for Leucocyte Immunophenotyping Leukaemia Diagnosis Interpretation All Participants Date Issued: 08-September-2014 Closing Date: 26-September-2014

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1 for Leucocyte Immunophenotyping Leukaemia Interpretation All Participants Participant: 4xxxx Trial No: Date Issued: 08-September-2014 Closing Date: 26-September-2014 Trial Comments This was an electronic only trial issued to a total of 676 participants Your : Consensus : Your is classed as a CORRECT result Performance Score: Running Score: 0 Consensus Lineage Subclassification Chronic lymphocytic leukaemia/small lymphocytic lymphoma Chronic lymphocytic leukaemia/small lymphocytic lymphoma 0 Consensus Lymphoid-B cell Mature (peripheral) B-cell neoplasms Chronic lymphocytic leukaemia/small lymphocytic lymphoma Breakdown of returned diagnoses Lymphoid-B cell Your Mature (peripheral) B-cell neoplasms Chronic lymphocytic leukaemia/small lymphocytic lymphoma No Of Returns Percentage Of Returns Total Returns Chronic lymphocytic leukaemia/small lymphocytic lymphoma B-cell prolymphocytic leukaemia Plasma cell myeloma Issue Date: 07 Nov 2014 Page 1 of 7 Clinical Pathology Accreditation (UK) Ltd. Accredited EQA Schemes Copyright Sheffield Teaching Hospitals NHS Foundation Trust 2014 Not to be reproduced in all or part without permission of STH or UK NEQAS LI

2 Leukaemia Diagnostic Interpretation trial Case History A 60 year old female was seen by her General Practitioner and stated that she was experiencing tiredness, weight loss and recurrent infections. Haematological examination revealed the following: Hb- 96 g/l; WBC- 250 x10^9/l; PLT- 64 x10^9/l; RBC x10^12/l; HCT L/L; MCV fl; MCH pg Immunophenotype of the malignant/abnormal cell population The phenotype provided for this exercise was (overall median percentage values derived from part 1 given in parentheses): Positive antigens: CD5 + CD19 + CD23 + CD43 + CD200 + HLADR + Kappa + (and CD45 + ) CD5 (95%), CD19 (95%), CD23 (30%), CD43 (97%), CD45 (100%), CD200 (93%), HLADR (96%), Kappa (69%) Negative antigens: CD2 (1%), CD3 (1%), CD7 (0%), CD10 (0%), CD11c (15%), CD13 (0%), CD20 (18%), CD22 (12%), CD25 (0%), CD34 (0%), CD38 (8%), CD79b (4%), CD103 (0%), FMC7 (0%), Lambda (0%) Cytogenetics and Molecular genetics: FISH analysis of 100 interphase nuclei has shown no evidence of deletion of TP53 at 17p13.1 Issue Date: 7 Nov 2014 Page 2 of 7

3 Peripheral Blood Morphology Morphology comments from Professor Wendy Erber Consultant Haematologist, University of Western Australia, Nedlands, Australia Comment: There is a peripheral blood lymphocytosis. The majority (approximately 80%) are small round cells with a high nuclear:cytoplasmic and coarse chromatin. The remaining 20% of cells are larger with a lower nuclear:cytoplasmic ratio, less nuclear chromatin condensation and a single prominent nucleolus. Smears cells are present as well as they are being left shift. : Chronic Lymphocytic Leukaemia (CLL) with 20% prolymphocytes. Example of Digital Images Issued Figure 1: Peripheral Blood x50 magnification (May Grünwald Giesma stain) Issue Date: 7 Nov 2014 Page 3 of 7

4 Exercise Conclusion / Case Discussion Please note that, because this trial requires a diagnosis based on the WHO guidelines, this report is based on the WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4 th Edition 1. Leukaemia Immunophenotyping - Part one Please note that part 1 is considered to be a technical exercise. Several antigens on this exercise showed a fluorochrome effect with regards to the returned results, these were CD11c, CD20, CD22, CD23, CD38 and CD79b. In all of these cases the use of FITC resulted in a significant reduction in percentage cell expression (as measured by the robust mean). We would remind laboratories again that caution should be used when using FITC conjugated antibodies to detect antigens that may have weaker than normal expression and consider changing the fluorochrome to one that will increase the signal to noise ratio. Leukaemia Diagnostic Interpretation - Part Two EXERCISE CONSENSUS DIAGNOSIS (Classified as Correct) Chronic Lymphocytic Leukaemia/Small Lymphocytic Lymphoma (CLL/SLL) Other Differential Diagnoses (Classified as Major Errors): B-cell Prolymphocytic leukaemia Plasma Cell Myeloma Other Differential Diagnoses (Classified as Minor Errors): The classification of major and minor errors is currently under review and will be discussed with our steering committee in the near future. Issue Date: 7 Nov 2014 Page 4 of 7

