血小板測(cè)試方法學(xué)

1、ADVIA 2120/120 二維血小板檢測(cè),背景介紹,常規(guī)自動(dòng)化 PLT 檢測(cè)（一維）,檢測(cè)原理：利用血小板體積單一參數(shù)來檢測(cè)和計(jì)數(shù)檢測(cè)技術(shù)：電阻法 (Coulter 原理)光學(xué)法PLT體積檢測(cè)范圍：Sysmex1 - 40 fLCoulter2 - 20 fLCell－Dyn 1 - 35 fLH·Systems1 - 20 fL （Optical）局限性：僅依靠P

2、LT 體積參數(shù)排除干擾因素，無法準(zhǔn)確區(qū)分： Microcytes 、RBC fragment、RBC ghost、Large PLT,Abbott CellDyn 4000 DataOptical PLT 180 *Impedance PLT 248 ** difference due to Microcytes,電阻抗法,光學(xué)法,常規(guī) PLT 檢測(cè)技術(shù),存在不足,血小板檢測(cè)面臨的挑戰(zhàn),RBC fragments

3、WBC fragments 和 complexes細(xì)菌、寄生蟲細(xì)胞碎片 、雜質(zhì)大血小板 、血小板微泡 交叉污染血小板聚集抗凝集影響,測(cè)試原理,ADVIA 2120 2D－PLT檢測(cè),二維參數(shù)：PLT體積 (V) 和 折射指數(shù) (MPC) 采用專利的Mie原理進(jìn)行激光散射分析對(duì)血小板細(xì)胞逐一進(jìn)行二維檢測(cè)提高對(duì)血小板、 細(xì)胞碎片、低色素細(xì)胞和小紅細(xì)胞的鑒別PLT體積檢測(cè)范圍：1 - 60 fL血小板計(jì)數(shù)包 括大血小板

4、（>20fL）提高對(duì)化療、免疫性血小板減少癥（ITP）、 Bernard-Soulier綜合癥、骨髓增值癥等大血小板樣本計(jì)數(shù)的準(zhǔn)確性,,,,,,RBC/PLT試劑,細(xì)胞球形固定,RBC,Platelets,,,參考文獻(xiàn)：Tycko PH，Met MH,Epstein EA, Grinbaum:紅細(xì)胞體積與血紅蛋白的流式細(xì)胞光散射測(cè)定，應(yīng)用光學(xué)24（9）：1355-1365（1985）,流式細(xì)胞技術(shù)PLT檢測(cè)原理,流式細(xì)胞技術(shù)P

5、LT檢測(cè)原理,線性范圍：5 – 3500 X 103/µL體積范圍： 1 - 60 fL包括大血小板 >20fL（RI: 1.35~1.40）排除干擾：RBC fragment、ghost、 microcytes,2D－PLT 檢測(cè)細(xì)胞散射圖,高角度 (5º to 15 º),低角度 (2º to 3 º),參數(shù):Large PLT

6、MPCMPMPCDWP Count-2DPCTPDWPLTNPLTXPLTYPMDWR Count-2DRBC-2D CountRBC FragmentsRBC Ghosts,解釋:大血小板計(jì)數(shù)平均血小板成分濃度平均血小板聚集血小析成分分布寬度在血小板測(cè)定中血小板的原始計(jì)數(shù)血小板壓積血小板體積平均分布寬度血小板折射率平均值血小板高角度信號(hào)(放大)平均值血小板低角度信號(hào)(放大)平均值

7、在血小板測(cè)定中紅細(xì)胞原始計(jì)數(shù)在血小板測(cè)定中紅細(xì)胞計(jì)數(shù)紅細(xì)胞碎片計(jì)數(shù)紅細(xì)胞影計(jì)數(shù),ADVIA 2120/120 PLT檢測(cè)參數(shù),15 項(xiàng)研究參數(shù)（供實(shí)驗(yàn)室、臨床研究）,2 項(xiàng)報(bào)告參數(shù)：PLT 和 MPV,測(cè)試表現(xiàn),ADVIA 120 2D－PLT與電阻抗法 和免疫法檢測(cè) 比較研究,三種不同PLT檢測(cè)方法:CD61單克隆抗體免疫法 PLT計(jì)數(shù)（公認(rèn)參考方法）電阻抗計(jì)數(shù)法ADVIA 120 2D－PLT檢測(cè)法Study by

8、 Dr. Samuel Machin, University College, LondonResults published in the British Journal of Haematology 2000 108:2, 228-235,Discrepant Samples in Machin Study,One-dimensional PLT counts VS Reference manual counts,J. Kunic

9、ka, G. Fischer, J. Murphy, D. Zelmanovic. “Improved platelet counting using two-dimensional laser light scatter”. AJCP (2000), in press.,two-dimensional platelet counts VS Reference manual counts,J. Kunicka, G. Fischer,

10、J. Murphy, D. Zelmanovic. “Improved platelet counting using two-dimensional laser light scatter”. AJCP (2000), in press.,two-dimensional platelet and one-dimensional PLT counts VSthe reference manual counts,J. Kunicka

