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Detection of cows' milk in ewes' milk and cheese by an indirect enzyme-linked immunosorbent assay (ELISA)

Published online by Cambridge University Press:  01 June 2009

Elena Rodríguez ,
Rosario Martín ,
Teresa García ,
Pablo E. Hernández  and
Bernabé Sanz
Elena Rodríguez
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Rosario Martín
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Teresa García
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Pablo E. Hernández
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
Bernabé Sanz
Affiliation:
Departamento de Higiene y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain
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Summary

An indirect enzyme-linked immunosorbent assay was successfully developed for the detection of defined amounts of cows' milk (1–50%) in sheeps' milk and cheese. The assay used polyclonal antibodies raised in rabbits against bovine caseins (BC). The anti-BC antibodies were recovered from the crude antiserum by immunoadsorption and elution from a column containing BC. The antibodies were biotinylated and rendered cows' milk specific by mixing them with lyophilized ovine and caprine caseins. ExtrAvidin-peroxidase was used to detect the biotinylated anti-BC antibodies bound to BC immobilized on 96-well plates. Subsequent enzymic conversion of substrate gave clear absorbance differences when assaying mixtures of sheeps' milk and cheese containing variable amounts of cows' milk.

Type
Original Articles
Information
Journal of Dairy Research , Volume 57 , Issue 2 , May 1990 , pp. 197 - 205
Copyright
Copyright © Proprietors of Journal of Dairy Research 1990

