title: Long polymerase chain reaction-based fluorescence in situ hybridization analysis of female carriers of X-linked chronic granulomatous disease deletions.
abstract: Chronic granulomatous disease (CGD) is a rare inherited disorder in which antimicrobial activity of phagocytes is impaired due to the lack of reactive oxygen species, or oxidative burst, produced by NADPH oxidase. The X-linked form of CGD, representing approximately 70% of all cases, is caused by mutations in the cytochrome b beta subunit (CYBB) gene, which maps to chromosome Xp21.1. CYBB encodes the gp91-phox protein, a necessary component in the NADPH oxidase pathway. A wide variety of mutations have been identified in X-linked CGD patients, all of which lead to deletion of the functional protein and no oxidative burst activity. The mutations vary from single nucleotide substitutions to deletions of the entire gene. In this article, we report a mutation detection method for probands of female relatives at risk for carrier status of large deletions of the CYBB gene. Through fluorescent in situ hybridization of metaphase chromosomes, we were able to consistently distinguish carriers from noncarriers using polymerase chain reaction-derived, labeled DNA specific for exons 2 to 13 of the CYBB region at Xp21.1.
authors.affiliation: Lombardi Cancer Center, Georgetown University Medical Center, 3800 Reservoir Rd., NW, IMHG/M4000, Washington, Distric of Columbia 20007, USA.
authors.forename: Kelly Claire
authors.initials: KC
authors.lastname: Simon
authors.forename: Deborah
authors.initials: D
authors.lastname: Noack
authors.forename: Julie
authors.initials: J
authors.lastname: Rae
authors.forename: John
authors.initials: J
authors.lastname: Curnutte
authors.forename: Shireen
authors.initials: S
authors.lastname: Sarraf
authors.forename: Valentin
authors.initials: V
authors.lastname: Kolev
authors.forename: Jan K
authors.initials: JK
authors.lastname: Blancato
mesh_terms: D005091:Exons; D005260:Female; D017353:Gene Deletion; D006580:Genetic Carrier Screening / Q000379:methods*; D006105:Granulomatous Disease, Chronic / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics*; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D008562:Membrane Glycoproteins; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases