Rapid Micromethod for the Preparation of Leucocyte-Free Haemolysates for the Determination of Pyruvate Kinase and other Erythrocyte Enzymes

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Abstract
In pyruvate kinase deficiency the high activity of the leucocyte isoenzyme may mask the erythrocyte defect. Separation of leucocytes from erythrocytes by the commonly used sedimentation or filtration procedures requires rather large volumes of blood, is time-consuming, and sometimes leaves an unsatisfactorily large proportion of leucocytes. In the present study at least 95% of leucocytes, measured with the lysosomal enzyme β-N-acetylhexosaminidase as a very sensitive marker for leucocyte lysis, were consistently removed by differential lysis of erythrocytes in hypotonic saline. Haemolysates suitable for erythrocyte enzyme assays can be prepared from very small quantities of capillary blood (10–100 μl) very rapidly, cheaply, and reproducibly by this method.