A simple flow cytometry assay using dihydrorhodamine for the measurement of the neutrophil respiratory burst in whole blood: comparison with the quantitative nitrobluetetrazolium test.
The neutrophil respiratory burst is essential for the host's ability to kill ingested microorganisms. Several flow cytometric assays have recently been developed to measure this process. These assays are largely unvalidated. In this study a whole blood flow cytometry assay using dihydrorhodamine 123 (DHR) as a substrate was compared with the quantitative nitrobluetetrazolium (NBT) test, an accepted measure of the earliest events in the respiratory burst. Because whole blood is used, the new assay is quicker and simpler than existing flow cytometry assays. Specimens as small as 0.1 ml can be used which makes the assay ideal for use in neonates and young children. There was a high degree of correlation between the DHR assay and the quantitative NBT test (r(s) = 0.76, P < 0.01). It is concluded that the whole blood DHR assay is an accurate and sensitive measure of the respiratory burst.
Richardson
M P
MP
Paediatric Infectious Diseases Unit, St. George's Hospital, London, UK. mrichard@sghms.ac.uk
Ayliffe
M J
MJ
Helbert
M
M
Davies
E G
EG
D000328:Adult; D001769:Blood; D005434:Flow Cytometry / Q000379:methods*; D005562:Formazans / Q000032:analysis; D006801:Humans; D018375:Neutrophil Activation; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D009580:Nitroblue Tetrazolium / Q000378:metabolism*; D016897:Respiratory Burst; D012235:Rhodamines / Q000032:analysis / Q000378:metabolism*; D013053:Spectrophotometry; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors
D005562:Formazans; D012235:Rhodamines; C058319:dihydrorhodamine 123; D009580:Nitroblue Tetrazolium; D013755:Tetradecanoylphorbol Acetate