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Clinical and Diagnostic Laboratory Immunology, 03 1995, 227-232, Vol 2, No. 2
Copyright © 1995 by the American Society for Microbiology. All rights reserved.

Rapid whole-blood flow cytometry assay for diagnosis of chronic granulomatous disease

MR O'Gorman and V Corrochano
Department of Pediatrics, Northwestern University, Children's Memorial Hospital, Chicago, Illinois 60614.

Chronic granulomatous disease (CGD) is characterized by defective killing of intracellular microorganisms due to mutations in one of the four known components of the NADPH oxidase system. This system is responsible for the generation of superoxide and related antimicrobial oxidants. Diagnosis of CGD requires the demonstration of an abnormal oxidase system in the leukocytes of affected patients. Recently, several flow cytometry-based procedures which measure various reactive oxygen intermediates generated by the NADPH oxidase system have been developed. Most of the procedures developed to date require time- consuming granulocyte isolation, washing, and counting procedures, or they lack sensitivity. We have modified an existing procedure such that cell labelling and stimulation are performed directly in whole blood. Optimization of this procedure and its use in the diagnosis of patients with CGD or X-linked carriers are presented.

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