title: Development of an enzyme immuno assay for the determination of porcine haptoglobin in various body fluids: testing the significance of meat juice measurements for quality monitoring programs.
abstract: Quantification of haptoglobin (Hp), an acute phase protein, in blood is presently discussed as being useful to monitor animal health. We developed an enzyme immuno assay (EIA) which is specific for porcine Hp, is not impaired by hemolytic samples and is sufficiently sensitive to be applied in meat juice. Hp was purified from porcine serum by affinity chromatography on hemoglobin Sepharose followed by gel filtration. A specific rabbit antiserum was obtained. In a competitive approach, biotinylated porcine Hp was used as tracer and incubated with Hp standard or sample in microtiter plates. The limit of detection was 0.02 mg/l, parallelism of sample dilutions was proven; recovery of Hp added to serum samples was 96.4 +/- 4.7%. The coefficients of intra and inter-assay variation were 3.3 (n=5) and 10.2% (n=16), respectively. Hp was reliably quantified in blood serum and plasma, whole blood, saliva and meat juice. For healthy pigs of different ages (4 weeks and 6 months), mean Hp concentrations of about 0.5-0.7 mg/ml were observed. To test the significance of Hp measurements in other matrices, samples were obtained from fattening pigs or from slaughter pigs. Blood serum or plasma was collected in parallel. In whole blood, Hp concentrations were about 40% lower than in plasma, but were closely related (n=24,r=0.85,P<0.001). Saliva Hp concentrations ranged between 0.3 and 3.0 microg/ml and were marginally related with blood plasma concentrations (n=93,r=0.35,P<0.001). From 106 hybrid slaughter pigs (100-110 kg) blood and muscle samples (diaphragmatic pillar, d.p.; m. brachiocephalicus, m.b.) were collected. Meat juice was obtained after freezing and thawing. Concentrations were 0.39+/-0.5 mg/ml in serum and 0.04+/-0.06 mg/ml in meat juice. Hp concentrations in blood were closely correlated with those in d.p. juice (P<0.001,r=0.750) and m.b. juice (P<0.001,r=0.776). In view of the many reports on Hp measurements being predictive for animal health even in the subclinical range, we conclude that Hp quantification in meat juice might be useful to assess meat quality at slaughter and further along the processing chain in terms of animal health.
authors.affiliation: Institute for Physiology, Biochemistry and Animal Hygiene, University of Bonn, Katzenburgweg 7-9, D-53115 Bonn, Germany. s.hiss@uni-bonn.de
authors.forename: Stephanie
authors.initials: S
authors.lastname: Hiss
authors.forename: Susanne
authors.initials: S
authors.lastname: Knura-Deszczka
authors.forename: Gertraud
authors.initials: G
authors.lastname: Regula
authors.forename: Mark
authors.initials: M
authors.lastname: Hennies
authors.forename: Stefanie
authors.initials: S
authors.lastname: Gymnich
authors.forename: Brigitte
authors.initials: B
authors.lastname: Petersen
authors.forename: Helga
authors.initials: H
authors.lastname: Sauerwein
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002846:Chromatography, Affinity / Q000662:veterinary; D002850:Chromatography, Gel / Q000662:veterinary; D006242:Haptoglobins / Q000302:isolation & purification*; D007124:Immunoenzyme Techniques / Q000379:methods / Q000662:veterinary*; D008460:Meat / Q000032:analysis* / Q000592:standards; D012463:Saliva / Q000737:chemistry; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000276:immunology
chemical_list: D006242:Haptoglobins