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Rapid flow cytometric quantitation of reticulated platelets in whole blood

 

作者: ChavdaN.,   MackieI. J.,   PorterJ. B.,   HarrisonP.,   PattersonK.,   MachinS. J.,  

 

期刊: Platelets  (Taylor Available online 1996)
卷期: Volume 7, issue 4  

页码: 189-194

 

ISSN:0953-7104

 

年代: 1996

 

DOI:10.3109/09537109609023578

 

出版商: Taylor&Francis

 

数据来源: Taylor

 

摘要:

Young or reticulated platelets contain some residual mRNA, which is rapidly degraded after platelet release into the circulation. In order to minimize platelet activation and possible loss of large platelets during sample handling a whole blood method has been developed utilising the RNA fluorochrome thiazole orange (TO) in combination with an antibody to anti-glycoprotein Ib (GpIb) directly conjugated to phycoerythrin (PE), to specifically stain reticulated platelets via flow cytometric analysis. In this study whole blood analysis of platelet mRNA was undertaken in healthy normal subjects and a variety of patients with haematological abnormalities. The percentage of Gp Ib positive platelets containing mRNA in normals (n= 22) was 11.61% with a two SD range of 3.19-20.01%. The percentage of reticulated platelets was significantly increased (mean mRNA content±one SD) in sickle cell disorders (n= 22) 38.12%±18.42 (P<0.001); thalassaemia (major, intermedia and trait) (n= 24) 29.76±19.15 (P<0.001); ITP (N= 20) 23.53%±13.04 (P<0.02) and essentialthrombocythemia (N= 32) 37.12±19.84 (P<0.001). Platelets from patients with reactive thrombocytosis (N= 15) were only 12.23% positive (±6.95) and not significantly different from the normal range (P= 0.95). This method offers a rapid and simple procedure for assessment of reticulated platelets in whole blood and suggests that there may be an increased platelet turnover in certain haemoglobinopathies.

 

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