Thrombin solution
A technology of thrombin and solution, which is applied in the field of enzyme industry and can solve the problems of inconvenient preservation of thrombin solution
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Embodiment 1
[0054] Preparation of thrombin time (TT) detection reagent containing stabilizer of the present invention
[0055] Weigh 11.9g of HEPES acid, 7.7g of calcium lactate, 50g of sodium glutamate, 1g of sodium azide, and 0.2g of Tween 80, dissolve in distilled water, adjust the pH to 6.4 with sodium hydroxide, and dilute to 1L. Take 4 pieces of thrombin (1000U / cart), dissolve it with 1mL / cartridge of the above preparation buffer, take 3.6mL of dissolved thrombin (the amount of enzyme can be adjusted to control the difference between batches), add it to 1L buffer solution and shake well, and prepare the TT detection reagent Finish.
[0056] Product performance index
[0057] 1. Repeatability: coefficient of variation (CV) ≤ 5%; repeat the test 10 times with normal quality control plasma, and calculate the average value of the determination and standard deviation(s). Calculate the coefficient of variation (CV) according to formula (1). The obtained results should meet the follow...
Embodiment 2
[0066] Detection method of thrombin time (TT) detection reagent
[0067] Take 100 μL of plasma, preheat at 37°C for 2 minutes, then add 100 μL of thrombin reagent, and measure in CA550 automatic coagulation instrument (manufacturer SYSMEX) according to the manufacturer's instructions.
[0068] Precautions:
[0069] 1. Blood collection should avoid hemolysis and prevent interstitial fluid from mixing. The blood draw should be smooth, the anticoagulation should be sufficient, and there should be no clots; the platelets must be removed when separating the plasma.
[0070] 2. EDTA and heparin should not be used as anticoagulant during sample collection. The ratio of blood and anticoagulant should be accurate, and specimens with deviation of blood volume greater than 10% should not be used.
[0071] 3. If the hematocrit is ≤20% or ≥55%, the ratio of blood sample to anticoagulant needs to be adjusted, and the number of milliliters of anticoagulant=0.00185×the number of milliliter...
Embodiment 3
[0078] Stability experiment of the thrombin time (TT) detection reagent containing stabilizer of the present invention
[0079] Prepare thrombin time (TT) detection reagent according to the scheme of embodiment 1, use CA550 automatic coagulation instrument (manufacturer SYSMEX) to measure the stability of the reagent described under the following conditions according to the manufacturer's instructions
[0080] 1. Accelerated destruction at 37°C.
[0081] 2. 2-8 ℃ open bottle stability.
[0082] 3. Long-term stability at 2-8°C.
[0083] Stable and accelerated destruction at 37°C, the measured value of normal quality control plasma, the measured value is stable within 1 month. See Table 1.
[0084] Table 1 Accelerated thermal damage test at 37℃
[0085]
[0086] Stability test after opening the bottle, put it at 2-8°C for 15 days after opening the bottle, the measured value is relatively stable. See Table 2.
[0087] Table 2 2-8 ℃ bottle opening stability
[0088]
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