Comparison of microscopic and flow cytometric detection of platelet antibody in dogs suspected of having immune-mediated thrombocytopenia

Annemarie T. Kristensen From the Department of Small Animal Clinical Sciences (Kristensen, Klausner), Department of Veterinary Pathobiology (Weiss), Veterinary Teaching Hospital (Laber), College of Veterinary Medicine, and Department of Laboratory Medicine and Pathology, School of Medicine (Christie), University of Minnesota, St Paul, MN 55108.

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Douglas J. Weiss From the Department of Small Animal Clinical Sciences (Kristensen, Klausner), Department of Veterinary Pathobiology (Weiss), Veterinary Teaching Hospital (Laber), College of Veterinary Medicine, and Department of Laboratory Medicine and Pathology, School of Medicine (Christie), University of Minnesota, St Paul, MN 55108.

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Jeffrey S. Klausner From the Department of Small Animal Clinical Sciences (Kristensen, Klausner), Department of Veterinary Pathobiology (Weiss), Veterinary Teaching Hospital (Laber), College of Veterinary Medicine, and Department of Laboratory Medicine and Pathology, School of Medicine (Christie), University of Minnesota, St Paul, MN 55108.

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Judy Laber From the Department of Small Animal Clinical Sciences (Kristensen, Klausner), Department of Veterinary Pathobiology (Weiss), Veterinary Teaching Hospital (Laber), College of Veterinary Medicine, and Department of Laboratory Medicine and Pathology, School of Medicine (Christie), University of Minnesota, St Paul, MN 55108.

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Douglas J. Christie From the Department of Small Animal Clinical Sciences (Kristensen, Klausner), Department of Veterinary Pathobiology (Weiss), Veterinary Teaching Hospital (Laber), College of Veterinary Medicine, and Department of Laboratory Medicine and Pathology, School of Medicine (Christie), University of Minnesota, St Paul, MN 55108.

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Summary

A flow cytometric platelet immunofluorescence assay (fc-pifa) was compared with a previously developed microscopic platelet immunofluorescence assay (mi-pifa) for detection of circulating platelet antibody. Both assays were performed on serum from 10 healthy dogs with normal platelet count, and on serum from 27 thrombocytopenic dogs—18 had primary immune-mediated thrombocytopenia (imt), and 9 had imt in addition to other immune-mediated disease (secondary imt). Both assays yielded negative results for all control dogs. The mi-pifa and fc-pifa results were in agreement in 23 dogs with imt (14 positive and 9 negative). There was linear correlation between mi-pifa scores and fc-pifa results (r = 0.873). Positive results were obtained for 55.5% of the dogs with suspected imt, using the mi-pifa, compared with 67%, using the fc-pifa; however, the difference was not statistically significant. Use of fresh or frozen fixed donor platelets as the antigen source yielded similar results in the fc-pifa.

Summary

A flow cytometric platelet immunofluorescence assay (fc-pifa) was compared with a previously developed microscopic platelet immunofluorescence assay (mi-pifa) for detection of circulating platelet antibody. Both assays were performed on serum from 10 healthy dogs with normal platelet count, and on serum from 27 thrombocytopenic dogs—18 had primary immune-mediated thrombocytopenia (imt), and 9 had imt in addition to other immune-mediated disease (secondary imt). Both assays yielded negative results for all control dogs. The mi-pifa and fc-pifa results were in agreement in 23 dogs with imt (14 positive and 9 negative). There was linear correlation between mi-pifa scores and fc-pifa results (r = 0.873). Positive results were obtained for 55.5% of the dogs with suspected imt, using the mi-pifa, compared with 67%, using the fc-pifa; however, the difference was not statistically significant. Use of fresh or frozen fixed donor platelets as the antigen source yielded similar results in the fc-pifa.

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