Method for determining the haemoglobin F content of an erythroid cell
AssigneesUniversite Paris Est Creteil Val De Marne, Institut National De La Sante Et De Recherche Medicale (Inserm), Etablissement Francais Du Sang, Assistance Publique-Hopitaux de Paris
InventorsMarie Cambot, Gaetana Vandemeulebrouck, France Noizat Pirenne, Pablo Bartolucci, Marie Georgine Rakotoson, Frédéric Galacteros, Nicolas Hebert
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Abstract
The invention concerns a method for determining, by flow cytometry, the hemoglobin F (HbF) content of each erythroid cell of a set of erythroid cells. This method applies in particular to determining the HbF content of each red blood cell of a set of red blood cells. The invention also concerns a method for determining the amount of red blood cells transfused into a patient and for monitoring the therapeutic efficacy of a treatment for sickle cell disease or β-thalassemia.
Claims16
A method for determining, in vitro, a hemoglobin F (HbF) content of each erythroid cell of a set of erythroid cells contained in a sample of erythroid cells, comprising the steps of:
- a) isolating erythroid cells from the sample;
- b) permeabilizing the membrane of the isolated erythroid cells;
- c) labeling the HbF of the erythroid cells obtained in step b) with an anti-HbF antibody conjugated to a fluorochrome capable of emitting fluorescence;
- d) measuring, by flow cytometry, fluorescence intensity (MFI) of each erythroid cell of the set of erythroid cells;
- and
- e) determining the hemoglobin F content of each erythroid cell of the set of erythroid cells by comparing the fluorescence intensity of each erythroid cell with a standard curve associating the fluorescence intensity measured for an erythroid cell with the HbF content.
Dependencies are inferred from the claim text.