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Diagnosis of Acute Leukemia (AL) by Flow Cytometry (FCM)

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Diagnosis of Acute Leukemia (AL) by Flow Cytometry (FCM) S. Oukid, S. Taoussi, M.T. Abad. Department of Hematology, EHS CAC Blida, Algeria – PowerPoint PPT presentation

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Title: Diagnosis of Acute Leukemia (AL) by Flow Cytometry (FCM)


1
  • Diagnosis of Acute Leukemia (AL) by Flow
    Cytometry (FCM)
  • S.
    Oukid, S. Taoussi, M.T. Abad.
  • Department of
    Hematology, EHS CAC Blida, Algeria
  • ESH International
    conference on AN UPDATE ON THE MANAGEMENT
  • OF HAEMATOLOGICAL
    MALIGNANCIES
  • Hammamet, Tunisia
    October 28 30- 2010

2
  • Objectives
  • A major application of flow cytometry in
    hematology is to
  • characterize the malignant cells in
    acute leukemia for diagnosis,
  • therapeutic decision and assessment of
    prognosis.

3
  • Introduction
  •                 
  •  Flow cytometry has been introduced in our
    laboratory since 03 years
  • we analyze             - Chronic
    lymphoproliferative disorders             - Acute
    Leukemia             - Non-Hodgkin Lymphoma on
    fine needle aspiration of               lymph
    nodes and trituration after biobsy             -
    PHN
  • Acquisition and analysis by flow cytometry
    on a BD Facs Calibur 4 colors.

4
  • Study Methods
  • - We report results of 142 cases
    of AL.
  •    - Study of the cytologic blood
    smear and bone marrow.
  •                 - Samples for CMF                
                                        -
    Blood                                             
           - Bone Marrow                -
    Immunophenotyping using a panel of monoclonal
    Antibody                  broad
    targeting                                        
          Myeloid populations                       
                            B Lymphoid
    populations

  • T Lymphoid populations
  • Classification by EGIL score to determine the
    lines B, T, myeloid and biphenotypic
  •  

5
  • Materials 142 cases
  • Sex /Age Children 44 cases (30,9)
    Sex ratio 0,83 (M 20/ W24).

  • average age 08 years (03 days - 15
    years).
  • Adults 98
    cases (69) Sex ratio 1,72 (M62/
    W36).

  • average age 43,1 years (16 82).

6
  • Study materials           
  •                1 Samples for FCM
    - Blood sample 54 cases (38)
                                                     
                             - Bone  marrow sample
    88 cases (62)
  •     2 - Filtration of Bone Marrow
  •                 3 - Count GB 94 397/mm3
    (3000 - 518 000)
  •                 4 - Dilution If GB gt 20
    000/mm3
  •                 5 - Sample preparation (FITC,
    PE, PerCP Cy5.5, APC)
  •                 6 - Acquisition
  •                 7 - Analysis - FSC/ SSC
                                         -
    Determining the negative zone                    
                      - Gating CD
    45                                      -
    Analysis of various fluorochromes positivity and
     intensity.

7
  • Results of FCM
  • 04 Groups
  • ALL 60
    cases (42,3)
  • AML 76
    cases ( 53,5)
  • Biphénotypic
    AL 06cases (4,2)
  • A L
    undifferentiated 05 cas

8
  • Results of FCM Group 01
  • ALL 60
    cases

9

  • Children ALL B 16 cases
  • Results of FCM 1
    ALL T 13 cases
  • ALL 60 cases

  • Adults ALL B 16 cases


  • AL L T 15 cases

10
  • Comparison of frequencies of different
    ALL subtypes () with literature

11
  • Percentage of ALL T in different
    countries

12
  • Results FCM 2


  • Children 12 cases (16)
  • AML 76 cases

  • Adults 64 cases (84)

13
  • AML
  • AML3 FISH t (15 17) observed
    in 03/04 studied

14
  • Frequency of different markers
    in AML

15
  • Résults of FCM 3
  • Biphénotypic LA -
    03 Children

  • - 03 Adults
  • Cases N 2 karyotype and
    FISH Philadelphia chromosome positive.          
             Cases N 5 karyotype and FISH
    trisomy 21.

16
  • FCM ALL Pre
    B

17
  • FCM AL
    L T cortical

18
  • Conclusions
  • Recently introduced into our environment,
    FCM has been a major         contribution to
    the diagnosis of AL and it usefully complements
  • the cytological and cytochemical
    traditionally existing in our
  • laboratory and was often decisive in the
    statement of the type of AL
  • and thus the therapeutic orientation.
  •  

19
  • References
  • 1 Flow Cytometry in Hematopathology A
    Visual Approach to Data Analysisand
    Interpretation Doyen
  • Nguyen, MD Lawrence W. Diamond, MD
    Raul C. Braylan, MD.
  •     2 British Committee for Standards in
    Haematology, Revised guidelines for
    immunophenotyping
  • of acute leukemia and chronic
    lymphoproliferative disorders, Clin. Lab.
    Haematol., 24, 113, 2002
  • 3 Immunophénotypage des hémopathies
    malignes Bernard Husson Centres Hospitaliers
    Jolimont
  • Lobbes Site de Jolimont Laboratoire
    de Biologie Clinique - Département Hématologie
    Unité de CMF
  • 7100 Haine-Saint-Paul.
  • 4 Hematopathology Farmarz Naeim et
    Col
  • 5 Ludwig W. D Reiter. A, Loffler.
    H, and al
  • Immunophenotypic Features of
    childhood and Adult Acute lymphoblastic Leukemia
    ( ALL)
  • Experience of the German
    Multicentre Trials ALL BFM and GMALL
  • Leukemia and lymphoma , 1994, Vol
    13, Supp 1 , pp 71 76.
  • 6 Trabzi Azeli Anissa
  • Application de limmunophénotypage à
    la classification des leucémies aigues
  • These pour le diplôme de docteur en
    science médicales
  • Juillet 1999, Faculté de Médecine
    dAlger
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