Dear Sebastian, I will first briefly explain the situation within our institute to clarify why we perform the PNH test as described. CLB (Central Laboratory of the Dutch Red Cross Blood transfusion services), situated in Amsterdam, is with more than 800 co-workers active in the following fields: 1. Preparation of products: therapeutic products derived from blood and production of blood grouping and immune reagents (CD's, cytokines, a.o.). 2. Specialised laboratory services for Dutch hospitals, including: . blood typing; . transfusion reactions due to the administration of blood and blood products; . identification of abnormalities in blood cells and plasma proteins. 3. Applied and fundamental research in the following areas: . homeostasis and thrombosis; . transfusion medicine and haematology; . inflammation and sepsis; . clinical and experimental immunology; . product development. 4. Academic and postgraduate teaching of students in medicine, biochemistry and biology. 5. Reference tasks include: "WHO International Laboratory for Biological Standards with special reference to human blood products and blood derivatives" and the "European Bank of Frozen Blood of Rare Groups". You can imagine that this combination of fields for PNH has given us the opportunity to develop a very accurate flow cytometry assay. This is published by Ellen van de Schoot at al. in Blood, 1990, 1853-1859 : Deficiency of PI-linked membrane glycoproteins of leukocytes in PNH, Description of a new diagnostic cytofluorometric assay. In this study it is shown that the expression of PI markers is most reliable measured at granulocytes and there fore we use at present a panel of two colour antibody combinations. We use CD66e-PE as granulocyte-identification marker and CD16, CD24, CD66b and CD59 as PI markers next to a negative control and two leukaemia related controls: CD15 and CD11b (all FITC). Of course all antibodies are in-house made, well characterised in the international workshops and commercially available. Some times we had a remark that so many antibodies makes the PNH test very expensive, but if you realise that all antibodies can be diluted 1:20 and thus you can perform 2000 test with one vial which makes the cost almost negligible. For information on our products and distributors you can check our web site, for clinical information you can contact Dr. Ellen van der Schoot directly at CLB tel +31-20-5123377. Or you can (as many hospitals do) send your patient sample to CLB-diagnostic services. John Voorn CLB Reagents Productmanager Plesmanlaan 125 j_voorn@clb.nl <mailto:j_voorn@clb.nl> 1066CX AMSTERDAM the Netherlands tel +31-(0)20 5123246 fax +31-(0)20 5123570 web site www.clb.nl <www.clb.nl>
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