title: A multicenter investigation with interphase fluorescence in situ hybridization using X- and Y-chromosome probes.
abstract: Twenty-six laboratories used X and Y chromosome probes and the same procedures to process and examine 15,600 metaphases and 49,400 interphases from Phaseolus vulgaris-leucoagglutinin (PHA)-stimulated lymphocytes. In Part I, each laboratory scored 50 metaphases and 200 interphases from a normal male and a normal female from its own practice. In Part II, each laboratory scored 50 metaphases and 200 interphases on slides prepared by a central laboratory from a normal male and a normal female and three mixtures of cells from the male and female. In Part III, each laboratory scored 50 metaphases (in samples of 5, 10, 15, and 20) and 100 interphases (in samples of 5, 10, 15, 20, and 50) on new, coded slides of the same specimens used in Part II. Metaphases from male specimens were scored as 98-99% XY with no XX cells, and 97-98% of interphases were scored as XY with 0.04% XX cells. Metaphases from female specimens were scored as 96-97% XX with 0.03% XY cells, and 94-96% of interphases were scored as XX with 0.05% XY cells. Considering the data as a model for any probe used with fluorescence in situ hybridization (FISH), a statistical approach assessing the impact of analytical sensitivity on the numbers of observations required to assay for potential mosaicisms and chimerisms is discussed. The workload associated with processing slides and scoring 50 metaphases and 200 interphases using FISH averaged 27.1 and 28.6 minutes, respectively. This study indicates that multiple laboratories can test/develop guidelines for the rapid, efficacious, and cost-effective integration of FISH into clinical service.
authors.affiliation: Mayo Clinic, Rochester, Minnesota 55905, USA.
authors.forename: G
authors.initials: G
authors.lastname: Dewald
authors.forename: R
authors.initials: R
authors.lastname: Stallard
authors.forename: A
authors.initials: A
authors.lastname: Al Saadi
authors.forename: S
authors.initials: S
authors.lastname: Arnold
authors.forename: P I
authors.initials: PI
authors.lastname: Bader
authors.forename: R
authors.initials: R
authors.lastname: Blough
authors.forename: K
authors.initials: K
authors.lastname: Chen
authors.forename: B R
authors.initials: BR
authors.lastname: Elejalde
authors.forename: C J
authors.initials: CJ
authors.lastname: Harris
authors.forename: R R
authors.initials: RR
authors.lastname: Higgins
authors.forename: G A
authors.initials: GA
authors.lastname: Hoeltge
authors.forename: W T
authors.initials: WT
authors.lastname: Hsu
authors.forename: V
authors.initials: V
authors.lastname: Kubic
authors.forename: D J
authors.initials: DJ
authors.lastname: McCorquodale
authors.forename: M A
authors.initials: MA
authors.lastname: Micale
authors.forename: J W
authors.initials: JW
authors.lastname: Moore
authors.forename: R M
authors.initials: RM
authors.lastname: Phillips
authors.forename: S
authors.initials: S
authors.lastname: Scheib-Wixted
authors.forename: S
authors.initials: S
authors.lastname: Schwartz
authors.forename: S
authors.initials: S
authors.lastname: Siembieda
authors.forename: K
authors.initials: K
authors.lastname: Strole
authors.forename: P
authors.initials: P
authors.lastname: VanTuinen
authors.forename: G H
authors.initials: GH
authors.lastname: Vance
authors.forename: A
authors.initials: A
authors.lastname: Wiktor
authors.forename: A
authors.initials: A
authors.lastname: Zinsmeister
mesh_terms: D003582:Cytogenetics / Q000592:standards; D015342:DNA Probes; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation / Q000379:methods*; D007399:Interphase; D007753:Laboratories / Q000592:standards; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology; D008297:Male; D008677:Metaphase; D010835:Phytohemagglutinins; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016526:Workload; D014960:X Chromosome; D014998:Y Chromosome
chemical_list: D015342:DNA Probes; D010835:Phytohemagglutinins