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A standardised reporting program: reporting in the 21st century
Pathology (2012) 44(S1), pp. S51–S57
DS Nelson Trainee Prize Presentation
DEVELOPMENT, VALIDATION AND IMPLEMENTATION OF 8-COLOUR PLASMA CELL FLOW CYTOMETRY AT PETER MACCALLUM CANCER CENTRE Piers Blombery, Shaun Fleming, Vuong Nguyen, Peter Gambell, Neil Came Peter MacCallum Cancer Centre, East Melbourne, Vic, Australia Aim: Multiparametric flow cytometry is an established modality in diagnostic and disease response assessment of plasma cell dyscrasias. Peter MacCallum Cancer Centre (PMCC) employed 4-colour plasma cell flow cytometry (PCFC) with 0.01% sensitivity, but the increasing clinical demand for consistently high sensitivity precipitated purchase of an 8-colour platform. Solutions to technical challenges during validation are presented. Methods: An 8-colour plasma cell (PC) antibody panel was designed from the clinical literature. Seventeen bone marrow aspirate samples were analysed concurrently on FACSCalibur and FACSCanto II instruments. Antibody titration, photomultiplier tube voltage, compensation and control testing were compared. Results: The antibody panel was revised due to influence of lysate and poor signal-to-noise ratio from auto-fluorescence of antibodies crucial to the distinction between normal and abnormal PCs (CD19, CD45, CD56). Further refinement included substituting isotype with internal controls and incorporation of additional diagnostic and prognostically relevant antibodies. The final panel demonstrated superior sensitivity, specificity, precision and accuracy to that of the established panel and additional information on the lymphopoietic compartment, of particular interest to our research clinicians. The reporting template has therefore expanded to accommodate malignant and normal PC phenotypes, lymphocyte subsets, and evidence-based interpretive comments. Conclusion: 8-colour PCFC has been validated and implemented at PMCC with subsequent improvements in diagnosis, prognostication and MRD detection. A STANDARDISED REPORTING PROGRAM: REPORTING IN THE 21ST CENTURY Travis Brown Royal Melbourne Hospital, Melbourne, Vic, Australia Ever since the division of surgery from pathology in the mid to late 19th century, the anatomical pathology report has been the primary link between pathologists and clinicians. In earlier times, the pathology report was significantly shorter and rarely longer than a few sentences in total. Nowadays, the pathology report includes five sections (patient data and history, macroscopic description, microscopic description, diagnosis and comments), ranges from paragraphs to pages and contains so much jargon that few, if any, outside the pathology field fully understand it. To add to the confusion, a single clinician can receive a multitude of reporting formats and styles for identical diagnoses simply because they were reported by different pathologists. In an effort to address these issues, the Royal College of Pathologists of Australasia have Print ISSN 0031-3025/Online ISSN 1465-3931
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developed a series of structured reporting protocols that provides guidance as to the content and presentation for cancer reports. Using these protocols, I have written a computer program that generates standardised pathology reports for pathologists to use in everyday practice. This application greatly reduces the time spent on reporting while providing important and consistent information to clinicians in accordance these recommendations. THE PROGNOSTIC SIGNIFICANCE OF THE V600E B-RAF MUTATION IN PAPILLARY THYROID CARCINOMA: DATA FROM AN AUSTRALIAN POPULATION Fiona Y. Chan, Yong Yu, Richard A. Williams, Penny A. McKelvie Department of Anatomical Pathology, St Vincent’s Hospital Melbourne, Vic, Australia Aims: B-Raf V600E mutation in the thyroid is a specific marker of papillary thyroid carcinoma (PTC) and correlates with poorer survival and