistent with that of normal pooled plasma to obtain the level 1 of the seven coagulation factor composite quality control product, where the target value of AT-Ⅲ is 80-120%;

[0069] Add DD stock solution and FDP stock solution to the level 2 matrix solution 1, and adjust the concentrations of DD and FDP in the level 2 matrix solution 1 to the target values to obtain the level 2 of the seven coagulation factor composite quality control product, where the target value of DD is 0.9-30 μg / L and that of FDP is 5.5-55 μg / L;

[0070] (6) Subpackage and lyophilize to obtain the two-level activated coagulation test reagent quality control product.

[0071] The required ranges for the level 1 of the coagulation quality control product are as follows: prothrombin time 10-15 s, activated partial thromboplastin time 21-45 s, 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 2.5-5.5 μg / mL, thrombin time 10-20 s.

[0072] The required ranges for the level 2 of the coagulation quality control product are as follows: prothrombin time 15-74 s, activated partial thromboplastin time 45-150 s, 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, thrombin time 20-50 s.

[0073] The preparation method of the seven coagulation factor composite quality control product can be adjusted to obtain the level 1 quality control product (any index of APTT, PT, TT, Fib, AT-III, D-D and FDP can be adjusted to any target value within the normal range) and the level 2 quality control product (any index of the seven indexes can be adjusted to any target value within the abnormal value range).

[0074] The two-level activated seven coagulation factor composite quality control product prepared by the method in the present invention.

[0075] The required ranges of the indexes in the level 1 of the seven coagulation factor composite quality control product are as follows: APTT 25-31 s, PT 10.8-13 s, TT 14-18 s, Fib 2.2-3.2 g / L, AT-Ⅲ 86-105%, D-D 0.3-0.5 ug / ml (FEU), FDP 2.5-5 ug / ml.

[0076] Range of index requirements in Coagulation Seven - item Composite Quality Control Product Level 2: APTT 48 - 65s, PT 28 - 36s, TT 28 - 50s, Fib 1.2 - 1.7g / L, AT - Ⅲ 30 - 50%, D - D 2.0 - 3.0ug / ml (FEU), FDP 15 - 20ug / ml.

[0077] The fixed components and volume contents of Coagulation Quality Control Product Level 1 are: R0 36 - 81%, R1 0 - 45%, FIB Adjusting Solution 0 - 8.2%, PP0 0 - 8.2%, FDP 0 - 0.73%, DD 0 - 0.73%, AT Ⅲ 0 - 0.3%.

[0078] The fixed components and volume contents of Coagulation Seven - item Composite Quality Control Product Level 2 are: R1 14 - 37%, Diluent 37 - 59%, FIB Adjusting Solution 0 - 3%, R0 0 - 11%, PP1 0 - 15%, FDP 0 - 0.73%, DD 0 - 0.73%.

[0079] In the present invention, the process is stable and reliable; for Level 1 quality control product, any index of APTT, PT, TT, Fib, AT - III, D - D and FDP can be adjusted to any target value within the normal range, and for Level 2 quality control product, any of the seven indexes 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 stability of the freeze - dried product after reconstitution is good; the homogeneity of the quality control product is excellent. The specific implementation steps are as follows:

[0080] Example 1

[0081] This example provides a preparation method of a coagulation seven - item composite quality control product (Level 1), including the following steps:

[0082] (1) The buffer stabilizing and protecting agent contains: 480 grams of D - trehalose, 50 grams of glycine; 120 grams of HEPES FREE ACID; after adding about 500 ml of purified water and heating to dissolve, add 10 ml of Proclin300.

[0083] (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, stir and adsorb for 5 minutes respectively, centrifuge at 4000 rpm for 50 minutes and take the supernatant. Take 580 ml of the adsorbed human plasma supernatant and add 58 ml of the protecting agent to obtain R1, and take 280 ml of the adsorbed PP supernatant and add 28 ml of the protecting agent to obtain PP1.

