Lymphoma immunophenotype of dogs determined by immunohistochemistry, flow cytometry, and polymerase chain reaction for antigen receptor rearrangements

J Vet Intern Med. 2013 Nov-Dec;27(6):1509-16. doi: 10.1111/jvim.12185. Epub 2013 Sep 20.


Background: Immunohistochemistry (IHC), flow cytometry (FC), and PCR for antigen receptor rearrangements (PARR) are 3 widely utilized tests to determine immunophenotype in dogs with lymphoma (LSA).

Objectives: This study evaluated the ability of FC and PARR to correctly predict immunophenotype as defined by IHC and to determine the level of agreement among the 3 tests.

Animals: Sixty-two dogs with lymphoma.

Methods: Retrospective study. Medical records were searched to identify dogs with LSA that had concurrent IHC, FC, and PARR performed. Immunophenotype results were categorized as B-cell, T-cell, dual immunophenotype (B- and T-cell), or indeterminate. The results of FC and PARR were evaluated for correctly classifying B- and T-cell LSA as compared with IHC. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were evaluated in addition to concordance between each test.

Results: The sensitivity of FC was significantly higher than PARR for both B-cell (91% versus 67%; P < 0.0072) and T-cell (100% versus 75%; P < 0.0312) LSA. The percent agreement between FC and IHC was 94%, between PARR and IHC was 69%, between FC and PARR was 63%, and among all 3 tests was 63%.

Conclusions and clinical importance: Flow cytometry is superior to PARR in correctly predicting immunophenotype when evaluating lymph nodes from dogs already diagnosed with B- or T-cell LSA. If fresh samples are not available for FC, PARR is an acceptable assay for determination of immunophenotype given its high specificity.

Keywords: Cancer; Flow cytometry; Lineage; Lymph nodes; Specificity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Area Under Curve
  • DNA / chemistry
  • DNA / genetics
  • Dog Diseases / immunology*
  • Dogs
  • Female
  • Flow Cytometry / veterinary
  • Immunohistochemistry / veterinary
  • Immunophenotyping / methods
  • Immunophenotyping / veterinary*
  • Lymph Nodes / immunology*
  • Lymphoma / genetics
  • Lymphoma / immunology*
  • Male
  • Polymerase Chain Reaction / veterinary
  • Receptors, Antigen / genetics
  • Receptors, Antigen / immunology*
  • Reproducibility of Results
  • Retrospective Studies
  • Sensitivity and Specificity


  • Receptors, Antigen
  • DNA