Chapter 11.18 : Flow Cytometric Test for Chronic Granulomatous Disease

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Normal neutrophil function is of great importance in the host defense against bacterial and fungal infections. As neutrophils recognize, adhere to, and phagocytose an invading organism, they generate an oxidative burst which results in the reduction of molecular oxygen to superoxide. The superoxide produced is rapidly converted to hydrogen peroxide (HO, which kills the pathogen). In chronic granulomatous disease (CGD), microbial killing is defective because neutrophils from patients with CGD lack a respiratory burst. The assay described here uses dihydrorhodamine-123 (DHR-123) and phorbol 12 myristate 13 acetate (PMA) to measure oxidative burst activity, which is easily converted to hydrogen peroxide. DHR, a nonfluorescent compound, reacts with hydrogen peroxide and is oxidized to rhodamine-123, a green fluorescent compound. Cellular fluorescence is measured using flow cytometry.

Citation: Garcia L. 2010. Flow Cytometric Test for Chronic Granulomatous Disease, p 254-257. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.18
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1. Bass, D.,, W. Parce,, L. Dechatelet,, P. Szejda,, M. Seeds,, and M. Thomas. 1983. Flow cytometric studies of oxidative product formation by neutrophils: a grade response to membrane stimulation. J. Immunol. 136: 1910 1917.
2. Centers for Disease Control and Prevention. 1997. Revised guidelines for performing CD4 + T-cell determinations in persons infected with human immunodeficiency virus (HIV). MMWR Morb. Mortal. Wkly. Rep. 46( RR-2): 129.
3. Emmendorffer, A.,, M. Hecht,, M. L. Lohmann-Matthes,, and J. Roesler. 1990. A fast and easy method to determine the production of reactive oxygen intermediates by human phagocytes using dihydrorhodamine 123. J. Immunol. Methods 131: 269 275.
4. Hassan, N. F.,, D. E. Campbell,, and S. D. Douglas. 1988. Phorbol myristate acetate oxidation of 2′7′ dichlorofluorescein by neutrophils from patients with chronic granulatomatous disease. J. Leukoc. Biol. 43: 317 322.
5. Malech, H. L.,, and J. I. Gallin. 1987. Neutrophils in human diseases. N. Eng. J. Med. 317: 687 693.
6. O’Gorman, M. R. G.,, and V. Corrochano. 1995. Rapid whole blood flow cytometry assay for the diagnosis of chronic granulomatous disease. Clin. Diagn. Lab. Immunol. 2: 227 232.
7. Vowells, S. J.,, S. Sekhsaria,, H. L. Malech,, M. Shalit,, and T. A. Fleisher. 1995. Flow cytometric analysis of granulocyte respiratory burst: a comparison study of fluorescent probes. J. Immunol. Methods 178: 89 97.

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