Optimisation and validation of a PCR for antigen receptor rearrangement (PARR) assay to detect clonality in canine lymphoid malignancies
| Autor: | Elspeth M, Waugh, Alice, Gallagher, Hayley, Haining, Pamela E J, Johnston, Francesco, Marchesi, Ruth F, Jarrett, Joanna S, Morris |
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| Rok vydání: | 2016 |
| Předmět: |
Gene Rearrangement
Male Lymphoma B-Cell Genotype Lymphoma Gene Rearrangement T-Lymphocyte Lymphoma T-Cell Classification Polymerase Chain Reaction PARR Article Immunophenotyping Receptors Antigen Dogs Animals Female Dog Diseases Canine lymphoma Gene Rearrangement B-Lymphocyte Immunophenotype GeneScan DNA Primers |
| Zdroj: | Veterinary Immunology and Immunopathology |
| ISSN: | 1873-2534 |
| Popis: | Highlights • Ten primer sets detected clonality with high specificity and sensitivity. • Four extra primer sets may detect clonality in samples with equivocal results. • Knowledge of sample quality is needed for interpretation of results. • Samples generating dominant peaks require careful interpretation. 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. |
| Databáze: | OpenAIRE |
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