HAEMATOLOGY LABORATORY Head of Unit : Dr Raudhawati Osman Phone : +603-26155281 Email : raudhaosman@yahoo.com 1. INTRODUCTION The Haematology Unit, Department of Pathology, HKL provides tertiary diagnostic and consultative haematology services to all government hospitals and clinics in Malaysia. It also serves as a training centre in laboratory haematology for undergraduates from local colleges and universities as well as doctors majoring in haematology under the Masters in Pathology Program (MPATH). 2. LIST OF SERVICES AND LABORATORY TURN AROUNDTIME (LTAT) Since the department reorganization in 2006, the Haematology Unit only provides specialized haematology tests as listed below: Section Test LTAT ( working days) Morphology Full blood picture (FBP) Urgent: Routine: 10 days Bone marrow aspirate (BMA) 5 days Trephine biopsy 20 days Urine hemosiderin CSF for blast NAP score/ LAP score H inclusion Red Cell Haemoglobin analysis 20 days Disorder Hb F quantitation 5 days Heinz bodies Kleihauer test Sickling test Osmotic fragility test (OFT) Cryoglobulin 5 days Methemoglobin Flow Leukaemia/lymphoma immunophenotyping (IPT) Verbal: 36 hours
cytometry Molecular haematology Stem Cell Transplant CD4/CD8 enumeration Paroxysmal nocturnal hemoglobinuria (PNH) Immunophenotyping (IPT) Molecular alpha thalasaemia Molecular detection of mutations in acute leukaemia Molecular detection of BCR-ABL Chimerism assay CD34 enumeration Buffy coat preparation Red cell depletion Cryopreservation Donor lymphocyte cryopreservation Stem cell/ donor lymphocyte infusion (DLI) Plasma depletion Formal: 10 days 7 days 10 days 45 days 14 days 14 days 30 days 24 hours 6 hours 6 hours 24 hours 24 hours 30 minutes 4 hours 3. REQUEST FORM 3.1. All specimens must be accompanied with PER-PAT 301 form. 3.2. Forms must be filled LEGIBLY and COMPLETELY with the following information: Patient s details: Name, IC number, sex, age, ward / hospital name Patient s clinical and test details: relevant clinical history, diagnosis, tests required, type of sample, time and date of sampling, Requesting doctors details: name, stamp, signature 3.3. Incompletely filled forms may result in sample being rejected for testing. 4. SPECIMEN CONTAINERS Specimen containers used in haematology laboratory are listed below: Specimen containers EDTA tubes Lithium heparin Sodium citrate Urine container Blood bag with ACD Cryoshipper Sterile bottle Tests Generally most of the specialized haematology tests offered at our laboratory use this tube unless stated otherwise OFT tests FBP in cases of platelet clumping TRO EDTA-induced platelet clumping Urine hemosiderin Stem cell collection from bone marrow, cord blood and peripheral blood, Buffy coat preparation, Cryopreservation, Donor lymphocyte cryopreservation, plasma depletion, red cell depletion, Stem cell/ donor lymphocyte infusion Body fluids
Specimen containers except blood bag with ACD and cryoshipper can be obtained from the counter for PembekalanTiub Setempat located at the ground floor of Pathology Department HKL. 5. SPECIMEN COLLECTION 5.1. General 5.1.1. Most of the tests use venous blood samples. 5.1.2. To ensure consistent and accurate result, follow strictly to the volume of blood required for type of test specified on the label or fill the blood sample to the mark on the label. 5.1.3. To prevent haemolysis: Avoid vigorous mixing Send the specimen to the laboratory as soon as possible 5.1.4. Avoid clot formation by: Ensuring a smooth venipuncture and steady flow of blood into the syringe. Introducing the blood in the anticoagulant bottle as soon as the blood has been drawn. Immediately gently mix by inverting the tube at least 6-10 times. 5.2. Method for collection for special hematology test: 5.2.1. Full Blood Picture (FBP) Collect 2.5 ml of blood in EDTA container or fill up to the mark as instructed Mix gently to ensure good anticoagulation Send to the laboratory as soon as possible (within 4 hours of collection) 5.2.2. Bone Marrow Aspirate (BMA) Please call 6549 to arrange for an appointment On the appointment date, laboratory technologist will be at the procedure room to prepare smears from bone marrow sample obtained by ward doctor. The ward staff has to ensure that all necessary preparation is ready to avoid unnecessary waiting of the laboratory technologist prior to commencement of the procedure. Waiting time more than 15 minutes may result in the procedure been postponed. MLT in charge will provide the slides for BMA and universal container with 10% formalin for trephine biopsy on the day of the procedure. Other containers for other tests such as EDTA for immunophenotyping and molecular study, lithium heparin for cytogenetic study and blood culture bottles for marrow culture must be provided by ward staff if required.
