Assessment of trisomy 12 in chronic lymphocytic leukemia by three methods: conventional fluorescence in-situ hybridization (FISH), FISH in suspension (FISH-IS) and Laser Scanning Cytometry (LSC)

Cuc Do, Karen Lower, Cindy Macardle, Bryone Kuss

    Research output: Contribution to journalMeeting Abstract

    Abstract

    Abstract Details: Chronic lymphocytic leukemia (CLL) is
    the most common adult leukemia in Western countries
    and has an extremely variable clinical course, presumably
    related to its genetic heterogeneity. It is highly
    recommended to conduct genomic analysis before
    initial treatment, including del(13q), trisomy 12,
    del(11q) and del(17p), which are identified by FISH.
    Conventional FISH protocols are labour intensive and
    analyse low total numbers of cells. We compared the
    following 3 technical approaches for specificity, sensitivity
    and cost/time efficiency using FISH as the accepted
    standard:
    (i) Conventional FISH assesses the probes by fluorescence
    microscopy and visually scores up to 200
    nuclei. This technique is considered the diagnostic
    standard assay for CLL despite its limitation of the
    low number of cells targeted and its labour
    intensive demand.
    (ii) FISH in suspension (FISH-IS) incorporates the standard
    FISH technique with the high-throughput
    detection afforded by flow cytometry, enabling
    the quantification of genetic abnormalities in up to 1000 cells/second with high accuracy. Particularly,
    the spot count program in the IDEAS software
    enables accurate scoring of the spots within a cell.
    (iii) Laser scanning cytometry (iCys) is a microscopebased
    cyto-fluorometer that allows quantitative
    measurements on single cells at a high resolution
    with computer controlled analysis of thousands of
    cells. Automated spot counting can solve the
    problem of fatigue caused by many hours of
    microscope viewing, which can lead to mis-interpretation
    and scoring inconsistencies.
    We compared the above methods in the analysis of
    trisomy 12 using the Vysis commercial probe (Abbott
    Molecular) in CLL samples. Centromere 12 probes are
    repetitive sequences that provide very high fluorescence
    intensity at the target site due to a large number of
    bound probes. We were able to confirm that the FISH-IS
    method was able to accurately detect disomy and
    trisomy in CLL cells with centromere 12 probes
    (Figure 1). In addition, iCys analysis is also able to
    confirm the conventional FISH results with nearly 2000
    cells-counted. This research is being continued; the next
    phase is to apply these three methods to locus specific
    probes (13q, 11q, 17p) in order to further evaluate the
    FISH technique before making any conclusions on
    prognosis; and more importantly, its potential use in
    monitoring clonal evolution in CLL.
    Original languageEnglish
    Article numberAbstract 80
    Pages (from-to)61-61
    Number of pages2
    JournalLeukemia and Lymphoma
    Volume56
    Issue numberS1
    DOIs
    Publication statusPublished - 19 Jan 2016
    EventXVI International Workshop on Chronic Lymphocytic Leukemia 2015 - Sydney, Australia
    Duration: 6 Sep 20159 Sep 2015

    Keywords

    • trisomy 12
    • lymphocytic leukemia
    • FISH-IS

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