The Role of Flow Cytometry in the Diagnosis of Paroxysmal Nocturnal Hemoglobinuria in the Clinical Laboratory
Section snippets
Flow cytometry: general considerations
The availability of a diagnostic flow cytometry assay means that for patients who have suspected PNH, a definitive diagnosis can be established rapidly, resulting in improved patient management and prognosis. As PNH is a rare condition, it is essential to demonstrate deficiency of at least two GPI-linked antigens from red cells and granulocytes to establish a diagnosis confidently. For granulocytes, this is achieved best by multicolor cytometry, although more complex combinations allow
Diagnosis of paroxysmal nocturnal hemoglobinuria by flow cytometry: clinical value
Once a diagnosis of PNH is established on the basis of clinical findings and flow cytometry results, serial monitoring of the peripheral blood PNH clone can provide important information that may influence patient management [28]. Patients who have type III red cells (greater than 20%) almost always show signs of intravascular hemolysis, whereas patients who have granulocyte PNH clones (>50%) have a significant risk of thrombosis, and prophylactic anticoagulation is an important consideration
Internal and external quality assurance
As with any diagnostic flow cytometry assay, internal and external quality assurance forms an integral part of good clinical laboratory practice. Essential steps in establishing the validity of the assay should include extensive characterization of the antibody test combinations on normal peripheral blood samples and then validated on patients who have a confirmed diagnosis of PNH. As screening for PNH by flow cytometry is a clinical diagnostic test, but may be requested infrequently, it is
Summary
PNH is a rare disease in which detection of GPI-deficient blood cells (PNH clones) by flow cytometry is an essential component of the diagnostic process. Recently, the role of flow cytometry in this disorder has been extended to play a pivotal role in the clinical trial and assessment of treatment response of a novel monoclonal antibody therapy for PNH [36]. Eculizumab is a monoclonal antibody against the complement component C5 and has been used successfully to inhibit intravascular hemolysis
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