D.M. COURSE IN HEMATOLOGY

D.M. COURSE IN HEMATOLOGY

Course content:
Curriculum
I. Practical Laboratory Training
A. General Hematology
1. Proper use and care of common instruments such as light microscope,
centrifuge, water baths, freezers, weighing balance, etc.
2. Blood collection samples - venepuncture and finger prick methods of
sample collection, types of anticoagulants, containers and the effects of
delay in processing and storage.
3. Determination of peripheral blood counts (Hemoglobin, Hematocrit, Total
WBC and platelets) manually and calculation of red cell indices.
4. Use of automated blood cell counters including principles and practice.
Interpretation of peripheral blood counts and abnormal flags.
5. Preparation of blood films and, staining of peripheral blood films and
cytospin slides with Romanowsky and other dyes.
6. Review of normal and abnormal blood films with emphasis on morphology
of red cells, white cells and platelets.
7. Performance of WBC differential counts; subjective assessment of platelet
counts and diagnostic interpretation of abnormal counts.
8. Preparation and staining of thick and thin blood films for malarial
parasites.
9. Measurement and significance of ESR and plasma viscosity.
10. Supravital staining of reticulocytes, counting of reticulocytes.
11. Performance of bone marrow aspiration; trephine needle biopsy, splenic
aspiration.
12. Preparation of smears of bone marrow aspirates and biopsy [touch]
imprints. Staining and diagnostic evaluation of bone marrow aspirates.
13. Performance of sickling and solubility tests and screening for red cell
G6PD activity with interpretation of results.
B. Laboratory evaluation of hemolytic anemias
1. Hb S solubility test,
2. Screening for unstable hemoglobin, supravital staining of Hb H inclusions.
3. Principles and practice of separation and identification of normal and
abnormal hemoglobins by electrophoresis and chromatography,
4. Quantitation of normal and abnormal hemoglobins by densitometry and
chromatography,
5. Quantitation of HbF by alkali denaturation and cellular distribution of HbF
by Kleihauer acid elution technique,
6. Heinz body preparation,
7. Screening for G6PD deficiency and quantitative estimation of G6PD and
other red cell enzymes.
8. Direct and indirect Coomb's test, warm and cold autoantibody titres,
9. Miscellaneous biochemical test on red cell, plasma and urine,
i. Principle and procedures for estimation of plasma bilurubin and
haptoglobin; and significance of the results, screening for
methaemalbumin, methaemoglobin and sulphaehemoglobin,
ii. Screening for cryoglobins, principles of immunoglobin estimation
and immune electrophoresis,
iii. Estimation of urine for Hb, red cells, hemosiderin, urobilinogen and
bilurubin,
iv. Principles of estimation of serum ferritin and iron /TIBC,
v. Principles and estimation of red cell folate, serum folate and serumcobalamin.
C. Laboratory Investigations of Bleeding Disorders
Platelets
1) Performance of Ivy bleeding time and platelet count; study of platelet
morphology
2) Principles, practice and interpretation of platelet aggregometry tests.
3) Platelet associated immunoglobulin (PAIgG) and circulating antiplatelet
antibodies.
Screening for coagulation factor abnormalities:
1. Prothrombin time and stypven time.
2. Activated partial thromboplastin time
3. Thrombin time and reptilase time.
4. Plasma fibrinogen.
5. Correction studies with normal plasma, adsorbed plasma, aged serum and
factor deficiency plasmas.
6. FDP and D-Dimers.
7. Assays of clotting factors particularly factors VIII and IX.
8. Urea solubility test for factor XIII
9. Euglobulin Iysis time and other relevant tests of plasma fibrinolytic activity.
D Laboratory Investigation of Thrombotic Disorders
Principles:
1. Assays of plasma AT III, protein C, protein S, Factor V Leiden
2. Screening for lupus anticoagulant and activated protein C resistanceprinciples
of screening tests and interpretation of results.
3. Laboratory monitoring of anticoagulant (heparin and oral anticoagulants)
therapy.