11、, G. Fischer, J. Murphy, D. Zelmanovic. “Improved platelet counting using two-dimensional laser light scatter”. AJCP (2000), in press.,Relationship between the RBC fragment count and the difference between the one- and

12、two-dimensional PLT counts in 260 samples,J. Kunicka, G. Fischer, J. Murphy, D. Zelmanovic. “Improved platelet counting using two-dimensional laser light scatter”. AJCP (2000), in press.,兩類PLT 檢測(cè)方法比較,ADVIA 2120/120二維參數(shù)

13、PLT體積和折射指數(shù)（RI）線性范圍寬：5－3500，適用于極高和極低PLT值樣本準(zhǔn)確區(qū)分并排除microcytes, RBC fragments 和 RBC ghosts對(duì) PLT 計(jì)數(shù)的干擾潛在的臨床應(yīng)用價(jià)值的參數(shù) ： MPC和MPM,常規(guī)檢測(cè)儀器一維參數(shù)僅PLT 體積（V）線性范圍窄：0－2000，尤其對(duì)低PLT值樣品差干擾顆粒可能被誤計(jì)入血小板總數(shù)，造成PLT過高；或因大血小板漏計(jì)而PLT過低,臨床應(yīng)用,ADVIA

14、 2120/120血小板檢測(cè)臨床表現(xiàn)和臨床意義,高值 PLT 計(jì)數(shù)：對(duì)血小板增多癥患者降細(xì)胞治療的監(jiān)控低值 PLT 計(jì)數(shù)：提示血小板輸血的適應(yīng)癥對(duì)患者化療全程監(jiān)控 對(duì)先天性、免疫性和非免疫性血小板減少癥的檢測(cè)、監(jiān)控對(duì)提高血小板的藥物治療的監(jiān)控移植后的骨髓恢復(fù)情況的監(jiān)控,理想血球分析儀：更精確、更廣的血小板檢測(cè)能力,Bernard Soulier 綜合癥,ADVIA 120PLT = 68,000 / uL電阻法：

15、PLT = 24,000 / uL,動(dòng)物－貓的 RBC 和 PLT 檢測(cè),貓的血液中含有非常大的PLT和非常小的RBCRBC: 8.2 x 10 12/LMCV: 44.8 fLPLT: 447 x 10 9 /LMPV: 17.1 fL,大血小板、小紅細(xì)胞病例Case #503,ADVIA 2120 2D - PLT 檢測(cè)：PLT 49,000Large PLT 16,000MPV 15.8 fLADVI

16、A 2120 報(bào)警提示：MICRO (+++), Large Platelets (+++)手工 PLT 計(jì)數(shù) 47,00030% Large Platelets, Microcytes,血小板減少癥（ RBC fragments） Case #58642,ADVIA 2120 2D – PLT檢測(cè)：10 x 103 cells/uL10 x 103 RBC Fragments手工計(jì)數(shù)：8 x 103 cells/u

17、L一維光學(xué)法檢測(cè)：22 x 103 cells/uL,異形紅細(xì)胞癥和小紅細(xì)胞癥 病例Case #304,ADVIA 2120 2D - PLT 檢測(cè)： PLT 144,000RDW 28.5%MPV 10.7 fLADVIA 2120 報(bào)警提示：MICRO (+++), HYPO (+++), ANISO (+++)手工法 PLT 計(jì)數(shù)：PLT 117,000,優(yōu)勢(shì)總結(jié),ADVIA 2120/1

18、20 2D-PLT檢測(cè)總結(jié),ADVIA 2120/120 是唯一真正采用二維方法（體積和折射指數(shù)）實(shí)現(xiàn)對(duì)PLT 準(zhǔn)確檢測(cè)的血球分析儀 無論正常還是干擾因素的樣品, ADVIA 2120/120在血小板檢測(cè)的精密度及準(zhǔn)確性方面與參考方法以及形態(tài)學(xué)觀察都有很好的吻合度 ADVIA 2120/120 2D-PLT 檢測(cè)技術(shù)明顯優(yōu)于電阻抗法和光學(xué)法，尤其在血小板疾病以及異常干擾的臨床病例,Selected References,Improv

19、ed Platelet Counting: Multi-Dimensional Analysis of Platelet Light Scatter. J. Kunicka, G. Fischer, J. Murphy, D. Zelmanovic. Amer. J. Clin. Path. 2000 (In Press).Advances in Routine Laboratory Platelet Counting. S.J.Ma

20、chin, P.Harrison, D. Grant and C.Briggs. British Journal of Haematology 2000 108:2, 228-235Use of Mean Platelet Component to Measure Platelet Activation on the ADVIA 120 Haematology System. M.G. Macey, E. Carty, L. Webb

21、, E.S. Chapman, D. Zelmanovic, D. Okrongly, D.S. Rampton, A.C. Newland. Cytometry 38, 250 2019Stability Of Various Anticoagulated Whole Blood Specimens For Use In Ex Vivo Platelet Activation Measurements. E. S. Chapman,

22、 G. Kling, W. Canfield, D. Zelmanovic. Blood 2019 94:63B, 10, Suppl. 1 Abstract # 3437.Monitoring In Vitro And Ex Vivo Platelet Activity: Comparison Of Alpha Granule Release, Density Distribution, Platelet Adhesion And