Usefulness of Fully Automated Measurement of Reticulated Platelets Using Whole Blood

Abe, Yasunori; Wada, Hideo; Sakakura, Miho; Nishioka, Junji; Tomatsu, Hiroaki; Hamaguchi, Yukio; Oguni, Shinichiro; Shiku, Hiroshi

doi:10.1177/107602960501100304

Cited by 19 publications

(14 citation statements)

References 17 publications

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“…If during the course of treatment or monitoring atypical features develop-for example, abnormalities in the white blood cell count, lymphadenopathy, multiple cytopenias-then the diagnosis of ITP should be reassessed. As in childhood ITP, we found insufficient evidence to recommend or suggest the routine use of antiplatelet, [61][62][63][64][65][66][67][68][69][70][71] antiphospholipid [72][73][74][75] and antinuclear antibodies, 13,76 thrombopoietin levels, 64,65,77 or platelet parameters obtained on automated analyzers 64,65,[78][79][80][81][82][83][84] in the evaluation of patients with suspected ITP.…”

Section: Question: What Testing Is Required To Confirm the Diagnosis mentioning

confidence: 99%

The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia

Neunert

Lim

Crowther

et al. 2011

Blood

1,664

1,783

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Immune thrombocytopenia (ITP) is commonly encountered in clinical practice. In 1996 the American Society of Hematology published a landmark guidance paper designed to assist clinicians in the management of this disorder. Since 1996 there have been numerous advances in the management of both adult and pediatric ITP. These changes mandated an update in the guidelines. This guideline uses a rigorous, evidence-based approach to the location, interpretation, and presentation of the available evidence. We have endeavored to identify, abstract, and present all available methodologically rigorous data informing the treatment of ITP. We provide evidence-based treatment recommendations using the GRADE system in those areas in which such evidence exists. We do not provide evidence in those areas in which evidence is lacking, or is of lower quality-interested readers are referred to a number of recent, consensus-based recommendations for expert opinion in these clinical areas. Our review identified the need for additional studies in many key areas of the therapy of ITP such as comparative studies of "front-line" therapy for ITP, the management of serious bleeding in patients with ITP, and studies that will provide guidance about which therapy should be used as salvage therapy for patients after failure of a first-line intervention. (Blood. 2011;117(16):4190-4207)

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Section: Question: What Testing Is Required To Confirm the Diagnosis mentioning

confidence: 99%

The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia

Neunert

Lim

Crowther

et al. 2011

Blood

1,664

1,783

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“…Though automated methods have been available on some Sysmex and Abbott hematology analyzers for some time, there are distinct methodological differences between them, which has further contributed to slow adoption of the assay by clinicians (7)(8)(9)(10). The lack of consensus over the selection of dye (11)(12)(13), the temperature and duration of incubations (13)(14)(15) and the use of whole blood (16) versus platelet rich plasma (14) has further contributed to the slow adoption of this promising measurement. Previous studies determining RP have used various methods for gating this immature fraction of platelets.…”

Section: Introductionmentioning

confidence: 99%

Combined accurate platelet enumeration and reticulated platelet determination by flow cytometry

Hedley

Llewellyn-Smith

Lang

et al. 2015

Cytometry Part B Clinical

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Background: Diagnosing the cause of thrombocytopenia often requires a bone marrow aspiration or biopsy, an invasive procedure. Reticulated platelets (RP) are immature RNA containing platelets, accurate RP enumeration has yet to be achieved, partially due to the lack of a robust reference method. Goal: To refine previous work and gating strategies distinguishing RP from mature platelets while incorporating accurate platelet enumeration into the analysis. After reviewing previously published studies on Thiazole Orange (TO) staining of RP, we systematically evaluated CD41/CD61 in combination with a commercial source of TO (BDBiosciences). Previous RP methods have not taken advantage of platelet enumeration therefore our goal was to incorporate the ICSH platelet enumeration protocol into our method.Methods: TO concentration, incubation, and fixation method were determined to be 10% of stock concentration, 30 min, and 1% formaldehyde respectively. Gating strategy to determine RP fraction used an unstained control tube to set the limit of TO staining.Results: Normal range (n = 51) was 9.9 6 3.1%. Analysis of 40 patients with immunethrombocytopenia-purpura (ITP) showed a RP range from 4.3% to 81.2%. Platelet enumeration was consistent with our previous studies in this area. INTRODUCTIONReticulated platelets (RP), first recognized in 1969 (1), are young, RNA-containing platelets, freshly released from the bone-marrow into the peripheral circulation: RP are analogous to the reticulocytes of erythropoiesis. Subsequent studies showed RP were readily stained with nucleic acid dyes such as Thiazole Orange (TO) (2-4) and that RP have obvious potential as a sensitive early predictor of bone marrow engraftment (5,6). However difficulties with consistency of analysis have hampered wide-spread adoption. The nucleic acid dye based flow methods are problematic to control and these difficulties with specificity and reproducibility have prevented routine adoption of RP counts in the clinical setting. Though automated methods have been available on some Sysmex and Abbott hematology analyzers for some time, there are distinct methodological differences between them, which has further contributed to slow adoption of the assay by clinicians (7-10). The lack of consensus over the selection of dye (11-13), the temperature and duration of incubations (13-15) and the use of whole blood (16) versus platelet rich plasma (14) has further contributed to the slow adoption of this promising measurement. Previous studies determining RP have used various methods for gating this immature fraction of platelets. Work by Ault et al. (3) and Kienast and Schmitz (2) evaluated TO as a stain for RP, as the ability to enter cells without pretreatment and large increase in fluorescent emission upon binding made it ideal. Studies in murine and human samples demonstrated that this was predominately due to RNA content. However, TO fluorescence in platelets is not entirely due to binding of dye to mRNA: Robinson et al. (17) demonstrated a proportion of fluores...

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“…The number of reticulated platelets, which represent the most recently released platelets with a high remaining RNA content, has been introduced as a marker reflecting thrombopoiesis and platelet turnover [19] and may be used as a means of platelet regeneration capacity [20]. Principle and use: Reticulated platelets can be quantified by flow cytometry directly from whole blood without manipulation.…”

Section: Estimation Of Platelet Turnovermentioning

confidence: 99%

Diagnostic Methods for Platelet Function Analysis

Klouche¹

2007

Transfus Med Hemother

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Platelets play a key role in the regulation of haemostasis, clot stability and retraction, vascular repair, and exert an increasingly recognised function in host defence. Conversely, platelets are also implied in many pathophysiological processes, such as thrombosis, haemorrhage, inflammation, atherogenesis, tumour growth and metastasis. Thus quantitative and qualitative platelet alterations have been associated with distinct clinical diseases, including cardiovascular diseases, diabetes, tumour metastasis and sepsis, and consequently different diagnostic platelet markers are correlated with the prognosis and severity of these diseases. While primary platelet functional abnormalities are very rare diseases, acquired platelet function disorders are rather frequent and occur in the context of many diseases and in association with several drugs. Furthermore, the introduction of a great variety of different anti-platelet agents leads to iatrogenic states of therapeutically reduced platelet function. This review provides an overview of the methodological principles of platelet function analysis, the currently available diagnostic devices, the application and suitability for the diagnosis of different platelet function disorders and the restrictions of each method, to promote an optimal selection of diagnostic tests.

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Usefulness of Fully Automated Measurement of Reticulated Platelets Using Whole Blood

Cited by 19 publications

References 17 publications

The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia

The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia

Combined accurate platelet enumeration and reticulated platelet determination by flow cytometry

Diagnostic Methods for Platelet Function Analysis

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