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october 7-9, 2012 • NEW ORLEANS, LA
using PBS+0.5% (w/v) bovine serum albumin (BSA)
before being resuspended in 200μl PBS+0.5%BSA. One
hundred thousand erythrocyte events were acquired on a
FACS Canto II. When data obtained by flow cytometry was
correlated with visual microscopy looking for spherocytes,
a correlation was evident with rare visual spherocytes
(n=26) showing a mean fluorescence of 9100, occasional
spherocytes (n=10) with a mean of 8700, and moderate
spherocytosis (n=6) with 6800. Samples also appear to be
reasonably stable with the EMA fluorescence decreasing
by approximately 8% 4 days post draw. This assay will be a
useful diagnostic tool for the diagnosis of HS.This work was
funded by the ARUP Institute for Clinical and Experimental
Pathology.
P24
DETECTION OF MINIMAL RESIDUAL DISEASE (MRD)
IN B LYMPHOBLASTIC LEUKEMIA/LYMPHOMA (B-ALL)
USING A 9 PARAMETER KALUZA RADAR PLOT
Joanne M Luider
1
, Karan Paisooksantivatana
1
, Maria C
Gentile2, Iwona A Auer
1
, Adnan Mansoor
1
1
Calgary Laboratory Services,
2
Beckman Coulter Canada
Introduction:
Identification of B-ALL MRD to a sensitivity
level of 0.01% is now routinely required for risk-stratified
treatment protocol as well as for clinical trials. In addition
to all technical issues, data analysis is the most crucial
step for accurate quantification of MRD. For this purpose,
sophisticated and extensive sequential gating strategies are
typically utilized in many laboratories using different analysis
software.
Aim:
To determine if the Kaluza radar-plot (showing
9 parameters in one plot) can simplify MRD analysis for
B-ALL.
Methods:
Ten normal/regenerating bone marrows
and fifty bone marrow samples from both adult (n=27)
and paediatric (n=20) patients were stained in a single
10-colour tube with the following antibodies: CD58-FITC,
CD49f-PE, CD34-ECD, CD38-PC5.5, CD81-APC, CD22-
APC-AF700, CD10-APC-AF750, CD123-PB, and CD45
Kro. All CD19-positive events were gated and subsequently
analyzed for MRD using both a standard sequential gating
strategy and a single 9 parameter Kaluza radar plot. MRD
was identified as a cell cluster (at least 10 events) falling
outside the normal maturation “zone” defined by the 10
reference BM’s.
Results:
All fifty samples, 34 MRD-negative
and 16 MRD-positive bone marrows, showed complete
agreement between the sequential gating strategy and
the single 9-parameter Kaluza radar plot in terms of either
qualitative or quantitative results.
Conclusions:
B-ALL MRD
can be accurately and efficiently quantified using a simple
9-parameter Kaluza radar plot from a single 10-color tube
analysis.
P25
COMBINED TCR GAMMA/DELTA T CELL SUBSETS IN
HEPATOSPLENIC T-CELL LYMPHOMA
Neil V McNamara, Aysha Othman, Anne-Marie Watson,
Lindsay Dunlop, Silvia Ling
Haematology, Liverpool Hospital
Hepatosplenic T-cell lymphoma (HSTL) is a rare and
aggressive disease of cytotoxic T cells expressing T cell
receptor (TCR) gamma/delta. Presentation may include,
fever, abdominal pain and hepatosplenomegally without
lymphadenopathy. Bone marrow involvement is common,
with thrombocytopenia and occasional pancytopenia.
Immunophenotyping by flow cytometry is a principle
diagnostic tool for HSTL. We describe 2 cases of this
rare condition, with immunophenotypes in bone marrow
samples of Case (1): CD2+, CD3+, CD4-, CD5-, CD7+/-
CD8-, CD16+, CD56+, TCR gammadelta+, and Case (2):
POSTER ABSTRACTS