rates of recurrence.1,2 We assessed the prevalence of this mutation in a series of PTCs from an Australian population, its association with aggressive clinicopathological features, recurrence and mortality. Methods: 82 patients with a median follow up of 6 years were studied. V600E mutation status was determined by PCR on DNA extracted from paraffin blocks. Clinicopathological parameters, recurrence and mortality were analysed according to V600E mutation status. Results: The V600E mutation was demonstrated in 44 of 82 (54%) tumours. Up to 58% of patients with V600E positive PTCs suffered a recurrence compared to 12% of patients with V600E negative PTCs ( p ¼ 0.005). Up to 15% of patients with V600E positive PTCs died during the follow-up period compared to 5% of patients without the mutation ( p ¼ 0.19). The V600E mutation was associated with male sex ( p ¼ 0.0002) and the classic subtype ( p ¼ 0.005). Discussion: The prevalence of B-Raf V600E mutation in PTCs in this study is similar to other series. We confirmed that this mutation was associated with a poorer prognosis, specifically higher rates of recurrence. References 1. Lupi C, Giannini R, Ugolini C, et al. Association of BRAF V600E mutation with poor clinicopathological outcomes in 500 consecutive cass of papillary thyroid cardcinoma. J Clin Endocrinol Metab 2007; 92: 4085–90. 2. Xing M, Westra WH, Tufano RP, et al. BRAF mutation predicts a poorer clinical prognosis for papillary thyroid cancer. J Clin Endocrinol Metab 2005; 90: 6373–9.
NON-DELETIONAL MUTATIONS IN PATIENTS WITH ALPHA (–a3.7 DELETION) THALASSAEMIA TRAIT: ACCURATE DIAGNOSIS AND REPRODUCTIVE IMPLICATIONS Annie Chow1, Jill Finlayson1,3, Christopher Newbound2, Laura Greenwood2, Dianne Grey1, Paula Holmes1, Michelle Jennens2, Nicole Pell2, John Beilby2,3, Reza Ghassemifar1,3 1 Department of Hematology, and 2Department of Diagnostic Molecular Biology, PathWest Laboratory Medicine, QEII Medical
2012 Royal College of Pathologists of Australasia
Copyright © Royal College of pathologists of Australasia. Unauthorized reproduction of this article is prohibited.
DS Nelson Trainee Prize Presentation
DEVELOPMENT, VALIDATION AND IMPLEMENTATION OF 8-COLOUR PLASMA CELL FLOW CYTOMETRY AT PETER MACCALLUM CANCER CENTRE Piers Blombery, Shaun Fleming, Vuong Nguyen, Peter Gambell, Neil Came Peter MacCallum Cancer Centre, East Melbourne, Vic, Australia Aim: Multiparametric flow cytometry is an established modality in diagnostic and disease response assessment of plasma cell dyscrasias. Peter MacCallum Cancer Centre (PMCC) employed 4-colour plasma cell flow cytometry (PCFC) with 0.01% sensitivity, but the increasing clinical demand for consistently high sensitivity precipitated purchase of an 8-colour platform. Solutions to technical challenges during validation are presented. Methods: An 8-colour plasma cell (PC) antibody panel was designed from the clinical literature. Seventeen bone marrow aspirate samples were analysed concurrently on FACSCalibur and FACSCanto II instruments. Antibody titration, photomultiplier tube voltage, compensation and control testing were compared. Results: The antibody panel was revised due to influence of lysate and poor signal-to-noise ratio from auto-fluorescence of antibodies crucial to the distinction between normal and abnormal PCs (CD19, CD45, CD56). Further refinement included substituting isotype with internal controls and incorporation of additional diagnostic and prognostically relevant antibodies. The final panel demonstrated superior sensitivity, specificity, precision and accuracy to that of the established panel and additional information on the lymphopoietic compartment, of particular interest to our research clinicians. The reporting template has therefore expanded to accommodate malignant and normal PC phenotypes, lymphocyte subsets, and evidence-based interpretive comments. Conclusion: 8-colour PCFC has been validated and implemented at PMCC with subsequent improvements in diagnosis, prognostication