ICSH Expert Panel on Cytometry: International Council for Standardization in Haematology (ICSH) Recommendations for “Surrogate Reference” Method for the Packed Cell Volume

School of Medicine, Department of Pathology and Human Anatomy, Loma Linda University, Loma Linda, CA 92354, USA.
Laboratory Hematology 02/2003; 9(1):1-9.
Source: PubMed


The spun packed cell volume (PCV, hematocrit) is a key measurement on which are based hematology instrument calibration, reference range determination, and assignment of values to calibrators/controls. In 2001, the International Council for Standardization in Haematology (ICSH) recommended a Reference PCV method, which is fully traceable to the ICSH reference hemoglobin method. Because of its complexity, however, this method is impractical for occasional use in routine laboratories and is therefore intended primarily for use by manufacturers of capillary microhematocrit tubes, liquid calibrators, and multichannel analyzers. In response to the need for a simpler method--accessible to all routine laboratories--the ICSH offers this "Surrogate Reference" PCV procedure. It is traceable to the original ICSH Reference PCV method and is based on spun PCVs obtained using borosilicate capillary tubes with an already-known relationship to this reference procedure. This ICSH "Surrogate Reference" PCV method is substantially simpler, thus putting it within the reach of most routine hematology laboratories.

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    • "The normal PCV percentage for goats is ~30%; values ⩽20% indicates anaemia. The PCV was measured using the capillary microhaematocrit method by centrifugation for 5 min at 15 300 × g in capillary tubes (Bull et al., 2003). "
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    • "The remaining portion of the blood sample was submitted to further centrifugation (1000 g, 20 min) to obtain PPP and packed RBC. The packed RBC concentration was measured using a microhaematocrit determination (Bull et al, 2003). Washed platelet concentrates were prepared from PRP by centrifugation (20 min, 1000 g) in the presence of ethylenediaminetetraacetic acid (EDTA) (13 mmol/l final concentration). "
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    ABSTRACT: Red blood cells (RBC) contribute significantly to haemostasis and thrombosis under oscillatory flow conditions, and erythrocytosis has been associated with increased thrombotic risk. To measure the dynamic influences of RBC on platelets, we used a recently described cone and plate(let) analyzer (CPA), evaluating the effect of haematocrit (Hct) on platelet function in whole blood under arterial flow conditions (1800/s, 2 min, 25 degrees C). Anticoagulated blood, reconstituted to varying haematocrits with autologous RBC, demonstrated a significant increase in adherent platelet aggregate formation at Hct levels >45%. This increase was not affected by pretreatment of blood with 0.05 mmol/l aspirin, but was prevented by antagonists of P2Y1, P2Y12, or P2X1, ADP and ATP receptors, and by converting exogenous ADP to ATP with creatine phosphate/creatine phosphokinase. As negligible platelet granule secretion was measured during CPA analysis, but metabolic inhibition of RBC with sodium azide or glutaraldehyde fixation inhibited erythrocytosis-enhanced increases in platelet aggregate size, adenine nucleotides contributing to shear-induced platelet aggregate formation appear to be derived from erythrocytes. These findings support the use of CPA for ex vivo evaluation of the contribution of RBC to platelet function and its inhibition under physiological shear conditions.
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