Practical Guidelines and Tips for Sensitive and Accurate Identification of PHN Clones

Since flow cytometry has the ability to rapidly detect and quantify GPI-deficient cells, this method has become the "gold standard" for the detection of PNH clones, which has led to improved diagnosis, patient management, and prognosis in PNH and related disorders. Traditionally this was performed using the antibodies CD55 and CD59 as they are able to detect GPI-deficient cells in both the RBCs and WBC. However, the sensitivity this approach was poor and the false-positive as well as the false-negative rate rendered this method inadequate for reliable detection of PNH clones. In the more recent past, more sensitive approaches have evolved due to the utilization of better-suited GPI-linked antibodies and new reagents such as FLAER (bacterial lysin). Accurate detection, monitoring, and diagnosis of PNH have become increasingly important priorities for clinical flow laboratories performing PNH screening. This presentation will discuss the important steps leading to a sensitive and accurate PNH assay: Optimisation of instruments, targeted selection of antibody clones, set-up & analysis and reporting based on the 2012 ICCS Guidelines as well as the 2012 Practical PNH Guidelines

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