[0084] The plasma is a mixture of human plasma anticoagulated with sodium citrate and animal plasma PP anticoagulated with sodium citrate. Mix 1 part of sodium citrate and 9 parts of fresh blood in vitro (volume ratio = 1:9), and centrifuge at 4000 rpm for 10 minutes. The clear liquid on the upper layer is the plasma.

[0085] (3) Add 600 ml of human plasma to 60 ml of the protective agent to obtain R0; add 100 ml of PP to 5.56 ml of the protective agent to obtain PP0.

[0086] (4) For the FIB adjustment solution, weigh 20 g of human plasma concentrate, add 400 ml of purified water, centrifuge at 4000 rpm at 4°C for 20 minutes, and collect the supernatant.

[0087] (5) In 200 ml of R0, add 100 ml of adsorbed human plasma, 20 ml of FIB adjustment solution 1, 15 ml of PP0, mix well, add 16.5 μl of FDP stock solution, 5.2 μl of DD stock solution, and mix well. Then add 1500 μl of AT-III. The components are shown in Table 1 below:

[0088] Table 1

[0089] Raw material R0 R1 FIB Adjusting Solution PP0 FDP DD AT Ⅲ Dosage 200ml 100ml 20ml 15ml 16.5 μl 5.2 μl 1500 μl Proportion 59.5% 29.8% 5.95% 4.5% 0.005% 0.0015% 0.25%

[0090] (6) After dispensing, freeze-dry, reconstitute with water and then detect. The results are shown in Table 1.

[0091] The results show that APTT, PT, TT, FIB, AT-III, FDP, and D-D are within the range of the seven coagulation factor composite quality control products (level 1), meeting the requirements of normal quality control products.

[0092] Example 2

[0093] This example provides the preparation of a seven coagulation factor composite quality control product (level 2). The difference from Example 1 is that in 200 ml of R1, add 300 ml of the diluent, 10 ml of the FIB adjustment solution, 30 ml of R0, 80 ml of PP1, mix well, add 50 μl of the FDP stock solution, 400 μl of the DD stock solution, and mix well. The component contents are shown in Table 2 below:

[0094] Table 2

[0095] Raw material R1 Diluent FIB Adjusting Solution R0 PP1 FDP DD Dosage 200ml 300ml 10ml 30ml 80ml 50 μl 400 μl Proportion 32% 48.4% 1.6% 4.8% 12.9% 0.008% 0.064%

[0096] After dispensing and freeze-drying, reconstitute with water and then detect. The results are shown in Table 3.

[0097] Table 3 Detection Results of Examples 1 and 2

[0098]

[0099] The results showed that APTT, PT, TT, FIB, AT-III, FDP, and D-D were within the range of the seven coagulation composite quality control products (level 2), meeting the requirements of abnormal quality control products.

[0100] Example 3

[0101] This example provides the preparation of a seven coagulation composite quality control product (level 2). The difference from Example 1 is that in 200 ml of R0, 110 ml of adsorbed human plasma, 25 ml of FIB adjusting solution 1, and 15 ml of PP0 were mixed evenly. Then 16.5 μl of FDP stock solution and 5.2 μl of DD stock solution were added and mixed evenly, and 850 μl of AT-Ⅲ was added. The components are shown in Table 4 below:

[0102] Table 4

[0103] R0 R1 FIB Adjusting Solution PP0 FDP DD AT-Ⅲ 200ml 110ml 30ml 16ml 16.5 μl 5.2 μl 850 μl 56% 30.8% 7% 4.48% 0.0046% 0.0015% 0.24%

[0104] Example 4

[0105] This example provides the preparation of a seven coagulation composite quality control product (level 2). The difference from Example 1 is that in 200 ml of R1, 330 ml of diluent, 13 ml of FIB adjusting solution, 34 ml of R0, and 85 ml of PP1 were mixed evenly. Then 50 μl of FDP stock solution and 400 μl of DD stock solution were added and mixed evenly. The components are shown in Table 5 below:

[0106] Table 5

[0107] R1 Diluent FIB Adjusting Solution R0 PP1 FDP DD 200ml 330ml 13ml 34ml 85ml 50 μl 400 μl 30.2% 49.8% 1.96% 5.13% 12.8% 0.0075% 0.06%

