title: Standardized flow cytometric method for the accurate determination of platelet counts in patients with severe thrombocytopenia.
abstract: BACKGROUND
  The therapeutic option of prophylactic platelet (PLT) transfusion in cases of severe thrombocytopenia critically depends on the availability of accurate and precise counts because clinical decisions are widely based on decision or trigger points. Although often applied in current practice at a level of 20 Gpt/L, there is increasing evidence that the trigger points could safely be reduced to 10 or even 5 Gpt/L. In order to facilitate this downward revision, it is necessary to have PLT counting methods that are able to provide reliable results in the appropriate decision range.
  
  
  METHODS
  Postchemotherapy-induced pancytopenia PLT counting was performed in patients with hematological malignant disorders. This study describes a novel flow cytometric method that utilizes a PLT-specific monoclonal antibody (CD41a) in conjunction with fluorescent reference beads in order to derive absolute platelet numbers.
  
  
  RESULTS
  Applying a mathematical model, this flow cytometric method was shown to have a detection limit of 0.24 Gpt/L and a lower limit of quantification (coefficient of variation [CV] = 10%) of 1.1 Gpt/L. These values are a substantial improvement on previously reported results for the Technicon H1 automated instrument or manual hemocytometry. Moreover, although the flow cytometry and Technicon H3 methods were found by supplementary analyses to show a reasonably good correlation, the hematology instrument showed a distinct tendency to overestimate PLT counts at low levels.
  
  
  CONCLUSIONS
  It is proposed that this standardized immunoplatelet method offers the best approach in evaluating, at the clinical level, the possibility of lower PLT transfusion triggers. It can be used to evaluate the performance limitations of automated hematology analyzers that are widely used at the present time.
authors.affiliation: Institut für Klinische Chemie und Pathobiochemie, Universitätsklinikum der RWTH, Aachen, Germany. Dagmar.kunz@post.rwth-aachen.de
authors.forename: D
authors.initials: D
authors.lastname: Kunz
authors.forename: H
authors.initials: H
authors.lastname: Höffkes
authors.forename: W S
authors.initials: WS
authors.lastname: Kunz
authors.forename: A M
authors.initials: AM
authors.lastname: Gressner
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D005260:Female; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards* / Q000706:statistics & numerical data; D006801:Humans; D008297:Male; D008875:Middle Aged; D010976:Platelet Count / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D012044:Regression Analysis; D013921:Thrombocytopenia / Q000097:blood*