Flow cytometric evaluation of peripheral blood and bone marrow and fine-needle aspirate samples from multiple sites in dogs with multicentric lymphoma

Joetzke, Alexa E; Eberle, N.; Nolte, Ingo; Mischke, R.; Simón, Daniela

doi:10.2460/ajvr.73.6.884

Cited by 17 publications

(16 citation statements)

References 42 publications

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“…Also, this study only evaluated the use of FC and PARR on samples from lymph nodes. Although the general sensitivity of PARR (regardless of immunophenotype) has not yet been reported using samples exclusively from extranodal sites, only 70% of extranodal sites evaluated with FC tested positive in confirmed cases of LSA in a recent study . Another limitation of this study is that all testing was performed at only 1 institution (North Carolina State University).…”

Section: Discussionmentioning

confidence: 83%

“…Although the general sensitivity of PARR (regardless of immunophenotype) has not yet been reported using samples exclusively from extranodal sites, only 70% of extranodal sites evaluated with FC tested positive in confirmed cases of LSA in a recent study. 33 Another limitation of this study is that all testing was performed at only 1 institution (North Carolina State University). Interpretation of each assay (particularly FC and PARR) is highly dependent on the skill and experience of the operator and it is possible that different results could be obtained if testing was performed elsewhere.…”

Section: Discussionmentioning

confidence: 99%

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Lymphoma Immunophenotype of Dogs Determined by Immunohistochemistry, Flow Cytometry, and Polymerase Chain Reaction for Antigen Receptor Rearrangements

Thalheim

Williams

Borst

et al. 2013

Veterinary Internal Medicne

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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.

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Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Lymphoma Immunophenotype of Dogs Determined by Immunohistochemistry, Flow Cytometry, and Polymerase Chain Reaction for Antigen Receptor Rearrangements

Thalheim

Williams

Borst

et al. 2013

Veterinary Internal Medicne

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“…When comparing different studies on canine lymphoma, various combinations of conventional and additional diagnostic techniques including abdominal ultrasonography, bone marrow aspiration cytology, immunophenotyping, buffy coat analysis and splenic or hepatic cytology can be found …”

Section: Discussionmentioning

confidence: 99%

“…Cytologic samples of liver and spleen were considered as positive for lymphoma involvement if more than 5% lymphoblasts were found . Bone marrow cytology was recorded as positive if more than 2% lymphoblasts or aggregates of lymphoblasts were observed …”

Section: Methodsmentioning

confidence: 99%

Splenic and hepatic ultrasound and cytology in canine lymphoma: effects of findings on stage migration and assessment of prognosis

Nerschbach

Eberle

Joetzke

et al. 2014

Vet Comparative Oncology

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Stage migration is described in humans and dogs as a sequel of using more sensitive diagnostic methods. One hundred eighty-six dogs with multicentric lymphoma were enrolled with results of conventional staging as well as ultrasonographic and cytological examination of liver and spleen being available. The addition of splenic respective hepatic ultrasound and cytology findings resulted in slightly lower number of dogs classified as having liver and spleen involvement. In dogs with multicentric lymphoma, addition of cytology led to a significant shift of individuals from stage IV to stage III. Findings of hepatic and splenic ultrasound and cytology exerted no significant influence on complete remission and survival durations in dogs with combination chemotherapy. Staging methodology in canine lymphoma should be redefined, considering that the prognostic significance of splenic and hepatic ultrasound and cytology warrants further investigation.

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“…Six of the dogs were classified as clinical stage III, 21 dogs as stage IV and 13 dogs as stage V. Fifteen dogs had substage a and 25 dogs had substage b. Immunophenotyping using flow cytometric analysis of lymph node aspirates was performed as described previously [18] and revealed lymphoma of the B and T cell phenotype in 30 and 10 cases, respectively.…”

Section: Methodsmentioning

confidence: 99%

Detection of clonal antigen receptor gene rearrangement in dogs with lymphoma by real-time polymerase chain reaction and melting curve analysis

et al. 2014

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BackgroundMolecular techniques that detect canine lymphoma cells by their clonal antigen receptor gene rearrangement play an increasing role for diagnosis as well as for monitoring minimal residual disease during and after cytostatic therapy. However, the methods currently available are time-consuming and/or cost-intensive thus impeding the use in clinical routine. The aim of the present study was to develop and evaluate a real-time polymerase chain reaction (PCR) with subsequent melting curve analysis (MCA) for the detection of clonally rearranged antigen receptor genes in dogs with B and T cell lymphoma on non formalin-fixed and paraffin-embedded lymph node samples.ResultsIn lymph node aspirates from 30 dogs with multicentric B cell lymphoma, real-time PCR with MCA detected clonal rearrangement in 100% and conventional PCR with polyacrylamide gel electrophoresis (PAGE) in 93% of samples. Both methods correctly identified clonality in 80% of lymph node aspirates of 10 dogs with T cell lymphoma. None of the two PCR systems detected clonal rearrangement in samples from 9 dogs with lymph node hyperplasia. Using a dilutional series with regular lymphoid desoxyribonucleic acid (DNA), detection limits of lymphoma DNA were as low as 0.8% and 6.25% for B and T cell clonal rearrangement with real-time PCR and MCA and at 3.13% and 12.5% with the conventional system. Median absolute detection limits of lymphoma DNA were shown to be at 0.1 ng and 1 ng for the B and T cell immunophenotype with the real-time PCR system and at 10 ng each with conventional PCR and PAGE.ConclusionsReal-time PCR with MCA is a convenient and reliable method with a good analytical sensitivity. Thus, the method may assist the detection of clonal antigen receptor gene rearrangement in canine lymphoma patients in a clinical setting also in the presence of small amounts of neoplastic cells.

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Flow cytometric evaluation of peripheral blood and bone marrow and fine-needle aspirate samples from multiple sites in dogs with multicentric lymphoma

Abstract: Results suggested that flow cytometry may aid in detection of extranodal lymphoma involvement in dogs, but further research is needed. Most fine-needle aspirates of liver and spleen were suitable for flow cytometric evaluation.

Cited by 17 publications

References 42 publications

Lymphoma Immunophenotype of Dogs Determined by Immunohistochemistry, Flow Cytometry, and Polymerase Chain Reaction for Antigen Receptor Rearrangements

Lymphoma Immunophenotype of Dogs Determined by Immunohistochemistry, Flow Cytometry, and Polymerase Chain Reaction for Antigen Receptor Rearrangements

Splenic and hepatic ultrasound and cytology in canine lymphoma: effects of findings on stage migration and assessment of prognosis

Detection of clonal antigen receptor gene rearrangement in dogs with lymphoma by real-time polymerase chain reaction and melting curve analysis

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