[0108] After the products of Example 3 and 4 were aliquoted, freeze-dried, reconstituted with water and then detected, the results are shown in Table 6:

[0109] Table 6 Detection Results of Example 3 and 4

[0110]

[0111] Comparing Example 1 and 3, through the above mixing of different raw material ratios, the values of indicators such as APTT, PT, TT, and FIB within the normal range can be adjusted satisfactorily. Comparing Example 2 and 4, through the above mixing of different raw material ratios, the values of indicators such as APTT, PT, TT, and FIB within the abnormal value range can be adjusted satisfactorily.

[0112] The above diluent can be one or several of D-trehalose, sodium chloride, HEPES FREE ACID, and trisodium citrate dihydrate. Among them, the percentage concentration range of D-trehalose is 0 to 100 g / L, the percentage concentration range of sodium chloride is 0 to 50 g / L; the percentage concentration range of HEPES FREE ACID is 0 to 30 g / L; the percentage concentration range of trisodium citrate dihydrate is 0 to 5 g / L, and the same applies hereinafter.

[0113] Experiment 1: Detection of the homogeneity of different samples.

[0114] The embodiments 1 and 2 of the present invention and commercially available quality control products (normal) (normal coagulation quality control product, abnormal coagulation quality control product (without TT), buffer matrix normal D-D quality control product, buffer matrix abnormal D-D quality control product, buffer matrix normal FDP quality control product, buffer matrix abnormal FDP quality control product) were accurately reconstituted with purified water according to the labeled volume, and the homogeneity test was carried out according to the following method:

[0115] Take 10 bottles of this product, and use the APTT, PT, TT, Fib, AT-Ⅲ, FDP, and D-Dimer detection kits to measure each bottle once (APTT, PT, TT, FIB, AT-Ⅲ, FDP, D-D) according to the respective instructions of the kits and the applicable coagulation analyzer instructions. Calculate the average value (1) of the test results according to formula 1 and the standard deviation (S1) of the test results according to formula 2; mix the above 10 bottles of reconstituted products and continuously test 10 times (APTT, PT, TT, FIB, AT-Ⅲ, FDP, D-D), calculate the average value (2) of the test results according to formula 1, calculate the standard deviation (S2) of the test results according to formula 2; calculate the within-bottle coefficient of variation CV within (APTT, PT, TT, FIB, AT-Ⅲ, FDP, D-D) according to formula 3; calculate the between-bottle coefficient of variation CV between (APTT, PT, TT, FIB, AT-Ⅲ, FDP, DD) according to formula 4 and formula 5.

[0116] Formula 1 ;

[0117] Formula 2 ;

[0118] Formula 3 ;

[0119] Formula 4 ;

[0120] Formula 5 (when S1 < S2, let CV between = 0);

[0121] Where: ----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 "General Requirements for Quality Control Substances for In Vitro Diagnostic Reagents" (YY / T 1652-2019). The results are shown in Table 7 below:

[0122] Table 7 Detection Results of the Homogeneity of Different Samples

[0123]

[0124] From the above results, it can be seen that for all items in Examples 1 and 2, both the between-bottle CV and within-bottle CV are less than those of the commercially available quality control products. For the between-bottle CV 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.

[0125] Experiment 2 Detection of the Stability of Different Samples.

[0126] The quality control products of Examples 1 and 2 of the present invention and the commercially available quality control products (normal) (normal coagulation quality control product, abnormal coagulation quality control product (without TT), buffer matrix normal D-D quality control product, buffer matrix abnormal D-D quality control product, buffer matrix normal FDP quality control product, buffer matrix abnormal FDP quality control product) were accurately reconstituted with purified water according to the labeled volume and stored under refrigerated conditions (2°C - 8°C). Samples were taken at 0h, 4h, 6h, 8h, 17h, 24h, and 48h to detect APTT, PT, TT, FIB, ATIII, D-D, and FDP. The stability deviation results were calculated with 0h as the reference and are shown in Tables 4 to 7.

