Validation of human haptoglobin immunoturbidimetric assay for detection of haptoglobin in equine and canine serum and plasma.
Abstract: The Incstar(R) SPQ II human haptoglobin (Hpt) (Incstar Corporation, Stillwater, MN) immunoturbidimetric assay was validated for the determination of serum and plasma Hpt concentrations in dogs and horses. The anti-human Hpt antiserum supplied with the assay, displayed monospecificity to both dog and horse serum Hpt by immunoelectrophoresis and Western blotting techniques. The automated immunoturbidimetric assay results correlated well with the cyanmethemoglobin binding assay (r=0.953 for canine serum and r=0.941 for equine serum), and had excellent precision at both high and low serum Hpt concentrations (within run and between run coefficients of variation near or less than 5%). The assay was linear in both species by serial dilution of pooled-high serum with pooled-low serum, saline and with Hpt-free serum. Interference from hemolysis (> 25 mg/dl hemoglobin) and lipemia greater than 100 mg/dl caused a false decrease and false increase respectively in Hpt yield with the immunoturbidimetric assay. The anti-Hpt antibody supplied with the assay kit, once diluted with polymer diluent and stored at 4 degrees C, was stable for up to 6 days and gave consistent results.
Publication Date: 1996-01-01 PubMed ID: 12660962DOI: 10.1111/j.1939-165x.1996.tb00988.xGoogle Scholar: Lookup
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Summary
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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.
Cite This Article
APA
Weidmeyer CE, Solter PF.
(1996).
Validation of human haptoglobin immunoturbidimetric assay for detection of haptoglobin in equine and canine serum and plasma.
Vet Clin Pathol, 25(4), 141-146.
https://doi.org/10.1111/j.1939-165x.1996.tb00988.x Publication
Researcher Affiliations
- Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Illinois, 2001 South Lincoln Avenue, Urbana, IL 61801.
Citations
This article has been cited 6 times.- Horvatić A, Gelemanović A, Pirkić B, Smolec O, Beer Ljubić B, Rubić I, Eckersall PD, Mrljak V, McLaughlin M, Samardžija M, Lipar M. Multi-Omics Approach to Elucidate Cerebrospinal Fluid Changes in Dogs with Intervertebral Disc Herniation.. Int J Mol Sci 2021 Oct 28;22(21).
- Del Romero A, Cuervo B, Peláez P, Miguel L, Torres M, Yeste M, Rivera Del Alamo MM, Rubio CP, Rubio M. Changes in Acute Phase Proteins in Bitches after Laparoscopic, Midline, and Flank Ovariectomy Using the Same Method for Hemostasis.. Animals (Basel) 2020 Nov 27;10(12).
- Contreras-Aguilar MD, Escribano D, Martínez-Subiela S, Martín-Cuervo M, Lamy E, Tecles F, Cerón JJ. Changes in saliva analytes in equine acute abdominal disease: a sialochemistry approach.. BMC Vet Res 2019 Jun 6;15(1):187.
- Brady N, O'Reilly EL, McComb C, Macrae AI, Eckersall PD. An immunoturbidimetric assay for bovine haptoglobin.. Comp Clin Path 2019;28(1):21-27.
- Freyburger L, Lemaitre L, Médaille C, Oberli F, Fanchon L, Bergamo P. [Comparative safety study of two commercialised vaccines against canine babesiosis induced by Babesia canis].. Parasite 2011 Nov;18(4):311-8.
- Tecles F, Subiela SM, Petrucci G, Panizo CG, Cerón JJ. Validation of a commercially available human immunoturbidimetric assay for haptoglobin determination in canine serum samples.. Vet Res Commun 2007 Jan;31(1):23-36.
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