A simple chemiluminescence assay for the determination of reactive oxygen species produced by human neutrophils

We show that phagocyte production of reactive oxygen species can be measured using a microtitre plate based chemiluminescence blotting technique, The production of reactive oxygen species is determined by their ability to catalyze the oxidation of luminol or isoluminol, resulting in light emission which is recorded on a photographic film. The method permits the determination of NADPH oxidase activity from as few as 9000 cells. It could be used to detect NADPH oxidase defects in neutrophils (e.g. from patients suffering from chronic granulomatous disease), and to screen pharmaceuticals with scavenging activity for reactive oxygen species.