Dacie and Lewis Practical Haematology PDF
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1 Dacie and Lewis Practical Haematology Dacie and Lewis Practical TWELFTH EDITION Haematology BARBARA J. BAIN, MB BS, FRACP, FRCPath Professor of Diagnostic Haematology, Imperial College Faculty of Medicine, St. Mary’s Hospital, London, UK IMELDA BATES, MB BS, MD, MA, FRCPath Professor of Tropical Haematology, Liverpool School of Tropical Medicine, Liverpool, UK MICHAEL A. LAFFAN, DM, FRCP, FRCPath Professor of Haemostasis and Thrombosis, Honorary Consultant Haematologist, Imperial College Faculty of Medicine, Hammersmith Hospital, London, UK Editor Emeritus S. MITCHELL LEWIS, BSc, MD, FRCPath, DCP, FIBMS Emeritus Reader in Haematology, Imperial College Faculty of Medicine, Hammersmith Hospital, London, UK Search full text online at ExpertConsult.com © 2017, Elsevier Limited. All rights reserved. First edition 1950 Seventh edition 1991 Second edition 1956 Eighth edition 1995 Third edition 1963 Ninth edition 2001 Fourth edition 1968 Tenth edition 2006 Fifth edition 1975 Eleventh edition 2012 Sixth edition 1984 Twelfth edition 2017 Preface figure of John V. Dacie, with permission from Lewis M; Sir John Dacie MD, FRS, FRCP, FRCPath, Br J Haematol. 2005 Sep;130(6):834-44. The right of Barbara J. Bain, Imelda Bates, Michael A. Laffan and S. Mitchell Lewis, to be identified as authors of this work has been asserted by them in accordance with the Copyright, Designs and Patents Act 1988. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/permissions. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein). Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds or experiments described herein. In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. With respect to any drug or pharmaceutical products identified, readers are advised to check the most current information provided (i) on procedures featured or (ii) by the manufacturer of each product to be administered, to verify the recommended dose or formula, the method and duration of administration and contraindications. It is the responsibility of practitioners, relying on their own experience and knowledge of their patients, to make diagnoses, to determine dosages and the best treatment for each individual patient and to take all appropriate safety precautions. To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein. ISBN: 978-0-7020-6696-2 International ISBN: 978-0-7020-6930-7 eBook ISBN: 978-0-7020-6935-3 Printed in China Last digit is the print number: 9 8 7 6 5 4 3 2 1 Senior Content Strategist: Suzanne Toppy Content Development Specialist: Humayra Rahman Khan Project Manager: Joanna Souch Design: Miles Hitchen Illustration Manager: Brett MacNaughton Marketing Manager: Michele Milano Preface Sir John V. Dacie, MD, FRCPath, FRS (1912–2005). S. Mitchell Lewis, BSc, MD, DCP(London), FRCPath, FIBMS (b. 1924). This 12th edition celebrates the 66th year of Practical The 12th edition, like its predecessors, incorporates Haematology, a notable achievement. The first edition by the latest advances in laboratory haematology while JV (later Professor Sir John) Dacie was published in 1950. c ontinuing to describe traditional techniques that remain This work, and subsequent editions with Mitchell Lewis as applicable, particularly, but not only, in under-resourced co-a uthor, were based on the haematology course for the laboratories in low- and middle-income countries. University of London Diploma of Clinical Pathology and It is with sadness that we record the death of one of the subsequently the MSc in Haematology at the then Royal authors, Ms Carol Briggs, BSc FIBMS, during the prepara- Postgraduate Medical School. tion of this edition. In the last 66 years the techniques and instrumentation We are honoured to have taken over the editorship of available to the laboratory haematologist have expanded Practical Haematology from our distinguished p redecessors, at a rate once undreamed of. What has not changed is that Sir John Dacie and Dr Mitchell Lewis. We hope that our laboratory haematology continues to provide the b edrock efforts have done them justice. that supports the equally astonishing developments in clinical haematology. Haematology as a discipline remains Barbara J. Bain strongest when it is an integrated discipline with a very Michael A. Laffan close relationship between the laboratory and the c linical service. Reflecting this ideal state, the authors of t his Imelda Bates e dition include