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Effect of blood sample handling post-collection on Erysipelothrix rhusiopathiae antibody titres
Eric J Neumann1, Kathryn N Bonistalli
1EpiCentre, Institute of Veterinary, Animal and Biomedical Sciences, Massey University, B2.10 Wool Building, Palmerston North, New Zealand. e.neumann@massey.ac.nz
Abstract:
A study was conducted to determine the effect of blood sample mishandling on the performance of an enzyme-linked immunosorbent assay for the detection of antibodies against Erysipelothrix rhusiopathiae. Eleven sample maltreatments (storage at -10 degrees C, storage at 4 degrees C, heat treatment of clotted blood, haemolysis, repetitive freeze-thaw cycling, and substitution of plasma in place of serum) were simulated in a laboratory environment and then run concurrently against a gold standard sample (storage at -80 degrees C). The mishandling treatment groups that simulated high levels of haemolysis had significantly lower optical density (OD) readings when compared to the gold standard. However, the magnitude of the effects was relatively small and only samples with OD values close to the cut-off changed state from positive to negative. Heat treatment had a minor, but non-significant, effect on OD values. Findings from this study suggested that immunoglobulin G antibody was stable in the face of most common sample mishandling events.
Insights
Blood sample mishandling, especially hemolysis, slightly impacts Erysipelothrix rhusiopathiae antibody detection via ELISA. However, immunoglobulin G antibody stability suggests most common sample handling issues do not significantly affect test results.
Area of Science:
- Veterinary Immunology
- Diagnostic Assays
- Microbiology
Background:
- Accurate detection of Erysipelothrix rhusiopathiae antibodies is crucial for animal health management.
- Enzyme-linked immunosorbent assays (ELISA) are widely used for antibody detection.
- Understanding the impact of pre-analytical variables like sample mishandling is vital for reliable diagnostic results.
Purpose of the Study:
- To evaluate the effect of common blood sample mishandling on the performance of an ELISA for detecting antibodies against Erysipelothrix rhusiopathiae.
- To determine the robustness of immunoglobulin G (IgG) antibodies under various simulated adverse storage and processing conditions.
Main Methods:
- Eleven distinct sample maltreatment conditions were simulated, including varied temperatures, heat treatment, hemolysis, freeze-thaw cycles, and plasma substitution.
- Treated samples were analyzed using ELISA and compared against a gold standard sample stored at -80°C.
- Optical density (OD) readings were measured to assess assay performance.
Main Results:
- High levels of hemolysis significantly reduced optical density (OD) readings compared to the gold standard.
- The impact of hemolysis was minor, primarily affecting samples near the positive-negative cut-off.
- Heat treatment showed a small, non-significant effect on OD values.
- Most tested mishandling conditions did not lead to significant changes in antibody detection.
Conclusions:
- Immunoglobulin G (IgG) antibodies against Erysipelothrix rhusiopathiae demonstrate considerable stability under most common sample mishandling scenarios.
- While severe hemolysis can slightly impact ELISA results, the overall assay performance is relatively robust.
- These findings support the reliability of ELISA for Erysipelothrix rhusiopathiae antibody detection even with minor sample handling variations.