The slides will be taken back to the haematology laboratory by the laboratory technologist for staining and reporting. 5.2.3. Haemoglobin analysis Collect 2.5 ml of blood in EDTA container Mix well to avoid clot formation Send the collected samples to the lab for further processes 5.2.4. Osmotic Fragility Test (OFT) By appointment only Collect 5.0 ml of blood into Lithium Heparin container Mix well by inverting gently the container few times Send to the laboratory before 9.00 am Test must be performed within 2 hours after blood collection if samples are kept at room temperature or within 6 hours if samples are kept at 2 0-6 0 C. 5.2.5. Test for unstable haemoglobin By appointment only Collect 2.5 ml of blood in EDTA container. Mix well and send to the laboratory before 9.00 am 5.2.6. Methaemoglobin By appointment only Collect 2.5 ml of blood in EDTA container. Mix well and send to the laboratory before 9.00 am Test must be performed within 1 hour of blood collection 5.2.7. Immunophenotyping for leukaemia/lymphoma Collect 2.5 ml of blood or bone marrow in an EDTA tube. Mix well and send to the laboratory immediately. Body fluids samples are collected in sterile bottle. Body fluids must be processed within 4 hours of collection. Peripheral blood and bone marrow specimens kept more than 48 hours are not suitable for analysis 5.2.8. IPT for PNH Collect 2.5 ml of blood in an EDTA tube Mix well and send to the laboratory immediately. Specimens kept more than 48 hours are not suitable for analysis 5.2.9. CD4/CD8 enumeration test Collect 2.5ml of blood in EDTA container or fill to the mark as instructed Mix gently by inverting the tube at least 6-10 times.
Specimens kept more than 48 hours are not suitable for analysis 5.2.10. CD34 enumeration ( PBSC, Bone Marrow and cord blood) Collect 2.5ml of blood in EDTA container or fill to the mark as instructed Mix gently by inverting the tube at least 6-10 times. Send the sample to the laboratory as soon as possible. 5.2.11. Stem cell processing and cryopreservation (PBSC/bone marrow/ cord blood/ donor lymphocyte) Prior arrangement (at least 2 weeks earlier) has to be made with pathologist (ext 5598/ 7514) or MLT in-charge (ext 6883/5711) Collect bone marrow/ cord blood/ peripheral blood stem cell (PBSC) in an appropriate blood bag and send to BMT laboratory. 5.2.12. NAP score Sample: smear from finger prick blood sample or 2.5 ml blood sample in EDTA tube Send the smear or EDTA sample to the laboratory as soon as possible. EDTA sample need to be processed within 30 minutes of blood collection. 5.2.13. CSF for blast Sample is collected in sterile bottle Sample need to be processed within 2-4 hours of sample collection 5.2.14. Molecular for alpha thalassaemia Collect 2.5 ml of blood in an EDTA tube Latest FBC and Hb analysis result must be attached with the request form Detail history on index case must be provided for family screening 5.2.15. Molecular detection of mutations in acute leukaemia and BCR-ABL for CML cases Blood or bone marrow sample is collected in EDTA tube Sample need to be processed within 4-6 hours of blood collection SUMMARY OF HAEMATOLOGY TESTS ACCORDING TO ALPHABETICAL ORDER BIL TEST CONTAINER SPECIMEN TYPE REMARKS 1 BMA Smear Bone Marrow By appointment 2 BM trephine biopsy Universal container Trephine biopsy (with 10% formalin) 3 Buffy Coat Blood bag with ACD Bone marrow harvest By appointment Preparation 4 FBP EDTA Blood 5 CD4/CD 8 EDTA Blood