laboratory scientists and clinical and labo- ratory haematologists. vi Contributors The editor would like to acknowledge and offer grateful thanks for the input of all previous editions’ contributors, with- out whom this new edition would not have been possible. Barbara J. Bain, MB BS, FRACP, FRCPath Barbara De la Salle, MSc Professor of Diagnostic Haematology Director, UK NEQAS Haematology Centre for Haematology, Imperial College UK NEQAS Haematology and Transfusion Faculty of Medicine West Hertfordshire Hospitals NHS Trust St. Mary’s Hospital Watford, UK London, UK Letizia Foroni, MD, PhD, FRCPath Imelda Bates, MB BS, MD, MA, FRCPath Principal Teaching Fellow Professor of Tropical Haematology Centre for Haematology Liverpool School of Tropical Medicine Imperial College Faculty of Medicine Liverpool, UK Hammersmith Hospital London, UK Anne E. Bradshaw, BSc, FIBMS, DMLM Head of Operations and Regulatory Affairs G areth Gerrard, BSc, PGCert, MSc, PhD John Goldman Centre for Cellular Therapy Pathology Core Facility Manager Hammersmith Hospital U CL Cancer Institute London, UK L ondon, UK Carol Briggs,* BSc, FIBMS Previously Head of Haematology Evaluation Unit Dominic J. Harrington, MSc, PhD Department of Haematology Evaluations Consultant Clinical Scientist and Reader in University College London Hospital Diagnostic Haematology London, UK King’s College London Department of Haemostasis, Thrombosis John Burthem, PhD, FRCP, FRCPath and Nutristasis (Viapath) Clinical Senior Lecturer and Honorary Consultant Guy’s and St. Thomas’ Hospital Haematologist London, UK Department of Clinical Haematology Manchester Royal Infirmary Sandra Hing, BSc Manchester, UK Principal Molecular Geneticist Cellular and Molecular Pathology Carol Cantwell, CSci, FIBMS, DMS Imperial College Healthcare NHS Trust Previously Transfusion Laboratory Manager London, UK St Mary’s Hospital Imperial College NHS Trust Michael A. Laffan, DM, FRCP, FRCPath London, UK Professor of Haemostasis and Thrombosis and Honorary Consultant Haematologist Jane Y. Carter, MB BS, FRCPC Centre for Haematology, Imperial College Technical Director, Clinical and Diagnostics Faculty of Medicine Amref Health Africa Headquarters Hammersmith Hospital Nairobi, Kenya London, UK *Deceased vii viii Contributors Mark Layton, FRCP, FRCPH Kuldip S. Nijran, BSc, MSc, DMS, PhD, Consultant Haematologist MIPEM, CSci Imperial College Healthcare NHS Trust Head of Nuclear Medicine Physics Hammersmith and St. Mary's Hospitals Radiological Sciences Unit London, UK Imperial College Healthcare NHS Trust Hammersmith Hospital S. Mitchell Lewis, BSc, MD, FRCPath, DCP, FIBMS London, UK Emeritus Reader in Haematology Centre for Haematology, Imperial College Andrew Osei-Bimpong, MSc, CSci, FIBMS, MIHM Faculty of Medicine Laboratory Manager Hammersmith Hospital Blood Sciences London, UK Hammersmith Hospital Imperial College Healthcare NHS Trust Richard A. Manning, BSc, CSci, FIBMS London, UK Chief Biomedical Scientist Specialist Coagulation David J. Perry, MD PhD FRCPEdin FRCPLond Imperial College Healthcare NHS Trust FRCPath FAcadMEd Hammersmith Hospital Consultant Haematologist and Associate Lecturer London, UK Cambridge Haemophilia & Thrombophilia Centre Cambridge University Hospital NHS Foundation Trust Alison M. May, PhD Addenbrooke’s Hospital Previously Senior Research Fellow Cambridge UK Department of Haematology Cardiff University School of Medicine Fiona A.M. Regan, MB BS, FRCP, FRCPath Cardiff, UK Consultant Haematologist NHS Blood and Transplant (North London) Christopher McNamara, FRACP, FRCPA FRCPath and Imperial College Healthcare NHS Trust Consultant Haematologist Hammersmith Hospital Department of Haematology London, UK University College Hospital London, UK Alistair G. Reid, BSc, PhD, FRCPath Consultant Clinical Scientist Clare Milkins, BSc CSci FIBMS Cellular and Molecular Pathology Manager, UK NEQAS Blood Transfusion Laboratory Practice Imperial College Healthcare NHS Trust UK NEQAS Haematology and Transfusion London, UK West Hertfordshire Hospitals NHS Trust Watford, UK Stephen J. Richards, PhD FRCPath Consultant Clinical Scientist Ricardo Morilla, MSc, FRMS Haematological Malignancy Diagnostic Service Head of Immunophenotyping Leeds Cancer Centre Haemato-Oncology Section St James's University Hospital Royal Marsden Hospital NHS Foundation Trust Leeds, UK Sutton, Surrey, UK Lynn D. Robertson, MSc Alison M. Morilla, BSc Laboratory Manager Senior Clinical Scientist Core and Special Haematology Haemato-Oncology