and MRD detection. A STANDARDISED REPORTING PROGRAM: REPORTING IN THE 21ST CENTURY Travis Brown Royal Melbourne Hospital, Melbourne, Vic, Australia Ever since the division of surgery from pathology in the mid to late 19th century, the anatomical pathology report has been the primary link between pathologists and clinicians. In earlier times, the pathology report was significantly shorter and rarely longer than a few sentences in total. Nowadays, the pathology report includes five sections (patient data and history, macroscopic description, microscopic description, diagnosis and comments), ranges from paragraphs to pages and contains so much jargon that few, if any, outside the pathology field fully understand it. To add to the confusion, a single clinician can receive a multitude of reporting formats and styles for identical diagnoses simply because they were reported by different pathologists. In an effort to address these issues, the Royal College of Pathologists of Australasia have Print ISSN 0031-3025/Online ISSN 1465-3931
#
developed a series of structured reporting protocols that provides guidance as to the content and presentation for cancer reports. Using these protocols, I have written a computer program that generates standardised pathology reports for pathologists to use in everyday practice. This application greatly reduces the time spent on reporting while providing important and consistent information to clinicians in accordance these recommendations. THE PROGNOSTIC SIGNIFICANCE OF THE V600E B-RAF MUTATION IN PAPILLARY THYROID CARCINOMA: DATA FROM AN AUSTRALIAN POPULATION Fiona Y. Chan, Yong Yu, Richard A. Williams, Penny A. McKelvie Department of Anatomical Pathology, St Vincent’s Hospital Melbourne, Vic, Australia Aims: B-Raf V600E mutation in the thyroid is a specific marker of papillary thyroid carcinoma (PTC) and correlates with poorer survival and rates of recurrence.1,2 We assessed the prevalence of this mutation in a series of PTCs from an Australian population, its association with aggressive clinicopathological features, recurrence and mortality. Methods: 82 patients with a median follow up of 6 years were studied. V600E mutation status was determined by PCR on DNA extracted from paraffin blocks. Clinicopathological parameters, recurrence and mortality were analysed according to V600E mutation status. Results: The V600E mutation was demonstrated in 44 of 82 (54%) tumours. Up to 58% of patients with V600E positive PTCs suffered a recurrence compared to 12% of patients with V600E negative PTCs ( p ¼ 0.005). Up to 15% of patients with V600E positive PTCs died during the follow-up period compared to 5% of patients without the mutation ( p ¼ 0.19). The V600E mutation was associated with male sex ( p ¼ 0.0002) and the classic subtype ( p ¼ 0.005). Discussion: The prevalence of B-Raf V600E mutation in PTCs in this study is similar to other series. We confirmed that this mutation was associated with a poorer prognosis, specifically higher rates of recurrence. References 1. Lupi C, Giannini R, Ugolini C, et al. Association of BRAF V600E mutation with poor clinicopathological outcomes in 500 consecutive cass of papillary thyroid cardcinoma. J Clin Endocrinol Metab 2007; 92: 4085–90. 2. Xing M, Westra WH, Tufano RP, et al. BRAF mutation predicts a poorer clinical prognosis for papillary thyroid cancer. J Clin Endocrinol Metab 2005; 90: 6373–9.
NON-DELETIONAL MUTATIONS IN PATIENTS WITH ALPHA (–a3.7 DELETION) THALASSAEMIA TRAIT: ACCURATE DIAGNOSIS AND REPRODUCTIVE IMPLICATIONS Annie Chow1, Jill Finlayson1,3, Christopher Newbound2, Laura Greenwood2, Dianne Grey1, Paula Holmes1, Michelle Jennens2, Nicole Pell2, John Beilby2,3, Reza Ghassemifar1,3 1 Department of Hematology, and 2Department of Diagnostic Molecular Biology, PathWest Laboratory Medicine, QEII Medical
2012 Royal College of Pathologists of Australasia
Copyright © Royal College of pathologists of Australasia. Unauthorized reproduction of this article is prohibited.