E. Cytochemistry
Performance of the following staining procedures, Sudan Black,
Myleloperoxidase, specific and non specific esterases, acid phosphatase. PAS
and iron staining.
F. Cytogenetics
Familiarisation with cytogenetics, understanding the principles of
cytogenetics and appreciating the relevance and significance of chromosomes in
diagnostic hematology, interpreting the results of chromosome preparation of
hemopoietic cells.
G. Transfusion Medicine
1. a. ABO blood grouping (forward and backward); Rh typing (phenotypes
and genotypes), screening of antibody in sera of donors and recipients,
antibody identification following elution by various techniques.
b. Blood group compatibility (crossmatching) testing.
c. Investigation of ABO, Rh and other immunohaemolytic diseases of the
newborn.
d. Practical aspects in the selection of blood for neonatal exchange
transfusion.
2. Donor evaluation & recruitment.
3. Clinical evaluation and laboratory screening of donors to phlebotomy.
4. Phlebotomy of donors.
5. Blood component preparation and storage.
6. Practical and administrative procedures involved in issuing and
transfusing blood.
7. Principles of the mechanics of the cell separator and its use for blood
component preparation and therapeutic apheresis.
8. Practical steps in the laboratory investigation of transfusion reactions.
9. Quality control
H. Flow Cytometery
A working knowledge of the principle and practice of flowcytometery and
interpretation of the clinical significance of common leukocyte
immunophenotypes.
I Laboratory Equipment and Organization
A working knowledge of the mechanics of the various laboratory
instruments including their operation, calibration and basic maintenance, is
desirable.
1. Laboratory space distribution; ordering, siting and installation of laboratory
equipment; workflow procedures and handling of samples.
2. Staffing - technical and non-technical.
3. Use of computers and generation of laboratory statistics.
4. Health and Safety measures.[Physical/Chemical/Biological/Radiation]
5. Waste disposal.
6. Quality Assurance (Internal and External) measures.
7. Budgeting
II. Histopathology Module
Practical laboratory training and related theory should cover the following
areas:
1. General processing of tissues, especially Lymph Node Biopsy, Bone Marrow
trephine biopsy, F.N.A.C and other tissues with special emphasis on
leukemic deposits of lymphomatous involvement.
2. Techniques of cytology including cytospins in relation to body fluid of patients
with hematological disorders.
3. Immunocytochemistry relevant to Hematology.
4. Electron microscopy of hemopoietic cells.
III Biochemistry Module
A. Laboratory Techniques :
Practical "hands -on" experience and related theoretical background in the
following:-
1. Separation procedures - electrophoretic techniques, chromatography
2. Immunochemical methods.
3. Radioimmunoassays.
IV HLA Module
Demonstration and understanding the principles of:-
1. Separation of lymphocytes using density gradient and centrifugation.
2. The microlymphocytoxicity test and its application in HLA typing,
crossmatching and antibody screening.
3. The mixed lymphocyte reaction: one way and two-way cultures.
4. Miscellaneous investigations (on request) including mitogen and antigen
induced lymphocyte transformation and neutrophil function assays.
V. Molecular Biology Module
Understanding the principles involved in the molecular diagnosis of
hematological disorders.
1. DNA separation
2. PCR
3. RFLP and Southern Blotting.
II. Clinical Haematology Training
With appropriate guidance and under supervision, the post graduate
trainee will be responsible primarily for the acquisition of knowledge in all areas
of hematology and Transfusion Medicine. Such knowledge will be acquired and
demonstrated through seminars, case presentations, journal clubs, tutorials and
proper use of library for suggested reading and formal reviews of major selected
topics. Faculty staff should be present at these various exercises so as to
provide the appropriate inputs. When necessary faculty staff may be required to
review certain subjects in form of formal lectures. Clinical experience will be
acquired by the trainee by day to day management of all patients admitted to the
hematology service. Faculty will also be involved in teaching of trainees in the
ward-rounds and outpatient clinics.