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Direct
immunofluorescent assay for isotype specific antibodies coating
erythrocytes in dogs and horses with immune mediated anemia
The Direct Immunofluorescent assay detects antibodies on red blood cells.
For dogs, we use fluorescent labeled polyclonal antisera to canine IgG,
IgM, IgA, and C3. For horses, we use fluorescent labeled polyclonal
antisera to equine IgG, IgM and IgA. This test is more sensitive that the
Coomb’s anti-globulin agglutination test, because it is not affected by
the prozone effect. The prozone effect is when the agglutination based
Coomb’s test is negative at low dilutions of Coomb’s reagent. The
sensitivity is 100% and the specificity of the direct immunofluorescent
assay is 87.5% determined by population of dogs with immune mediated
hemolytic anemia (J. Vet. Intern. Med. 2000, 14:190-196). The test does
not differentiate primary (idiopathic or autoimmune) immune mediated
hemolytic anemia (IMHA) from secondary causes for IMHA. We have identified
antibodies on the surface of equine red cells in cases of penicillin
induced IMHA, clostridial myositis, and neonatal isoerythrolysis.
Direct
Immunofluorescent Assay
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