Optimisation and validation of a PCR for Antigen Receptor Rearrangement (PARR) assay to detect clonality in canine lymphoid malignancies

Waugh, E. M., Gallagher, A., Haining, H., Johnston, P. E.J., Marchesi, F. , Jarrett, R. F. and Morris, J. S. (2016) Optimisation and validation of a PCR for Antigen Receptor Rearrangement (PARR) assay to detect clonality in canine lymphoid malignancies. Veterinary Immunology and Immunopathology, 182, pp. 115-124. (doi: 10.1016/j.vetimm.2016.10.008) (PMID:27863542) (PMCID:PMC5119497)

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Abstract

PCR for antigen receptor gene rearrangements (PARR) analysis is being increasingly used to assist diagnosis of canine lymphoma. In this study, PARR was carried out on consecutive samples received as part of routine diagnostic practice from 271 patients: 195 with lymphoid malignancies, 53 with reactive conditions and 23 with other neoplasms. Initially, published primer sets were used but later minor primer modifications were introduced and primers were rationalised to give a PARR panel that provides a good compromise between sensitivity and cost. Results were compared to diagnoses made by histology or cytology, coupled with immunophenotyping by flow cytometry or immunohistochemistry where possible. After exclusion of 11 poor quality samples, 230/260 (88%) gave a clear result with 162/163 (99%) of samples classified as clonal and 56/67 (84%) classified as polyclonal giving results concordant with the cytological/histological diagnosis. Among 30 samples with equivocal results, 21 had clonal peaks in a polyclonal background and nine showed little amplification. These were from patients with a range of neoplastic and non-neoplastic conditions emphasising the need to interpret such results carefully in concert with other diagnostic tests. The combination of primer sets used in this study resulted in a robust, highly specific and sensitive assay for detecting clonality.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Jarrett, Professor Ruth and Morris, Professor Joanna and Gallagher, Dr Alice and Johnston, Dr Pamela and Marchesi, Dr Francesco and Haining, Dr Hayley and Waugh, Ms Elspeth
Authors: Waugh, E. M., Gallagher, A., Haining, H., Johnston, P. E.J., Marchesi, F., Jarrett, R. F., and Morris, J. S.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Medical Veterinary and Life Sciences > School of Veterinary Medicine
Journal Name:Veterinary Immunology and Immunopathology
Publisher:Elsevier
ISSN:0165-2427
ISSN (Online):1873-2534
Published Online:19 October 2016
Copyright Holders:Copyright © 2016 Elsevier
First Published:First published in Veterinary Immunology and Immunopathology 182:115-124
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
500191BBSRC Doctoral Training Grant 2009-15Massimo PalmariniBiotechnology and Biological Sciences Research Council (BBSRC)BB/F016786/1MVLS III - CENTRE FOR VIRUS RESEARCH