enumeration 6 CD 34 enumeration EDTA Blood 7 Chimerism assay EDTA Blood/ PBSC/ bone marrow For paediatric cases only 8 Cryopreservation Blood bag with ACD PBSC/ bone marrow/ By appointment cord blood 9 CSF for blast Sterile container CSF 10 Cryoglobulin 2 EDTA tubes and 2 Blood By appointment plain tubes 11 Donor Lymphocyte Blood bag with ACD PBSC By appointment Cryopreservation 12 H inclusion EDTA Blood Reported with FBP 13 Hb analysis EDTA Blood 14 Hb F quantitation EDTA Blood 15 Heinz Bodies EDTA Blood 16 IPT for leukaemia and Lymphoma EDTA Bone marrow/ blood/ body 17 IPT for PNH 18 Kleihauer Test EDTA Blood By appointment 19 Methaemoglobin EDTA Blood By appointment 20 Molecular Alpha EDTA Blood thalasaemia 21 Molecular detection of mutations in acute leukaemia EDTA Blood/ bone marrow 22 Molecular detection of BCR-ABL EDTA Blood/ bone marrow 23 NAP Score EDTA Blood 24 OFT Lithium heparin Blood By appointment 25 Plasma depletion Blood bag with ACD bone marrow harvest By appointment 26 Red Cell Depletion Blood bag with ACD bone marrow harvest By appointment 27 Sickling Test By appointment 28 Stem Cell/Donor Cryoshipper Cryopreserved product By appointment Lymphocyte Infusion 29 Unstable EDTA By appointment Haemoglobin 30 Urine Hemosiderin Universal container 6. REJECTION CRITERIA No Reasons for rejection 1 Incorrect specimen for test requested 2 Patient name and ID on specimen container does not match information in request form 3 Incorrect container / preservative used
4 Blood clotted 5 Insufficient sample 6 Blood markedly lysed 7 Specimen spilled / leaked 8 No sample 7 Empty container 8 Broken container 9 Expired sample container 10 Lipaemic sample 11 Unlabelled specimen 12 Unsuitable sample for analysis 13 Date of sample collection is not stated 14 Test is not offered in Haematology Laboratory, HKL 15 Incomplete request form : IC number- not stated/ incomplete 16 Incomplete request form : test request- not stated/ incomplete 17 Incomplete request form : no hospital/ ward/ clinic stated 18 Incomplete request form: the name of requesting doctor is not stated. 19 Incomplete request form: no clinical history provided 20 Post-transfusion sample for hb analysis test 21 Redundant request 22 No FBC or Hb analysis results attached for molecular alpha thalassaemia test 7. SAMPLE RECEPTION 7.1. During office hour : Receiving Counter, Haematology Laboratory 7.2. After office hour : Receiving counter, Pre-Analytical Unit At the reception counter, ward or clinic staff must ensure the date and the time of sample arrival is recorded by using the electronic time clock/ recorder. 8. REPORTING OF RESULTS 8.1. The wards that have Lab viewer system can trace their result electronically. 8.2. Verbal report: will be issued for certain cases of IPT for leukaemia/ lymphoma and urgent FBP (if requested by clinicians). Printed result will be dispatched once available. 8.3. Formal report will be printed and dispatched once available. 9. COLLECTION OF RESULTS 9.1. Formal reports for in-patients and cases from Klang Valley are dispatched to designated pigeon holes located at the main counter of the department.
9.2. Formal reports for outside Klang Valley are mailed via Pos Malaysia to mailing centres (state hospitals) or specific mailing address agreed between requester and our laboratory6. 9.3. Urgent results will be informed to the wards by telephone or to be given by hand to the waiting ward personnel. 10. ENQUIRIES Enquiries regarding the haematology test, samples and result can be made via the following numbers: Section Extension Morphology 6549 Red Cell Disorder 6374 Flow cytometry 6853 Stem Cell Transplant 6883 Molecular haematology 5748/ 5746 Medical Officer Urgent FBP MO Room Pathologist Room Main laboratory Diagnostic Room 6374 7514 5598 7514 11. SERVICES AFTER OFFICE HOUR The specialized haematology laboratory does not operate after office hours. However, urgent requests for FBP and leukemia immunophenotyping can be arranged via the Medical Officer or Pathologist on-call. Please refer to the monthly roster provided by the Department of Pathology. 12. LIST OF PATHOLOGISTS AND CONTACT DETAILS Pathologists Email Ext. number Dr Raudhawati Osman raudhaosman@yahoo.com 5281 Dr Mimi Azura Aziz mimiazizmy@yahoo.com 7514/ 5598 Dr SitiShahrumMuhamed Said drsitishahrum@yahoo.com 7514/ 5598
Dr Noraesah Mahmud noraesah@yahoo.com 7514/ 5598 Dr SuzanaZainol dr_suzana_zainol@yahoo.com 7514/ 5598 Dr Thatcheiany A/P Kumariah thatcheianykumariah@yahoo.com 7514/ 5598