Sandwich enzyme-linked immunosorbent assay for the detection of bovine blood in animal feed.

UNLABELLED The feeding of ruminant proteins to ruminants is prohibited in most countries because the practice is thought to be responsible for the spread of bovine spongiform encephalopathy. However, currently available methods to detect ruminant blood products in rendered feedstuffs are inadequate because they lack species specificity, tissue specificity, and are not based on a thermostable analyte. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for this study that provides reliable and sensitive (0.05-0.5% v/v) detection of bovine blood materials in animal feed. The new sandwich ELISA employs two previously developed monoclonal antibodies (MAbs), Bb6G12 as the capture antibody and biotinylated MAb Bb3D6 as the detecting antibody, and is bovine-specific and blood-specific. The assay is based on the detection of a 60 kDa thermostable protein in bovine blood and provides a useful regulatory tool for monitoring fraudulent labeling or contamination of bovine blood in both heat-processed feedstuffs and unprocessed raw materials. KEYWORDS Bovine; blood; monoclonal antibody; sandwich ELISA.

[1]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[2]  A. Rasooly,et al.  Detection and analysis of animal materials in food and feed. , 2000, Journal of food protection.

[3]  D. Matthews,et al.  The potential for transmissible spongiform encephalopathies in non-ruminant livestock and fish. , 2003, Revue scientifique et technique.

[4]  B. Friguet,et al.  A convenient enzyme-linked immunosorbent assay for testing whether monoclonal antibodies recognize the same antigenic site. Application to hybridomas specific for the beta 2-subunit of Escherichia coli tryptophan synthase. , 1983, Journal of immunological methods.

[5]  W. Liang,et al.  Multiplex polymerase chain reaction method for detection of bovine materials in foodstuffs. , 2003, Journal of AOAC International.

[6]  G. Brambilla,et al.  Trends in animal feed composition and the possible consequences on residue tests , 2005 .

[7]  R. Osta,et al.  Beef- and bovine-derived material identification in processed and unprocessed food and feed by PCR amplification. , 2002, Journal of agricultural and food chemistry.

[8]  Jason R Newgard,et al.  Novel method for detecting bovine immunoglobulin G in dried porcine plasma as an indicator of bovine plasma contamination. , 2002, Journal of agricultural and food chemistry.

[9]  T. Hampton.  New anti-BSE rules imposed. , 2004, JAMA.

[10]  M. Maher,et al.  Real-time polymerase chain reaction detection of bovine DNA in meat and bone meal samples. , 2002, Journal of food protection.

[11]  E. Rencová,et al.  Identification of bovine-specific DNA in feedstuffs. , 2001, Journal of food protection.

[12]  S. V. C. Souza,et al.  In-house validation of a method for detection of animal meals in ruminant feeds by microscopy , 2006 .

[13]  A. Hensel,et al.  Homogeneity of the prion protein gene in various European and Asian pig breeds. , 2004, Journal of veterinary medicine. B, Infectious diseases and veterinary public health.

[14]  J. Ofori,et al.  Monoclonal antibodies specific to thermostable proteins in animal blood. , 2007, Journal of agricultural and food chemistry.