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References

REFERENCES

Addeo, F., Anelli, G., Stingo, C., Chianese, L., Petrilli, P. & Scudiero, A. 1984 [Detection and determination of bovine milk in Pecorino cheese.] Latte 9 37–44Google Scholar
Albonico, F. & Resmini, P. 1967 [Determination of caseins in Mozzarella cheese by cellulose acetate electrophoresis.] Bolletino del Laboratorio Chimico Provinciale 18 143–152Google Scholar
Aranda, P., Oria, R. & Calvo, M. 1988 Detection of cows' milk in ewes' milk and cheese by an immunodotting method. Journal of Dairy Research 55 121–124Google Scholar
Aschaffenburg, R. & Dance, J. E. 1968 Detection of cows' milk in goats' milk by gel electrophoresis. Journal of Dairy Research 35 383–384Google Scholar
Assenat, L. 1967 [A method of identifying milks and cheeses by means of polyacrylamide-gel electrophoresis.] Lait 47 393–414, 496–502CrossRefGoogle Scholar
Benassi, R. 1963 [The compositions of fat of ewes' milk.] Latte 37 468–470Google Scholar
Bican, P. 1983 High performance liquid chromatography of casein peptides. Journal of Dairy Science 66, 2195–2197Google Scholar
Bican, P. & Blanc, B. 1982 Milk protein analysis: a high-performance chromatography study. Milchwissenschaft 37 592–593Google Scholar
Cerbulis, J., Parks, O. W. & Farrell, H. M., Jr. 1982 Composition and distribution of lipids of goats' milk. Journal of Dairy Science 65 2301–2307CrossRefGoogle Scholar
Diosady, L. L., Bergen, I. & Harwalkar, V. R. 1980 High performance liquid chromatography of whey proteins. Milchwissenschaft 35 671–674.Google Scholar
Doberstein, K. H. & Greuel, E. 1982 [Preparation of specific antisera for identification of meat of African antelope species in an agar gel precipitation test.] Fleischwirtscaft 62 1011–1013Google Scholar
Durand, M., Meusnier, M., Delahaye, J. & Prunet, P. 1974 [Detection of cows' milk in goats' and ewes' milk by an immunodiffusion method.] Bulletin de l'Académie Vétérinaire de France 47 247–258Google Scholar
Furtado, M. M. 1983 Detection of cow milk in goat milk by polyacrylamide gel electrophoresis. Journal of Dairy Science 66 1822–1824Google Scholar
García, T., Martin, R., Rodriguez, E., Hernández, P. E. & Sanz, B. 1989 Development of a cows' milk identification test (COMIT) for field use. Journal of Dairy Research 56 691–698Google Scholar
Hillier, R. M. 1976 The quantitative measurement of whey proteins using polyacrylamide-gel electrophoresis. Journal of Dairy Research 43 259–265CrossRefGoogle ScholarPubMed
Iverson, J. L. & Sheppard, A. J. 1985 Utilization of capric and lauric acid ratios to detect substitution of cow's milk in imported sheep and goat cheeses. Journal of Dairy Science 68 (Suppl. 1) 95Google Scholar
Jones, S. J. & Patterson, R. L. S. 1986 A modified indirect ELISA procedure for raw meat speciation using crude anti-species antisera and stabilised immunoreagents. Journal of the Science of Food and Agriculture 37 767–775.Google Scholar
Kaiser, K. P. & Krause, I. 1985 [Analysis of proteins in foods by electrophoretic and Chromatographie methods.] Zeitschrift für Lebensmittel-Untersuchung und -Forschung 180 181–201Google Scholar
Levieux, D. 1977 [New technique for detecting adulteration of goats' and ewes' milk.] Dossiers de l'Elevage. 2 (2) 37–46Google Scholar
Levieux, D. 1980 The development of a rapid and sensitive method, based on hemagglutination inhibition, for the measurement of cow milk in goat milk. Annales de Recherches Vétérinaires 11 151–156Google Scholar
Lotito, A. & Cucurachi, A. 1967 [Gas Chromatographie examination of ewes' milk fat.] Rivista Italiana della Sostanze Grasse 44 341–348Google Scholar
Ng-Kwai-Hang, K. F. & Kroeker, E. M. 1984 Rapid separation and quantification of major caseins and whey proteins of bovine milk by polyacrylamide gel electrophoresis. Journal of Dairy Science 67 3052–3056Google Scholar
Ouchterlony, O. 1948 In vitro method for testing the toxin producing capacity of diphtheria bacteria. Acta Pathologica et Microbiologica Scandinavica 25 186–191Google Scholar
Palo, V. 1975 Falsification prove of sheep's cheese by cow's cheese. International Dairy Federation, A7-DOC 3Google Scholar
Pearce, R. J. 1983 Analysis of whey proteins by high performance liquid chromatography. Australian Journal of Dairy Technology 38 114–117Google Scholar
Pierre, A. & Portmann, N. A. 1970 [Detection of cows' milk added to goats' milk using polyacrylamide-gel electrophoresis. Application to cheese.] Annales de Technologie Agricole 19 107–130Google Scholar
Radford, D. V., Tchan, Y. T. & McPhillips, J. 1981 Detection of cow's milk in goat's milk by immunoelectrophoresis. Australian Journal of Dairy Technology 36 144–146Google Scholar
Ramos, M. & Juarez, M. 1981 [Analytical criteria for detection of cows' milk in mixtures with milk of other species.] Revista Española de Lecheria No. 121 193–206Google Scholar
Ramos, M. & Juarez, M. 1984 Update on existing analytical methods for detecting mixtures of cow's, ewe's and goat's milk. International Dairy Federation Bulletin Document no. 181Google Scholar
Ramos, M., Martinez-Castro, I. & Juarez, M. 1977 Detection of cow's milk in Manchego cheese. Journal of Dairy Science 60 870–877Google Scholar
Ramos, M., Martinez-Castro, I. & Juarez, M. 1980 [Detection of cows' milk in goats' milk. Application to fresh cheeses.] III Congreso Nacional de Química, Química Agricola Alimentaria, Sevilla 121–127Google Scholar
Ruiz, E. & Santillana, I. 1986 [Determination of mixtures of cows', ewes' and goats' milk by isoelectrofocusing in polyacrylamide gel.] Alimentaria 171 55–64Google Scholar
Sadini, V. 1963 [Fat of ewes' milk and of Italian ewes' milk cheese.] Latte 37 933–942.Google Scholar
Tijssen, P. 1985 Preparation of avidin and biotinylated immunoassay reactants. In Practice and Theory of Enzyme I mmunoassays pp. 23–31Amsterdam: Elsevier Science Publishers (Laboratory Techniques in Biochemistry and Molecular Biology vol. 15)Google Scholar
Tweeten, T. N. & Euston, C. B. 1980 Application of high performance liquid chromatography in the food industry. Food Technology 34 (12) 29–32, 34, 36–37Google Scholar
Wilchek, M. & Bayer, E. A. 1988 The avidin-biotin complex in bioanalytical applications. Analytical Biochemistry 171 1–32Google Scholar