Additional file 1 of Comparative study of immunohematological tests with canine blood samples submitted for a direct antiglobulin (Coombs’) test

Additional file 1: Supplementary Table S1. Test conditions for six direct antiglobulin test (DAT) methods. Supplementary Table S2. Demographic data of 126 dogs suspected to have immune-mediated hemolytic anemia (IMHA) and 28 healthy control dogs and with DAT+ results by microtiter. Plate direct antiglobulin test (DAT) with using goat anti-dog IgG, IgM, and C3 at 22°C. Breed (A), age (B), and gender (C) distribution. Supplementary Tables S3. Demographic data of 126 dogs suspected to have immune-mediated hemolytic anemia (IMHA) with positive and negative direct antiglobulin test (DAT) results. Breed (A), age (B), and gender (C) distribution. Supplementary Table S4.In vitro-induced positive direct antiglobulin test (DAT) results in samples from eight healthy dogs with DEA 4+, DEA 5- and Dal+ blood type by adding anti-DEA 4, anti-DEA 5, and anti-Dal antisera. Supplementary Table S5. Anemia, reticulocytosis, hyperbilirubinemia, and hemolyzed plasma compared to DAT results in dogs suspected to have IMHA. Supplementary Table S6. Degree of anemia (total hemoglobin [Hb], packed cell volume [PCV], and hematocrit [Hct]) in relation to direct antiglobulin test (DAT) results in dogs suspected to have immune-mediated hemolytic anemia. Supplementary Table S7. Correlation between polychromasia and reticulocytosis of samples from 80 dogs suspected to have immune-mediated hemolytic anemia and MICRO GAD direct antiglobulin test. Results from DAT+ dogs in brackets. Supplementary Table S8. Microtiter plate direct antiglobulin test (DAT) results with two antiglobulins (goat anti-dog; GAD and rabbit anti-dog; RAD) and at three different temperatures (22, 37 and 4°C) for 126 dogs suspected to have immune-mediated hemolytic anemia. Supplementary Table S9. Comparison of six different direct antiglobulin tests (DAT) with one to two different antiglobulin reagent. Supplementary Table S10. Minitube gel kit direct antiglobulin test (GEL KIT DAT) in dogs suspected to have immune-mediated hemolytic anemia with and without prior 3x washing of red blood cells. Supplementary Table S11. Comparison of six direct antiglobulin test (DAT) results in 126 dogs suspected to have immune-mediated hemolytic anemia (IMHA). The Cohen’s kappa (κ) values and a two-sided confidence intervals (95%) is reported for each pair of methods used. Supplementary Table S12. Comparison of six direct antiglobulin test (DAT) results in 126 dogs suspected to have immune-mediated hemolytic anemia (IMHA) and 28 healthy dogs. The Cohen’s kappa (κ) values and two-sided confidence intervals (95%) are reported for each pair of methods used. Supplement Table S13. Cohen’s kappa (κ) value of the spherocytosis, and macroscopic in tube agglutination and macroscopic saline agglutination test (SAT) results at two dilutions compared to the microtiter plate DAT method with goat-anti dog IgG, IgM, and C3 at 22°C. Supplementary Figure S1. Flow chart oflaboratory tests and six different direct antiglobulin test (DAT) methods performed with blood samples from dogs suspected to have immune-mediated hemolytic anemia. Supplementary Figure S2. Severity of anemia, reticulocytosis, hyperbilirubinemia, and free plasma hemoglobin concentration compared to microtiter plate direct antiglobulin test (MICRO DAT) results in dogs suspected to have immune-mediated hemolytic anemia. Supplementary Figure S3. Severity of anemia, reticulocytosis, bilirubinemia, and free plasma hemoglobin concentration results grouped based on ≥4 DAT+ results in dogs suspected to have immune-mediated hemolytic anemia