Serum obtained from C57BL/6 mice infected with T. gondii tachyzoites was assayed by enzyme-linked immunosorbent assay (ELISA) using antibodies against the T. gondii P30 protein with a Multispecies ID Screen® Toxoplasmosis Indirect kit (IDVET, Montpellier, France) following the manufacturer’s instructions. The serum was diluted 1:9 with dilution buffer and incubated for 1 h at room temperature. The plates were rinsed three times with washing buffer, after which horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG antibody (Sigma, St. Louis, MO, USA) was added, and then, the plates were incubated for 1 h at room temperature. After the final washing, substrate solution was added to each well and incubated for 15 min at room temperature in the dark. The reaction was stopped using 0.5 M H2SO4 solution, and the absorbance was read at 450 nm using an ELISA reader. Mean OD values > 0.5 were defined as positive [28 (link)].
Multispecies id screen toxoplasmosis indirect kit
Manufactured by IDvet
Sourced in France
The Multispecies ID Screen® Toxoplasmosis Indirect kit is an indirect enzyme-linked immunosorbent assay (ELISA) designed to detect antibodies to Toxoplasma gondii in various animal species. It is used as a diagnostic tool to identify exposure to the Toxoplasma parasite.
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3 protocols using multispecies id screen toxoplasmosis indirect kit
1
ELISA-based Toxoplasmosis Serology
2
Serological Testing for Toxoplasmosis and Neosporosis
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The serum sample was analysed for the presence of antibodies to T. gondii using a multi-species ID Screen Toxoplasmosis Indirect kit (IDvet, Montpellier) and for N. caninum using an ELISA kit (IDEXX Laboratories Inc., Westbrook, ME, USA). The analysis was performed according to the manufacturer’s instructions.
3
Seroprevalence of Toxoplasma gondii in Grey Squirrels
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We collected a minimum of 500 µL of blood from 15 individual grey squirrels and stored samples on ice. All samples were processed within 12 hours. We centrifuged the collected blood samples at 3500 rpm for 15 minutes and froze the serum at -20 °C until used for testing.
Enzyme-linked immunosorbent assay.-We used enzyme-linked immunosorbent assays (ELISA) to detect serum antibodies (IgG) against T. gondii. As species-specific conjugates are not available for squirrels, we used a commercially available ELISA kit for testing the samples (Multi-species ID Screen Toxoplasmosis Indirect kit, IDVet, Grabels, France) following the manufacturer's instructions. We read the optical density values at 450 nm in a spectrophotometer and calculated results using these values and kit controls expressed as S/P (Sample to Positive Ratio) percentage (S/P%). We considered samples with S/P% less or equal to 40% negative; samples with S/P% between 40 and 50% doubtful or inconclusive; and samples with an S/P% higher than 50% positive, following the kit's protocol.
Enzyme-linked immunosorbent assay.-We used enzyme-linked immunosorbent assays (ELISA) to detect serum antibodies (IgG) against T. gondii. As species-specific conjugates are not available for squirrels, we used a commercially available ELISA kit for testing the samples (Multi-species ID Screen Toxoplasmosis Indirect kit, IDVet, Grabels, France) following the manufacturer's instructions. We read the optical density values at 450 nm in a spectrophotometer and calculated results using these values and kit controls expressed as S/P (Sample to Positive Ratio) percentage (S/P%). We considered samples with S/P% less or equal to 40% negative; samples with S/P% between 40 and 50% doubtful or inconclusive; and samples with an S/P% higher than 50% positive, following the kit's protocol.
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