Section Imperial College Healthcare NHS Trust Royal Marsden Hospital NHS Foundation Trust Hammersmith Hospital Sutton, Surrey, UK London, UK Elisabet Nadal-Melsió, MD David Roper, MSc, CSci, FIBMS Consultant Haematologist Previously Principal Biomedical Scientist; Cellular and Molecular Pathology Diagnostic Haematology Imperial College Healthcare NHS Trust Imperial College Healthcare NHS Trust Hammersmith Hospital Hammersmith Hospital London, UK London, UK Contributors ix Megan Rowley, FRCP, FRCPath Barbara J. Wild, PhD, FIBMS Consultant in Haematology and Transfusion Medicine Haemoglobinopathy Specialist Consultant St. Mary’s Hospital UK NEQAS Haematology and Transfusion Imperial College Healthcare NHS Trust West Hertfordshire Hospitals NHS Trust London, UK Watford, UK Jecko Thachil, MRCP, FRCPath Nay Win, MB BS, FRCP, FRCPath, CTM(Edin) Consultant Haematologist Consultant Haematologist Haematology Department Red Cell Immunohaematology Manchester Royal Infirmary NHS Blood and Transplant (Tooting) Manchester, UK London, UK Sarmad Toma, MBChB, MSc Mark Worwood, PhD, FRCPath, FMedSci Clinical Scientist Emeritus Professor Cellular and Molecular Pathology Cardiff University School of Medicine Imperial College Healthcare NHS Trust Cardiff, UK Hammersmith Hospital London, UK 1 Collection and Handling of Blood Christopher McNamara CHAPTER OUTLINE Biohazard precautions, 1 Sample homogeneity, 4 Procurement of venous blood, 1 Serum, 4 Equipment, 1 Cold agglutinins, 4 Specimen containers, 1 Anticoagulants, 4 Phlebotomy procedure, 2 Ethylenediaminetetra-acetic acid (EDTA), 4 Postphlebotomy procedure, 3 Trisodium citrate, 4 Capillary blood, 3 Heparin, 5 Collection of capillary blood, 3 Effects of storage on the blood count, 5 Blood film preparation, 3 Effects of storage on blood cell morphology, 5 Differences between capillary and venous blood, 3 Following an informed decision to analyse a blood sam- PROCUREMENT OF VENOUS ple, a specimen must be safely and correctly procured. It is essential to be aware that variation in this pre-analytical BLOOD phase of the testing process can lead to errors in the ana- Equipment lytical phase (see Box 1-1). Venous blood is used for most examinations. Capillary It is important to assemble a tray or prepare a workspace blood samples may be satisfactory for some purposes but that has all the requirements for blood collection (Box 1-2). in general the use of capillary blood should be restricted to The selection of needle diameter is a compromise between children and to some point-of-care screening tests. achieving adequate flow with minimal turbulence and minimising patient discomfort. A 19-gauge (19G) or 21G* needle is suitable for most adults. A 23G needle is often se- lected for children. The shaft of the needle should be short BIOHAZARD PRECAUTIONS (about 15 mm). It may be helpful to collect the blood by Laboratory policies must be in place to ensure that staff means of a winged needle (often referred to as a ‘butterfly’) who collect blood samples and transfer them to the labora- connected to a length of plastic tubing that can be attached tory minimise the risk of infection from various pathogens to the nozzle of the syringe or to a needle for entering the during all aspects of specimen handling (see Chapter 24). cap of an evacuated container (see Specimen Containers). Additional precautions should be taken when handling h hiagvhin-rgi sak vsipraelc ihmaenmso (rer.hga. gthico fsev ferro)m.1 Ipna tiheinst sc isrucuspmesctaendc oef, Specimen containers the collection policy should stipulate the use of personal Containers for testing whole blood are available protective equipment, such as disposable gloves, body c ommercially with dipotassium, tripotassium or disodium apron and protective eyewear. Care must be taken to pre- vent injuries, especially when handling and disposing of *The International Organisation for Standardisation has established a needles and lancets. Recommendations for standardising standard (ISO 7864), which relates the following diameters for the blood collection have been published.2,3 different gauges: 19G = 1.1 mm; 21G = 0.8 mm; 23G = 0.6 mm. 1 2 Practical Haematology There is no universal agreement regarding the colours used BOX 1-1 Causes of misleading results for identifying containers with different additives so phle- related to specimen collection botomists should familiarise themselves with the colours PRE-COLLECTION used by their local suppliers. • Urination within 30 min; food or water