Buffer solution for detecting blood coagulation factor VIII inhibitor and detection method
A technology for coagulation factors and inhibitors, which is applied in the field of buffers for the detection of coagulation factor VIII inhibitors, can solve the problems of complicated operation procedures, difficult to grasp operation specifications and quality control, and achieve the effect of improving stability and good buffering capacity.
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Embodiment 1
[0041] The buffer solution used in the detection of blood coagulation factor VIII inhibitors provided in this example contains, per L, 2.72g imidazole, 4.68g NaCl, 8.4g BSA, 0.02ml Tween-20, 6.5g NaN 3 and 14.88 mmol HCl.
[0042] The present embodiment provides the preparation method of above-mentioned buffer solution, as follows:
[0043] 1. Weigh 2.72g imidazole, 4.68g NaCl, 8.4g BSA, 0.02ml Tween-20, and 6.5g NaN 3 Place in a volumetric flask, add 500ml of distilled water, stir to dissolve;
[0044] 2. Add 148.8ml 0.1M hydrochloric acid, mix well and adjust the pH to 7.3;
[0045] 3. Dilute to 1L with distilled water and adjust the pH to 7.3.
[0046]The buffer used in the detection of coagulation factor VIII inhibitors provided in this example has good buffering capacity and can be used to dilute blood samples during the detection of coagulation factor VIII or coagulation factor VIII inhibitors to improve the stability of detection results sex.
Embodiment 2
[0048] The method for detecting coagulation factor VIII provided by the present embodiment comprises the steps of:
[0049] 1 Sample Collection
[0050] Blood was collected from the elbow veins of 20 healthy normal subjects (samples were numbered 1, 2, 3..., 20). Put on a tourniquet before the start of blood collection, use 109mmlol / L sodium citrate anticoagulant (blood: sodium citrate=9:1) vacuum blood collection tube to collect 3ml of blood sample, note that after the blood is pumped into the anticoagulant tube, it is necessary to Mix by gently inverting five times.
[0051] This step is optional.
[0052] 2 Separation of plasma
[0053] Freshly collected blood samples were separated from the plasma within 1 hour, and the blood collection tube was placed in a centrifuge, centrifuged at a centrifugal force of 1600g for 15 minutes, and the plasma was transferred to a 1.5ml centrifuge tube with a 200μl micropipette, and the plasma was stored at room temperature It can be st...
Embodiment 3
[0067] The method for the detection of blood coagulation factor VIII inhibitor that the present embodiment provides, concrete operation is as follows:
[0068] 1 Detection of residual coagulation factor VIII activity
[0069] 1.1 Using the imidazole buffer solution provided in Example 1 as the plasma diluent, in a plastic test tube, the blood plasma of a hemophilia patient who has produced coagulation factor VIII antibodies due to long-term use of recombinant coagulation factor VIII drug treatment (after testing, the patient's plasma The blood coagulation factor VIII activity is 0.001%) carry out doubling dilution (1 times, 5 times, 10 times, 20 times, 40 times, 80 times, 160 times, 320 times), the plasma volume of each multiple is 0.2ml.
[0070] 1.2 Inhale 0.2ml of coagulation factor VIII-deficient plasma into another plastic test tube as a standard plasma control.
[0071] 1.3 Add 0.2ml of normal mixed plasma into the standard tube and the diluted plasma to be tested. At ...
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