Sandwich enzyme-linked immunosorbent assay for quantitative measurement of serum amyloid A protein in horses

Megumi Satoh From the Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060, Japan.

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Toru Fujinaga From the Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060, Japan.

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Masahiro Okumura From the Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060, Japan.

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Mitsuyoshi Hagio From the Laboratory of Veterinary Surgery, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060, Japan.

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SUMMARY

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 10 to 30 μg/ml. Mean (± sd) concentrations of saa in foals (≤ 12 months old) and adult horses (≥ 18 months old) were 21.23 ± 12.20 and 14.93 ± 9.07 μg/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 μg/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.

SUMMARY

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 10 to 30 μg/ml. Mean (± sd) concentrations of saa in foals (≤ 12 months old) and adult horses (≥ 18 months old) were 21.23 ± 12.20 and 14.93 ± 9.07 μg/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 μg/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.

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