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Use of Flow Cytometry Immunophenotyping For Diagnosis of Leukemia at Moi Teaching and Referral Hospital
Use of Flow Cytometry Immunophenotyping For Diagnosis of Leukemia at Moi Teaching and Referral Hospital
Back ground
World Health Organization classification of
Backg.
REAL classification of lymphoid was based
Introduction
Flow cytometry is a technology that
Specimen processing
Specimens commonly analyzed are bone
Spec
A cell count is obtained before flow is run-
Analysis
A measurement for the diffraction of
Data
Acute promyelocytic
leukemia
Study
Objective: The main objective of the study was to
Morphological diagnosis
The original classification scheme proposed
Flow cytometry
Equipment: Four colour computerized BD FACS.
The fluorochromes FITC, PE, Per CP, and APC
were applied
Panel of monoclonal antibodies used include:
-Pan Leukocyte antigen: CD 45 is a pan leukocyte
marker
- B- cell CD10, CD19, CD20, cyt CD79a
-T cell; CD3, CD4, CD7, CD8, cyt CD3
-Myeloid; CD33, CD117, MPO.
-Immature cell antigens; CD34, and HLADR
-Erythroid marker: CD71
Results
The total number of samples run between
Results
Morphology
Flow +ve
-ve
+ve
-ve
Total
12
18
Total 15
11
14
17
32
Results
Morphology
+ve
-ve
Total
10
13
23
Total 17
15
32
Flow +ve
-ve
Morpholog Flow
y
Comment
003
AML M5
CD45(dim)-95%,
CD33(95%), CD38(86%),
HLADR(92%),
CytCD3(95%), MPO-4%
-Reported as T-ALL
Biphenotypic
006
AML M5
CD 45(dim)-21%,
cytCD7a-21%-Reported
as B-ALL
Few events70,000
007
Inconclusive CD45(dim)-66%,
CD38(51%),CD33(50%)
HLADR(43%)-Reported
as AML
Inadequate for
morphological
evaluation
016
AML M2
CD45(bright)-30%,
CD5(31%)
Few events15,000
018
ALL
CD45dim)-73%,
CD5(61%), cyt
Biphenotypic
Morpholog Flow
y
Comment
019
AML M1
CD45(dim)-82%,
CD33(74%),
HLADR(73%),
cytCD3(93%)-Reported
as T-ALL
Biphenotypic
024
ALL
CD45 (bright)-28%,
CD3(20%)-Inconclusive
results
Few events10,000
025
AML M1
CD45(bright)54%,CD45(dim)-22%
CD19(24%), CD3(54%),
CD5(52%),HLADR-35%,
CD79a(23%)-Reported
as T-ALL
Few events10,000
WHO-2008:
MWWHOHixeWenotype Acute
LeukemiaWHOW
B cell lineage
T cell lineage
Myeloid lineage
Strong CD19
with :
cCD22, CD10 or
cCD79a
CD3 (c or s)
cMPO
2 or more of:
CD11c, CD14,
CD36, CD64
Flow cytometry
Few events<100,000
Poor sample collection
Abberant expression of some
markers
Few flow cytometrists
Consultation services
Conclusions
Flow cytometry is an important diagnostic
Acknowledgments
Moi University /MTRH-Kirtika Patel (PI),