[0127] (Stability deviation = ). The results are shown in Tables 8 - 11 below:

[0128] Table 8 Reconstitution Stability Deviation of the Quality Control Product of Example 1

[0129]

[0130] Table 9 Reconstitution Stability Deviation of the Quality Control Product of Example 2

[0131]

[0132] Table 10 Reconstitution Stability Deviation of the Commercially Available Normal Quality Control Product

[0133]

[0134] Table 11 Reconstitution Stability Deviation of the Commercially Available Abnormal Quality Control Product

[0135]

[0136] As can be seen from the results in Tables 8 - 11 above, the stability deviation results of all items in Examples 1 and 2 are less than those of the commercially available quality control products, proving that the stability of the quality control products of the present invention is superior to that of the commercially available quality control products.

[0137] Experiment 3: Taking Example 1 as an example, six control groups 1 - 6 were set up to study the influence of different contents of each component on the results.

[0138] For other contents, the same as in Example 1, the situations when setting different contents of each component are as follows:

[0139] 1. Control group 1 was set up, which differed from Example 1 in that the dosage of FDP stock solution 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.

[0140] Table 12 Dosage of FDP stock solution and FDP measurement value

[0141] FDP Dosage FDP (ug / ml) D-D (ug / ml) APTT (s) PT (s) TT (s) Fib (g / L) AT-Ⅲ (%) 0 μl 1.78 0.33 30.93 12.07 12.34 2.89 88.57 30 μl 5.21 0.38 30.86 11.95 15.98 2.91 88.46 50 μl 9.08 0.36 30.25 12.07 16.15 2.94 88.14 70 μl 12.43 0.39 31.25 12.25 16.17 2.92 88.32 100 μl 20 0.37 30.97 12.03 16.27 2.97 88.47

[0142] The results in Table 12 show that, with the dosage of other components remaining unchanged, as the dosage of FDP stock solution increased within the range of 0 - 100 μl, the FDP test value increased accordingly, and the changes in the other six test values were not obvious.

[0143] 2. Control group 2 was set up, which differed from Example 1 in that the dosage of DD stock solution 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.

[0144] Table 13 Dosage of D - D stock solution and D - D measurement value

[0145] DD Dosage D-D (ug / ml) FDP (ug / ml) APTT (s) PT (s) TT (s) Fib (g / L) AT-Ⅲ (%) 0 μl 0.22 3.01 30.86 12.21 16.26 2.98 88.31 10 μl 0.43 2.96 30.76 12.17 16.17 2.95 88.37 30 μl 0.84 3.00 30.94 12.18 16.21 2.93 89.06 70 μl 1.74 2.95 30.84 12.09 16.24 2.91 88.59 100 μl 2.45 2.97 30.79 12.19 16.23 2.94 88.34

[0146] The results in Table 13 show that, with the dosage of other components remaining unchanged, as the dosage of DD stock solution increased within the range of 0 - 100 μl, the DD test value increased accordingly, and the changes in the other six test values were not obvious.

[0147] 3. Control group 3 was set up, which differed from Example 1 in that the dosage of AT - III concentrate 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.

[0148] Table 14 Dosage of AT - III concentrate and AT - III measurement value

[0149] AT-Ⅲ Dosage AT-Ⅲ (%) D-D (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 2.94 300 μl 82.42 0.32 3.01 30.79 12.13 16.23 2.95 900 μl 87.65 0.33 2.94 30.83 12.15 16.13 2.97 1500 μl 92.10 0.32 3.03 30.87 12.18 16.26 2.93 2100 μl 96.94 0.33 2.99 30.46 12.19 16.24 2.96 2700 μl 101.77 0.34 2.99 30.52 12.20 16.27 2.93

[0150] As can be seen from the results in Table 14, when the concentrate was not added, the AT-III activity was about 80% of that of normal people. When the dosages of other components remained unchanged, as the dosage of AT-III concentrate increased within the range of 0 - 2700 μl, the detected value of AT-III increased correspondingly, while the detected values of the other six items showed no obvious changes.

