Enumeration of Low White Cells

  • Girolamo Sirchia
  • Paolo Rebulla
Chapter
Part of the Medical Intelligence Unit book series (MIUN)

Summary

White blood cells (WBC) present in blood and blood components can cause a number of posttransfusion complications. At present, filtration is the most popular method developed to prepare blood components depleted of WBC. Current filters are capable of removing more than 99.9% WBC (3 logl0 removal). Consequently, filtered products can contain as few as 1–10 WBC/µL (300,000–3,000,000 in a 300 mL unit), or less. At these levels routine WBC counting methods are greatly inaccurate and imprecise, as supported by theoretical considerations already presented in the medical literature almost 50 years ago. These considerations indicate that: (1) the error of counting approximates the square root of the number of cells counted. As a consequence, counting methods should aim at examining large sample volumes, so as to collect the greatest possible number of cells; (2) rare events, such as WBC randomly distributed in a counting chamber, follow the Poisson distribution. This distribution can be used to determine the level of confidence one can have that a certain count is below a certain level (upper confidence limit of that count). In reporting the results of evaluations performed in leukodepleted blood components this level should be preferred to mean or median counts, since at the clinical level it is important that the recipient receives less than rather than an average number of WBC.

Methods specifically designed for counting residual WBC include: (1) microscopic methods performed with traditional or large volume counting chambers; (2) flow cytometry procedures which use light scatter and/or DNA staining techniques; (3) radioimmunoassays; (4) techniques based on leukocyte DNA amplification and (5) a qualitative method based on trapping residual leukocytes in 3 µm pore size polycarbonate filters. Of these, counting in large volume chambers such as the Nageotte chamber has been recommended as the method offering the best compromise of precision and feasibility for routine quality control. Flow cytometry, which requires access to expensive equipment, shows similar levels of precision and accuracy and slightly better sensitivity. In view of the continuous improvement of filters for leukodepletion, better counting methods should be developed. In addition, since the prevention of different complications seems to require the achievement of different levels of leukodepletion, specific protocols for quality assurance should be designed in relation to the type of complication to be prevented (e.g. non-hemolytic, febrile transfusion reaction, CMV transmission, etc).

Finally, efforts to develop improved methods for counting residual WBC in leukodepleted blood components should be accompanied by conclusive studies designed to increase our knowledge on the minimum number of residual WBC capable of inducing different posttransfusion complications.

Keywords

White Blood Cell Blood Component Limit Dilution Assay Current Filter Pore Size Polycarbonate Filter 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer-Verlag Berlin Heidelberg 1995

Authors and Affiliations

  • Girolamo Sirchia
  • Paolo Rebulla

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