| To detect the effect of quantity and platelet concentration of platelet-rich plasma(PRP),mixing times,concentration and storage time of inducer,incubation time,temperature,anticoagulants and extracellular Ca2+ concentrations on test results of platelet aggregation(PA),thromboelastography(TEG) and platelet membrane glycoproteins. So that to provide testing evidences for establishing Total Quality Control system of platelet function tests.Blood samples of 519 outpatient medical volunteers were collected, then under different factors mentioned above, the rate of platelet aggregation (n=394), parameters of TEG (n=94) and platelet membrane glycoproteins(n=31) were detected according to the standard protocols by plasma turbidimetry, recalcification and flow cytometry respectively.Platelet aggregation is a analysis means effected by many factors.When quantity of platelet-rich plasma (PRP) is 300ul, AA and ADP store for 7 months and 13 months in a-50℃refrigeration and AA and ADP after thawing store for 24h at 5℃respectively, the ffect on the testing result of platelet aggregation is not significiant; The platelet concentration of 250×109/L seems to be the optimal concentration for platelet aggregation by plasma turbidimetry. The AA and ADP concentration of 0.7mmol/L and lOumol/L seems to be the optimal concentration at platelet concentration of 250×109/L. When the platele concentration is in the range of 180～400×109/L,the effect on the testing result of platelet aggregation is not significant. Decreasion of platelet aggregation as time elapse may be caused by aging of sample and a high PH. After blood ample drawn,the test of platelet aggregation induced by ADP should be finished in 5h. And the test induced by AA concentration of 0.5mmol/L and 1.0mmol/L should be done in 4h and 8h respectively; The rate of platelet aggregation decreases with a lower Ca2+ concentration when Ca2+ concentration is in the range of 0.04～0.11mmol/L and increases with a higher Ca2+ when Ca2+ is in the range of 0.1～33.7mmol/L. The rate indued by ADP decreases markedly at the concentration of 0.08 mmol/L;A high calcium (≥39 mmol/L) can inhibit the platelet aggregation; The effect of different anticoagulatants on platelet aggregation and glycoproteins activation is significant, sodium citrate seems to be the best anticoagulant for platelets aggregation. Mixing times of blood samle in kaolin tube should be fixed at 3-5 times in the test of TEG, or the testing result of R and MA of CK are not incomparable; Blood sample for TEG test is stored in a low temperature site, which can enhance the activity of blood coagulation factors,fibrinogen and platelet function activated by AA and ADP; Sample is stored in a high temperature site, which can enhance the activity of blood coagulation factors and reduce activity of platelet activated by ADP; The effect of sample at room temperature for 8h on the parameter of RCK,KCFK,αCK,MACK,KA,MAA,and MAAA is not significant, the activity of platelet activated by ADP (MAADP) reduces as time elapses;So blood sample after drawn for TEG test shoud be stored at room tempreture and be finished in 1-2h. Calcium citrate itself may impair coagulation dynamics-blood coagulation factors and platelet. Coagulation analyses using blood that has been exposed to citrate and recalcified do not yield reliable depictions of the natural dynamics of blood coagulation processes; The Ca2+ concentration of 2.1 mmol/L seems to be the optimal concentration for thromboelastography by recalcification method. The effect of different anticoagulatants on glycoproteins activation is significant, CTAD has less effect on self-activated platelet in flow cytometry and seems to be the best specimen anticoagulating methed. |
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