[0151] 4. Set control group 5, which is different from Example 1 in that the AT-III concentrate, FDP stock solution, and DD stock solution are not added, and the concentration of the 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.

[0152] Table 15 Influence of the change in the concentration of the FIB regulating solution on the coagulation activity

[0153] FIB Adjusting 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

[0154] As can be seen from the results in Table 15, when the dosages of other components remained unchanged, with the volume of the FIB regulating solution remaining the same and the concentration of the FIB regulating solution increasing within the range of 0 - 130 g / dL, the detected value of FIB increased, and the measured values of APTT, PT, and TT gradually shortened, while the change in the measured value of AT-III was not significant.

[0155] 5. Set control group 5, which is different from Example 1 in that the AT-III concentrate, FDP stock solution, and DD stock solution are not added, and the dosage of PP0 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.

[0156] Table 16 Influence of the change in the dosage of PP0 on the coagulation activity

[0157] 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

[0158] As can be seen from the results in Table 16, when the dosages of other components remained unchanged, as the dosage of PP0 increased within the range of 0 - 40 mL, the detected values of APTT and PT shortened accordingly, the detected value of TT increased slightly, and the measured values of FIB and AT-III showed no obvious changes.

[0159] 6. Set control group 6, which is different from Example 1 in that the AT-III concentrate, FDP stock solution, and DD stock solution are not added, the total amount of R0 and R1 remains unchanged, and their ratio is changed. The detection results are as follows in Table 17:

[0160] Table 17 Influence of the change in the volume ratio of R0 to R1 on the coagulation activity

[0161] 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

[0162] The results in Table 17 show that when the dosages of other components remain unchanged, the total volume of the dosages of R0 and R1 remains unchanged, and as the dosage of R0 decreases while the dosage of R1 gradually increases, the APTT and PT test values increase accordingly, while the TT, FIB, and AT-III test values show no obvious changes.

[0163] Experiment 4: Taking Example 2 as an example, set up Control Groups 7 - 13 to study the influence of different contents of each component on the results

[0164] For other contents, refer to those in Example 2. The situations when setting different contents of each component are as follows:

[0165] 1. Set up Control Group 7, which is different from Example 2 in that the dosage of the FDP stock solution changes from 50 μl to 0 μl, 25 μl, 50 μl, 75 μl, and 100 μl. The test results are shown in Table 18 below.

[0166] Table 18 Dosage of FDP stock solution and FDP measurement value

[0167] FDP Dosage FDP (ug / ml) D-D (ug / ml) APTT (s) PT (s) TT (s) Fib (g / L) AT-Ⅲ (%) 0 μl 1.04 2.61 57.33 39.70 31.35 1.42 38.71 25 μl 10.25 2.59 57.45 39.76 31.42 1.41 38.74 50 μl 18.06 2.62 57.48 39.71 31.49 1.44 38.69 75 μl 26.32 2.58 57.43 39.61 31.36 1.43 38.62 100 μl 35.52 2.63 57.49 37.82 31.47 1.43 38.65

[0168] The results in Table 18 show that when the dosages of other components remain unchanged, as the dosage of the FDP stock solution increases within the range of 0 - 100 μl, the FDP test value increases accordingly, and the other six test values show no obvious changes.

[0169] 2. Set up Control Group 8, which is different from Example 2 in that the dosage of the DD stock solution changes from 400 μl to 0 μl, 200 μl, 700 μl, and 1000 μl. The test results are shown in Table 19 below.

[0170] Table 19 Dosage of D - D stock solution and D - D measurement value

[0171] DD Dosage D-D (ug / ml) FDP (ug / ml) APTT (s) PT (s) TT (s) Fib (g / L) AT-Ⅲ (%) 0 μl 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 38.72 400 μl 2.62 18.06 57.48 39.71 31.49 1.44 38.69 700 μl 4.94 18.10 57.51 39.68 31.45 1.45 38.64 1000 μl 6.48 18.07 57.46 37.78 31.49 1.46 38.62

[0172] The results in Table 19 show that when the dosages of other components remain unchanged, as the dosage of the DD stock solution increases within the range of 0 - 1000 μl, the DD test value increases accordingly, and the other six test values show no obvious changes.