5 Rationale for Correct Classification The immunophenotypic profile of the case combined with the morphology supports the diagnosis of CLL/SLL. The major errors are those of B-cell Prolymphocytic Leukaemia (B-PLL) and Plasma Cell Myeloma. Both being either morphologically of phenotypically distinct. It should be noted that to be defined as a B-cell Prolymphocytic Leukaemia there must be >55% prolymphocytes in the peripheral blood. Furthermore, CD5 and CD23 are only present in 20-30% of cases. Whilst this case was CD5 and CD23 positive the prolymphocytes were only 20%. B-PLL is rare, comprising approximately 1% of lymphocytic leukaemias. Plasma cell myeloma is BM-based multifocal plasma cell neoplasm. The sample issued was peripheral blood. Additionally, if the cells in this did have the phenotype of plasma cells (which they did not) then this case would have been a plasma cell leukaemia, not plasma cell myeloma. Issue Date: 7 Nov 2014 Page 5 of 7

6 Further Trial Findings Table 1: Submissions from all participants - laboratories and individuals. Total Lab Individual Chronic Lymphocytic Leukaemia/Small Lymphocytic Lymphoma B-cell Prolymphocytic Leukaemia Plasma Cell Myeloma Table 2: Submissions from laboratory participants only Lab Chronic lymphocytic leukaemia/small Lymphocytic Lymphoma 99.0% B-cell Prolymphocytic Leukaemia 1.0% Table 3: Submissions from individual participants only Individual Chronic Lymphocytic Leukaemia/Small Lymphocytic Lymphoma 94.6% B-cell Prolymphocytic leukaemia 2.7% Plasma Cell Myeloma 2.7% References: 1. Steven H. Swerdlow, Elias Campo, Nancy Lee Harris, Elaine S Jaffe, S. Jaffe, Stefano A. Pileri, Harald Stein, Jürgen Thiele, James W. Vardiman. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4 th Edition. IARC Press 2008 Issue Date: 7 Nov 2014 Page 6 of 7

7 Information with respect to compliance with standards BS EN ISO/IEC 17043: a) The proficiency testing provider for this programme is: UK NEQAS for Leucocyte Immunophenotyping Pegasus House, 4 th Floor Suite 463A Glossop Road Sheffield, S10 2QD United Kingdom Tel: +44 (0) , Fax: +44 (0) [email protected] b) The coordinators of UK NEQAS LI programmes are Prof David Barnett and Mr Liam Whitby c) Person(s) authorizing this report: Prof David Barnett, Director or Mr Liam Whitby, Operations Manager of UK NEQAS LI d) No activities in relation to this EQA exercise were subcontracted g) The UK NEQAS LI Confidentiality Policy can be found in the Quality Manual which is available by contacting the UK NEQAS LI office. Participant details, their results and their performance data remain confidential unless revealed to the relevant NQAAP when a UK participant is identified as having performance issues i) All EQA samples are prepared in accordance with strict Standard Operational Procedures by trained personnel proven to ensure homogeneity and stability. Where appropriate/possible EQA samples are tested prior to issue. Where the sample(s) issued is stabilised blood or platelets, pre and post stability testing will have proved sample suitability prior to issue l), n), o), r) & s) Please refer to the UK NEQAS LI website at for detailed information on each programme including the scoring systems applied to assess performance. Where a scoring system refers to the consensus result this means the result reported by the majority of participants for that trial issue. Advice on the interpretation of statistical analyses and the criteria on which performance is measured is also given. Please note that where different methods/procedures are used by different groups of participants these may be displayed within your report, but the same scoring system is applied to all participants irrespective of method/procedure used m) We do not assign values against reference materials or calibrants q) Details of the programme designs as authorized by The Steering Committee and Specialist Advisory Group can be found on our website at The proposed trial issue schedule for each programme is also available t) If you would like to discuss the outcomes of this trial issue, please contact UK NEQAS LI using the contact details provided. Issue Date: 7 Nov 2014 Page 7 of 7