Immune Netw.  2017 Aug;17(4):269-274. 10.4110/in.2017.17.4.269.

Diagnosis of Lymphoid Malignancy by PCR for Analysis of Antigen Receptor Rearrangement after Blood Transfusion in a Dog with Acute Lymphocytic Leukemia

• Kim S
• Kim H
• Lee SH
• Cho I
• Kang S
• Bae J
• Kim W
• Ahn S
• Choi J
• Kim SK
• Do Y
• Yoo JG
• Park J
• Yu D

• KMID: 2388081
• DOI: http://doi.org/10.4110/in.2017.17.4.269

Abstract

Acute lymphocytic leukemia (ALL) is uncommon lymphoid malignancy in dogs, and its diagnosis is challenging. A 14-year-old spayed female mixed breed dog was transferred to a veterinary medical teaching hospital for an immediate blood transfusion. The dog showed lethargy, pale mucous membranes, and a weak femoral pulse. Complete blood count revealed non-regenerative anemia and severe leukopenia with thrombocytopenia. ALL was tentatively diagnosed based on the predominance of immature lymphoblasts on blood film examination. For confirmation of lymphoid malignancy, PCR for antigen receptor rearrangement (PARR) on a peripheral blood sample and flow cytometry analysis were performed after blood transfusion. Flow cytometry analysis revealed that lymphocyte subsets were of normal composition, but PARR detected a T-cell malignancy. The dog was diagnosed with ALL and survived 1 wk after diagnosis. In conclusion, after blood transfusion, flow cytometry was not a reliable diagnostic method for an ALL dog, whereas PARR could detect lymphoid malignancy. Our results suggest that PARR should be the first-line diagnostic tool to detect canine lymphoid malignancy after a blood transfusion.

Keyword

Acute lymphoblastic leukemia; PARR; Flow cytometry; Blood transfusion; Lymphoid malignancy

MeSH Terms

Adolescent
Anemia
Animals
Blood Cell Count
Blood Transfusion*
Diagnosis*
Dogs*
Female
Flow Cytometry
Hospitals, Teaching
Humans
Lethargy
Leukopenia
Lymphocyte Subsets
Methods
Mucous Membrane
Polymerase Chain Reaction*
Precursor Cell Lymphoblastic Leukemia-Lymphoma*
Receptors, Antigen*
T-Lymphocytes
Thrombocytopenia
Receptors, Antigen

Figure

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  Figure 1 (A) Microscopic examination of peripheral blood prior to administration. (B) Microscopic examination of feathered edge of blood film on admission. (C) PCR for analysis of antigen receptor rearrangement from peripheral blood. First lane ‘M’ shows a 25 bp size marker, lane ‘C’ indicates positive control for DNA, lane 3 and 4 detect “major” and “minor” immunoglobulin rearrangements, respectively, and lane 5 (TCR) detects TCRγ gene rearrangement. (D) A dot plot from flow cytometry analysis. Immature lymphoid cells were prominent prior to administration but microscopic examination was not easy because of severe leukopenia on admission. PARR revealed positive for clonal rearrangement of TCRγ gene but flow cytometry did not show overt T cell predominance after blood transfusion in this dog. TCR, T-cell receptor; PARR, PCR for antigen receptor rearrangement.

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