intake within Evacuated tube systems in common use consist of a 2 h glass or plastic tube under a defined vacuum, a needle • Smoking and a needle holder, which secures the needle to the tube. • Physical activity (including fast walking) within 20 min The main advantage is that the cap can be pierced so that • Stress it is not necessary to remove it either to fill the tube or • Drugs or dietary supplement administration within 8 h subsequently to withdraw samples for analysis, thus min- imising the risk of aerosol discharge of the contents. An DURING COLLECTION evacuated system is useful when multiple samples in dif- • Different times (diurnal variance) ferent anticoagulants are required. The vacuum controls • Posture: lying, standing or sitting the amount of blood that enters the tube, ensuring an ad- • Haemoconcentration from prolonged tourniquet pressure equate volume for testing with the correct proportion of • Excessive negative pressure when drawing blood into any anticoagulant. syringe • Incorrect type of tube Phlebotomy procedure • Capillary versus venous blood Staff undertaking this procedure should be adequately HANDLING OF SPECIMEN trained. The phlebotomist must check that the patient’s • Insufficient or excess anticoagulant identity corresponds to the details on the request form • Inadequate mixing of blood with anticoagulant and also ensure that the phlebotomy tray contains all the • Error in patient and/or specimen identification required specimen containers and other equipment neces- • Inadequate specimen storage conditions sary for the procedure. • Delay in transit to laboratory A tourniquet should be applied just above the intended venepuncture site. Blood is best withdrawn from an ante- cubital vein or other visible veins of the forearm by means of either an evacuated tube or a syringe. It is recommended BOX 1-2 Items to be included in a that the skin be cleaned with 70% alcohol (e.g., isopro- phlebotomy tray panol) and allowed to dry spontaneously before being punctured. The tourniquet should be released as soon as • Syringes and needles the vein is punctured and blood begins to flow into the sy- • Tourniquet ringe or evacuated tube – delay in releasing the tourniquet • Specimen containers (tubes or evacuated tube system) – leads to fluid shift and haemoconcentration as a result of ve- plain and with various anticoagulants 6 • Request form nous blood stagnation. After the vein has been successfully • 70% isopropanol swabs or 0.5% chlorhexidine punctured, the piston of the syringe should be withdrawn • Sterile gauze swabs slowly with no attempt being made to withdraw blood faster • Adhesive dressings than the vein is filling. Anticoagulated specimens must be • Self-sealing plastic bags with a separate compartment mixed by inverting the container several times. The risk of for the request form unwanted haemolysis of the specimen can be minimised • Rack to hold specimens upright during process of filling by using minimal tourniquet time, withdrawing blood (except when an evacuated tube system is used) carefully, using an appropriately sized needle, delivering the • Puncture-resistant disposal container blood slowly into the receptacle and avoiding unnecessary agitation when mixing with the anticoagulant. Note that if blood is drawn too slowly or is inadequately mixed with ethylenediaminetetra-acetic acid (EDTA) anticoagulant, the anticoagulant some coagulation may occur, rendering and often have a mark to indicate the correct amount of the sample unsuitable. After collection, containers must be 4 blood to be added. Containers are also available contain- firmly capped to minimise the risk of leakage. ing trisodium citrate, heparin or acid–citrate–dextrose, as If blood collection fails, it is important to remain calm, well as containers with no additive which are used when communicate with the patient and consider the possible serum is required. Design requirements and other specifi- causes. These include poor technique (e.g., passing the cations for specimen collection containers have been de- needle through the vein, or poor selection of veins), scar- scribed in a number of national and international standards ring of tissues and haematoma formation. (e.g., that of the International Council for Standardisation After obtaining the necessary specimens, remove the 5 in Haematology ) and the European standard (EN 14820). needle and press a sterile swab over the puncture site.
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