title: Optimisation and validation of a PCR for antigen receptor rearrangement (PARR) assay to detect clonality in canine lymphoid malignancies.
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.
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, 464 Bearsden Road, Glasgow, G61 1QH, UK. Electronic address: e.waugh.1@research.gla.ac.uk.
authors.forename: Elspeth M
authors.initials: EM
authors.lastname: Waugh
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, 464 Bearsden Road, Glasgow, G61 1QH, UK.
authors.forename: Alice
authors.initials: A
authors.lastname: Gallagher
authors.affiliation: School of Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, 464 Bearsden Road, Glasgow, G61 1QH, UK.
authors.forename: Hayley
authors.initials: H
authors.lastname: Haining
authors.affiliation: School of Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, 464 Bearsden Road, Glasgow, G61 1QH, UK.
authors.forename: Pamela E J
authors.initials: PEJ
authors.lastname: Johnston
authors.affiliation: School of Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, 464 Bearsden Road, Glasgow, G61 1QH, UK.
authors.forename: Francesco
authors.initials: F
authors.lastname: Marchesi
authors.affiliation: MRC-University of Glasgow Centre for Virus Research, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, 464 Bearsden Road, Glasgow, G61 1QH, UK.
authors.forename: Ruth F
authors.initials: RF
authors.lastname: Jarrett
authors.affiliation: School of Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, 464 Bearsden Road, Glasgow, G61 1QH, UK.
authors.forename: Joanna S
authors.initials: JS
authors.lastname: Morris
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004283:Dog Diseases / Q000175:diagnosis / Q000235:genetics* / Q000276:immunology*; D004285:Dogs; D005260:Female; D015321:Gene Rearrangement; D015322:Gene Rearrangement, B-Lymphocyte; D015329:Gene Rearrangement, T-Lymphocyte; D005838:Genotype; D016130:Immunophenotyping; D008223:Lymphoma / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000276:immunology / Q000662:veterinary; D016399:Lymphoma, T-Cell / Q000235:genetics / Q000276:immunology / Q000662:veterinary; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods / Q000706:statistics & numerical data / Q000662:veterinary*; D011946:Receptors, Antigen / Q000235:genetics*
chemical_list: D017931:DNA Primers; D011946:Receptors, Antigen
keywords: Canine lymphoma; Classification; GeneScan; Genotype; Immunophenotype; PARR