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Sandwich enzyme-linked immunosorbent assay for quantitative measurement of serum amyloid A protein in horses.
Abstract: To measure the concentration of serum amyloid A (sAA) protein in horses, a sensitive and highly reproducible sandwich (ELISA) was established, using affinity purified SAA antibody. Results of the ELISA were found to have a high correlation (r = 0.95) with those of the single radial immunodiffusion test. Equine SAA concentration was measured by use of this ELISA. In clinically normal horses, the concentration of SAA was high immediately after birth to 2 weeks of age. After that, SAA concentration had periodic fluctuations in the range of approximately 1.0 to 30 micrograms/ml. Mean (+/- SD)) concentrations of SAA in foals (< or = 12 months old) and adult horses (> or = 18 months old) were 21.23 +/- 12.20 and 14.93 +/- 9.07 micrograms/ml, respectively. In mares during the perinatal period, SAA concentration remained stable within the reference range before parturition. It increased quickly after delivery, and reached a peak value of 101.29 +/- 98.82 micrograms/ml on postpartum day 3, then began to decrease, at postpartum week 2, to the reference range by the end of postpartum month 1. In horses with experimentally induced inflammation, SAA concentration increased quickly and reached approximately four- to 40-fold increase over the pretreatment value on day 1 and remained high on days 2 to 6 after treatment. It then returned to the baseline value by 2 to 4 weeks in association with disappearance of local signs of inflammation. The SAA concentration was high in most horses with clinical signs of inflammation. It was concluded from these data that this ELISA is sensitive and reliable for measuring SAA in horses.
Publication Date: 1995-10-01 PubMed ID: 8928944 The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research involves the use of a sandwich enzyme-linked immunosorbent assay (ELISA) for quantifying the serum amyloid A (SAA) protein concentrations in horses. The results demonstrate that the technique is highly sensitive and reliable.
Research Methodology
- The researchers developed a sandwich ELISA, utilizing an affinity-purified SAA antibody, to measure SAA protein concentration in horses.
- Once the ELISA was established, the researchers tested its accuracy by comparing the results with a frequently used other method, the single radial immunodiffusion test. They found a strong correlation between the two approaches, with a correlation coefficient (r) of 0.95.
Equine SAA Concentration Measurements
- With the new ELISA, the team examined the SAA concentration in healthy horses and discovered that it was high immediately after birth until the horse was two weeks old. Following this period, the concentration fluctuated between 1.0 to 30 micrograms/ml.
- The researchers also looked at the differences between different horse age groups. The mean concentration of SAA in foals, 12 months or younger, was higher (21.23 +/- 12.20) micrograms/ml than the concentration in adults, 18 months or older, (14.93 +/- 9.07) micrograms/ml.
SAA concentration in Mares and Horses with Induced-Inflammation
- For mares in the perinatal period, the SAA concentration stayed within the reference range before parturition, increased shortly after giving birth, peaked on the third postpartum day, and then decreased to the reference range by the end of the first postpartum month.
- In horses where inflammation was experimentally induced, the SAA concentration increased rapidly reaching four to forty times the pre-treatment value on the first day, remaining high for up to six days following the treatment. The concentration then returned to baseline between two to four weeks into the treatment correlating with the disappearance of inflammation symptoms.
- The level of SAA was elevated in most horses that exhibited inflammation signs.
Research Conclusion
Finally, the team concluded that their ELISA technique for measuring SAA in horses was both sensitive and reliable. The findings suggest a significant relationship between SAA concentrations and inflammation, and the potential for monitoring the protein as an indicator of inflammation in equines.
Cite This Article
APA
Satoh M, Fujinaga T, Okumura M, Hagio M.
(1995).
Sandwich enzyme-linked immunosorbent assay for quantitative measurement of serum amyloid A protein in horses.
Am J Vet Res, 56(10), 1286-1291.
Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
MeSH Terms
- Amino Acids / analysis
- Animals
- Animals, Newborn / blood
- Castration / veterinary
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Horse Diseases / blood
- Horses / blood
- Immunodiffusion / veterinary
- Inflammation / blood
- Inflammation / veterinary
- Jejunostomy / veterinary
- Male
- Pregnancy
- Rabbits
- Reference Values
- Reproducibility of Results
- Sensitivity and Specificity
- Serum Amyloid A Protein / analysis
- Serum Amyloid A Protein / chemistry
- Serum Amyloid A Protein / immunology
Citations
This article has been cited 7 times.- Souto PC, Santos MR, Orozco AMO, Bento LD, Ramirez-Lopez CJ, Girardi FM, Machado JCA, de Oliveira LL, da Fonseca LA. Enzyme-Linked Immunosorbent Assay (ELISA) Development for Equine Serum Amyloid A (SAA) Determination Using Recombinant Proteins. Methods Protoc 2025 Apr 7;8(2).
- Satué K, Calvo A, Muñoz A, Fazio E, Medica P. Interrelationship between reproductive hormones and acute phase proteins during estrous cycle and pregnancy in Spanish purebred broodmares. Vet Anim Sci 2021 Dec;14:100212.
- Valigura HC, Leatherwood JL, Martinez RE, Norton SA, White-Springer SH. Dietary supplementation of a Saccharomyces cerevisiae fermentation product attenuates exercise-induced stress markers in young horses. J Anim Sci 2021 Aug 1;99(8).
- Robinson CS, Singer ER, Piviani M, Rubio-Martinez LM. Are serum amyloid A or D-lactate useful to diagnose synovial contamination or sepsis in horses?. Vet Rec 2017 Oct 21;181(16):425.
- Pihl TH, Scheepers E, Sanz M, Goddard A, Page P, Toft N, Andersen PH, Jacobsen S. Influence of disease process and duration on acute phase proteins in serum and peritoneal fluid of horses with colic. J Vet Intern Med 2015 Mar-Apr;29(2):651-8.
- Hillström A, Tvedten H, Lilliehöök I. Evaluation of an in-clinic Serum Amyloid A (SAA) assay and assessment of the effects of storage on SAA samples. Acta Vet Scand 2010 Feb 2;52(1):8.
- Hultén C, Sandgren B, Skiöldebrand E, Klingeborn B, Marhaug G, Forsberg M. The acute phase protein serum amyloid A (SAA) as an inflammatory marker in equine influenza virus infection. Acta Vet Scand 1999;40(4):323-33.
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