title: Rapid determination of chimerism status using dihydrorhodamine assay in a patient with X-linked chronic granulomatous disease following hematopoietic stem cell transplantation.
abstract: Chronic granulomatous disease (CGD) is a rare genetic disease, which is caused by defects in the NADPH oxidase complex (gp91(phox), p22(phox), p40(phox), p47(phox), and p67(phox)) of phagocytes. This defect results in impaired production of superoxide anions and other reactive oxygen species (ROS), which are necessary for killing bacterial and fungal microorganisms and leads to recurrent, life-threatening bacterial and fungal infections and granulomatous inflammation. The dihydrorhodamine (DHR) flow cytometry assay is a useful diagnostic tool for CGD that can detect absent or reduced NADPH oxidase activity in stimulated phagocytes. We report a patient with X-linked CGD carrying a novel mutation of the CYBB gene whose chimerism status following hematopoietic stem cell transplantation (HSCT) has been rapidly determined using the DHR assay. The level of DHR activity correlates well with short tandem repeat PCR analysis. Considering the advantages of this simple, rapid, and cost-effective procedure, serial measurement of DHR assay would facilitate the rapid determination of a patient's engraftment status, as a supplementary monitoring tool of chimerism status following HSCT.
authors.affiliation: Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
authors.forename: Hyun-Young
authors.initials: HY
authors.lastname: Kim
authors.forename: Hee-Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Chang-Seok
authors.initials: CS
authors.lastname: Ki
authors.forename: Dae Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Keon Hee
authors.initials: KH
authors.lastname: Yoo
authors.forename: Eun-Suk
authors.initials: ES
authors.lastname: Kang
mesh_terms: D001483:Base Sequence; D046528:Chimerism; D004252:DNA Mutational Analysis; D005434:Flow Cytometry; D006105:Granulomatous Disease, Chronic / Q000175:diagnosis* / Q000201:enzymology* / Q000235:genetics / Q000601:surgery; D018380:Hematopoietic Stem Cell Transplantation; D006720:Homozygote; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics*; D009154:Mutation; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000737:chemistry / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012235:Rhodamines / Q000737:chemistry / Q000378:metabolism
chemical_list: D008562:Membrane Glycoproteins; D012235:Rhodamines; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases
keywords: Chimerism; Chronic granulomatous disease; Dihydrorhodamine assay