[0173] 3. Set up Control Group 9, which is different from Example 2 in that no FDP stock solution and DD stock solution are added, and the dosage of the FIB adjusting solution changes from 10 mL to 0 mL, 5 mL, 20 mL, 30 mL, and 40 ml. The results are shown in Table 20 below.

[0174] Table 20 Influence of the change in the dosage of the FIB adjusting solution on the coagulation activity

[0175] FIB Adjusting Solution Volume Fib (g / L) APTT (s) PT (s) TT (s) AT-Ⅲ (%) 0 mL 0.61 89.64 61.28 49.26 38.26 5 mL 1.03 68.43 50.62 39.47 38.47 10 mL 1.44 57.48 39.71 31.49 38.69 20 mL 2.25 46.82 32.59 24.51 38.79 30 mL 3.09 41.19 27.28 19.43 38.94 40 mL 3.95 39.49 21.57 17.31 39.12

[0176] As can be seen from the results in Table 20, when the dosages of other components remain unchanged, within the range of 0 - 40 mL of the FIB conditioning solution dosage, the FIB detection value increases accordingly, and the measured values of APTT, PT, and TT gradually shorten, while the measured value of AT-III does not change significantly.

[0177] 4. Set up Control Group 10, which is different from Example 2 in that the FDP stock solution and DD stock solution are not added, and the dosage of PP1 changes from 80 mL to 0 mL, 40 mL, 120 mL, and 160 mL. The results are shown in Table 21 below.

[0178] Table 21 Effects of Changes in PP1 Dosage on Coagulation Activity

[0179] 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 40 mL 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

[0180] As can be seen from the results in Table 21, when the dosages of other components remain unchanged, within the range of 0 - 160 mL of the PP1 dosage, the measured values of APTT and PT shorten accordingly, the measured value of TT is slightly prolonged, and the measured values of FIB and AT-III do not change significantly.

[0181] 5. Set up Control Group 11, which is different from Example 2 in that the FDP stock solution and DD stock solution are not added, and the dosage of R0 changes from 30 mL to 0 mL, 15 mL, 50 mL, and 70 mL. The results are shown in Table 22 below.

[0182] Table 22 Effects of Changes in R0 Dosage on Coagulation Activity

[0183] 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 15 mL 60.24 41.13 31.47 1.41 37.02 30 mL 57.48 39.71 31.49 1.44 38.69 50 mL 52.43 37.16 31.51 1.46 39.51 70 mL 48.27 35.28 31.43 1.49 40.26

[0184] The results in Table 22 show that when the dosages of other components remain unchanged, within the range of 0 - 70 mL of the R0 dosage, the measured values of APTT and PT decrease correspondingly, and the measured values of TT, FIB, and AT-III do not change significantly.

[0185] 6. Set up Control Group 12, which is different from Example 2 in that the FDP stock solution and DD stock solution are not added, the total amount of R1 and the diluent remains unchanged, and their ratio changes. The detection results are as follows in Table 23:

[0186] Table 23 Effects of Changes in the Volume Ratio of R1 to the Diluent on Coagulation Activity

[0187] R1 Dosage Diluent Dosage 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

[0188] The results in Table 23 show that when the dosages of other components remain unchanged, the total volume of the dosages of R1 and the diluent remains unchanged, as the dosage of R1 increases and then gradually decreases, the measured values of APTT, PT, and TT shorten correspondingly, and the measured values of FIB and AT-III increase gradually.

[0189] The above embodiments / test examples are merely illustrations for clear explanation and are not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or alterations can be made based on the above description. It is not necessary and impossible to enumerate all implementation manners here. And the obvious changes or alterations derived therefrom still fall within the protection scope of the present invention.

Claims

1. A preparation method of a seven - item combined coagulation quality control product, characterized in that: Comprising the following steps: (1) Obtain human mixed plasma anticoagulated with sodium citrate and animal plasma PP anticoagulated with sodium citrate; the animal is one of cattle, pigs, sheep, and horses; (2) Part of the human mixed plasma and PP are respectively adsorbed by aluminum hydroxide colloid, centrifuged to obtain the supernatant, and adsorbed human mixed plasma and adsorbed PP are obtained; (3) Another part of the human mixed plasma and PP, as well as the adsorbed human mixed plasma and adsorbed PP, are respectively added with a buffer stabilizing protective agent to obtain corresponding R0, PP0, R1, and PP1; (4) Divide R0 and R1 into two parts: Add PP0, the first part of R1, and FIB adjusting solution to the first part of R0, and adjust the FIB, APTT, PT, and TT values to be consistent with those of normal human mixed plasma to obtain level 1 matrix solution 1, where the target values are APTT of 21 - 45 sec, PT of 10 - 15 sec, TT of 10 - 20 sec, and FIB of 2 - 4 mg / mL; Add PP1, the second part of R0, diluent, and FIB adjusting solution to the second part of R1, and adjust the APTT, PT, TT, FIB, and AT-Ⅲ values to the target values to obtain level 2 matrix solution 1, where the target values are APTT of 45 - 150 sec, PT of 15 - 74 sec, TT of 20 - 50 sec, FIB of 0.6 - 2 mg / mL, and AT-Ⅲ of 25 - 80%; (5) Add DD stock solution and FDP stock solution to level 1 matrix solution 1, and adjust the DD and FDP concentrations in level 1 matrix solution 1 to the target values to obtain level 1 matrix solution 2, where DD is 0.1 - 0.9 μg / mL and FDP is 2.2 - 5.5 μg / mL; then add AT-Ⅲ concentrate to level 1 matrix solution 2 and adjust the AT-Ⅲ value to be consistent with that of normal human mixed plasma, and thus obtain the coagulation seven-item composite quality control product level 1, where the target value of AT-Ⅲ is 80 - 120%; Add DD stock solution and FDP stock solution to level 2 matrix solution 1, and adjust the DD and FDP concentrations 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 are DD of 0.9 - 30 μg / mL and FDP of 5.5 - 55 μg / mL; Subpackage and lyophilize to obtain two-level activated coagulation detection reagent quality control products.

2. The preparation method of a coagulation seven-item composite quality control product according to claim 1, wherein: In step (2), the dosage of aluminum hydroxide colloid is 1% - 10% (V / V).

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

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

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

6. The preparation method of a seven - item coagulation combined quality control product according to claim 1, characterized in that: In the step (5), for the coagulation factor VII composite quality control product at level 2, the prothrombin time is 15 - 74 s, the activated partial thromboplastin time is 45 - 150 s, the fibrinogen concentration is 0.6 - 2 g / L, the antithrombin III activity is 25 - 80%, the D-dimer concentration is 0.9 - 3.0 μg / mL, the fibrin degradation product concentration is 5.5 - 55 μg / mL, and the thrombin time is 20 - 50 s.

7. A coagulation factor VII composite quality control product prepared by the method according to any one of claims 1 - 6.

8. A seven - item coagulation combined quality control product according to claim 7, characterized in that: The target target values of the coagulation factor VII composite quality control product at level 1 are: APTT 25 - 31 s, PT 10.8 - 13 s, TT 14 - 18 s, FIB 2.2 - 3.2 g / L, AT-Ⅲ 86 - 105%, D-D 0.3 - 0.5 ug / ml, FDP 2.5 - 5 ug / ml.

9. The coagulation seven - item composite quality control product according to claim 8, wherein: The target target values of the coagulation factor VII composite quality control product at level 2 are: APTT 48 - 65 s, PT 28 - 36 s, TT 28 - 50 s, FIB 1.2 - 1.7 g / L, AT-Ⅲ 30 - 50%, D-D 2.0 - 3.0 ug / ml, FDP 15 - 20 ug / ml.

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