title: Interphase Chromosome Flow-FISH.
abstract: A 2-day method using flow cytometry and FISH for interphase cells was developed to detect monosomy 7 cells in myelodysplastic syndrome patients. The method, Interphase Chromosome Flow-FISH (IC Flow-FISH), involves fixation of leukocytes from blood, membrane permeabilization, hybridization of cellular DNA with peptide nucleic acid probes with cells intact, and analysis by flow cytometry. Hundreds to thousands of monosomy 7 cells were consistently detected from 10-20 mL of blood in patients with monosomy 7. Proportions of monosomy 7 cells detected in IC Flow-FISH were compared with results from conventional cytogenetics; identification of monosomy 7 populations was verified with FACS; and patient and donor cells were mixed to test for sensitivity. IC Flow-FISH allows for detecting monosomy 7 without requiring bone marrow procurement or the necessity of metaphase spreads, and wider applications to other chromosomal abnormalities are in development.
authors.affiliation: Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Keyvan
authors.initials: K
authors.lastname: Keyvanfar
authors.forename: Jason
authors.initials: J
authors.lastname: Weed
authors.forename: Prashanth
authors.initials: P
authors.lastname: Swamy
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kajigaya
authors.forename: Rodrigo T
authors.initials: RT
authors.lastname: Calado
authors.forename: Neal S
authors.initials: NS
authors.lastname: Young
mesh_terms: D000328:Adult; D000368:Aged; D016022:Case-Control Studies; D002869:Chromosome Aberrations; D002872:Chromosome Deletion; D002897:Chromosomes, Human, Pair 7 / Q000235:genetics; D015342:DNA Probes; D005434:Flow Cytometry; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics*; D007621:Karyotyping; D008677:Metaphase / Q000235:genetics*; D008875:Middle Aged; D009000:Monocytes / Q000473:pathology; D009190:Myelodysplastic Syndromes / Q000175:diagnosis* / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D055815:Young Adult
chemical_list: D015342:DNA Probes