Abstract
Measurement of the acute phase serum protein, haptoglobin (Hp), is performed by biochemical methods based on haemoglobin binding, in many veterinary diagnostic laboratories. During attempts to develop a robust biochemical assay for serum Hp it was discovered that serum albumin interfered with the assay system increasing results by as much as 0.28 mg/ml, which could affect interpretation of results especially in species with low normal Hp concentrations. A reagent cocktail (SB-7) was devised which inhibited the interfering effect of albumin. An automated assay for Hp utilising SB-7 was developed for production as a biochemical assay kit and was evaluated for use in veterinary diagnosis. The intra-assay coefficients of variation were of 0.9%, 0.9% and 1.3% for Hp concentrations of 2.0, 1.0 and 0.23 mg/ml, respectively and interassay coefficients of variation of 1.7% and 4.5% for Hp of 2.08 mg/ml and 0.24 mg/ml, respectively. The lower limit of detection of was 0.02 mg/ml, linearity extended to 8 mg/ml and recovery was 101±7% (mean ±SD). The assay had correlation coefficients (R2) of 0.96 and 0.90 when compared with immunodiffusion assays of canine Hp and bovine Hp, respectively. Lipaemia and bilirubinaemia caused no interference. Haemolysis did not affect measurement of low levels of Hp, but at serum Hp concentrations of 0.4 and 1.8 mg/ml the apparent Hp concentration was decreased. Elevated concentrations of Hp were measured in cattle with mastitis, dogs with polyarthritis and rats experimentally infected withBordetella pertussis. The automated assay is precise and has negligible interference from albumin.
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References
Benesch RE, Kwong S (1990) The stability of the heme-globin linkage in some normal, mutant and chemically modified hemoglobins. J Biol Chem 266:14881–14885
Eckersall PD (1995) Acute phase proteins as markers of inflammatory lesions. Comp Haematol Int 5:93–97
Eckersall PD, Conner JG (1988) Bovine and canine acute phase proteins. Vet Res Comm 12:169–178
Eckersall PD, Parton H, Conner JG et al. (1988) Acute phase reactants in diseases of dog and cattle. In: Blackmore DJ, Eckersall PD, Evans GO et al. (eds) Animal clinical biochemistry: the future. Cambridge University Press, Cambridge, pp 225–230
Eckersall PD, Duthie S, Toussaint MJM et al. (1999) Standardization of diagnostic assays for animal acute phase protein. Ad Vet Med 41:643–655
Eurell TE, Bane DP, Hall WF et al. (1992) Serum haptoglobin concentration as an indicator of weight-gain in pigs. Can J Vet Res 56:6–9
Gattoni M, Boffi A, Sarti P, et al. (1996) Stability of the heme-globin linkage in ab dimers and isolated chains of human hemoglobin. J Biol Chem 271:10130–10136
Hall E, Parton R, Wardlaw AC (1994) Cough production, leucocytosis and serology of rats infected intrabronchially withBordetella pertussis. J Med Microbiol 40:205–213
Harvey JW (1986) Comparison between serum haptoglobin and alpha-2-globulin concentrations in dogs. Vet Clin Pathol 15:4–5
Horadagoda A, Eckersall PD, Hodgson JC et al. (1994) Immediate responses in serum TNFa and acute phase protein concentrations to infection withPasteurella haemolytica A1 in calves. Res Vet Sci 57:129–132
Kushner I, Mackiewicz A (1987) Acute phase proteins as disease markers. Dis Marker 5:1–11
Lipperheide C, Diepers N, Lampreave F et al. (1998) Nephelometric determination of haptoglobin plasma concentrations in fattening pigs. J Vet Med A 45:543–550
Makimura S, Suzuki N (1982) Quantitative determination of bovine serum haptoglobin and its elevation in some inflammatory diseases. Jap J Vet Sci 44:15–21
Schmitt U, Deeg R, Ziegenhorn J (1987) Process and reagents for the determination of the heamoglobin-haptoglobin complex in the presence of free heamoglobin. US Patent no 4,695,552
Skinner JG, Roberts L (1994) Haptoglobin as an indicator of infection in sheep. Vet Rec 134:33–36 (Abstract)
Skinner JG, Brown RAL, Roberts L (1991) Bovine haptoglobin response in clinically defined field conditions. Vet Rec 128:147–149
Solter PF, Hoffmann WE, Hungerford LL et al. (1991) Haptoglobin and ceruloplasmin as determinants of inflammation in dogs. Am J Vet Res 52:1738–1742
Whicher JT, Westacott CI (1992) The acute phase response. In: Whicher JT, Evans SW (eds) Biochemistry of inflammation. Kluwer Academic, London, pp 243–271
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Eckersall, P.D., Duthie, S., Safi, S. et al. An automated biochemical assay for haptoglobin: Prevention of interference from albumin. Comp Haematol Int 9, 117–124 (1999). https://doi.org/10.1007/BF02600369
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DOI: https://doi.org/10.1007/BF02600369