Validation of human haptoglobin immunoturbidimetric assay for detection of haptoglobin in equine and canine serum and plasma.

The research validates the use of a human haptoglobin immunoturbidimetric assay for detecting haptoglobin levels in equine and canine blood. The results show a high correlation with other testing methods and noted precision across different concentrations, but revealed potential for interference from hemolysis and high lipid levels.

Validation of Human Haptoglobin Immunoturbidimetric Assay for Equine and Canine Use

In this study, an immunoturbidimetric assay (a laboratory test commonly used to determine the concentrations of proteins in a liquid) originally designed for human use was tested for its effectiveness in determining serum and plasma haptoglobin concentrations in dogs and horses:

• The Incstar(R) SPQ II human haptoglobin (Hpt) was the evaluated immunoturbidimetric assay.
• Researchers found that the anti-human Hpt antiserum included in the assay kit demonstrated a monospecific response, indicating that it reacts specifically with the Hpt in both dog and horse serum.
• This was confirmed through immunoelectrophoresis and Western blotting, laboratory techniques used to separate proteins by size and charge.

Performance of the Assay

The efficacy of the immunoturbidimetric assay was compared to that of the cyanmethemoglobin binding assay:

• The results from the two tests correlated well, indicating that the immunoturbidimetric assay is a reliable method for measuring haptoglobin concentrations in canine and equine serum.
• The precision of the test was also noteworthy, with both high and low serum Hpt concentrations showing consistent results with minimal variation.

Interference and Limitations of the Assay

The study also identified potential limitations of the immunoturbidimetric assay:

• Hemolysis (the breakdown of red blood cells) and high lipid levels were found to cause false results, either an artificial decrease or an increase in Hpt yield respectively.
• The amount of interference observed was proportional to the degree of hemolysis or lipemia in the serum samples.

Stability of the Anti-Hpt Antibody

The long-term performance of the reagents used in the assay were also evaluated:

• The anti-Hpt antibody supplied with the kit remained stable and gave consistent results when it was diluted with a polymer diluent and stored at a low temperature (4 degrees Celsius) for up to six days.