| Type of Document |
Thesis |
| Author |
Ofori, Jack Appiah
|
| Author's Email Address |
papajackx@yahoo.co.uk |
| URN |
etd-07072006-154429 |
| Title |
A sandwich ELISA for detecting bovine blood in ground beef and animal feed |
| Degree |
Master of Science |
| Department |
Nutrition, Food, and Exercise Science, Department of |
| Advisory Committee |
| Advisor Name |
Title |
| Yun-Hwa Peggy Hsieh |
Committee Chair |
| Jodee Dorsey |
Committee Member |
| Mary Ann Moore |
Committee Member |
| Shridhar K. Sathe |
Committee Member |
|
| Keywords |
- Blood Infectivity
- Transmissible Spongiform Encephalopathies
- Prions In Blood
|
| Date of Defense |
2006-06-30 |
| Availability |
unrestricted |
Abstract
Bovine plasma proteins are used as high quality ingredients in feed for farm animals and also as a binder or colorant in ground beef. Religious observance, as well as recent fears of epidemic bovine spongiform encephalopathy, calls for suitable methods to detect bovine blood in processed food and animal feed for regulatory purposes. Analytical methods are currently not available for this purpose because the available methods are neither bovine specific, tissue specific nor based on the detection of a thermal-stable analyte in blood. This research sought to develop a sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of bovine blood in human food and animal feed. Two bovine-specific monoclonal antibodies (Mab 6G12 and Mab 3D6) that bind to the same antigenic thermal-stable protein (60kDa) in heat-treated bovine blood were selected to construct the sandwich ELISA. Mab 6G12 was used as the coating antibody and biotinylated Mab 3D6 was used as the detecting antibody. Soluble proteins were extracted with 10 mM phosphate buffered saline (PBS, pH 7.2). This sandwich ELISA was bovine specific, with no cross reactivity to other bovine tissues or other common food additive proteins such as soy protein, egg albumin, and gelatin. The assay, however, showed trace cross-reaction with non-fat dry milk. The cross-reactivity can be eliminated when antibody dilution is increased. The assay had a detection limit of 0.1% of spray-dried bovine plasma in spray-dried porcine plasma and 0.5% whole bovine blood powder in spray- dried porcine plasma, with a sensitivity of 100%, specificity of 100% and an overall accuracy of 100%. Low levels (1% v/w) of bovine blood in raw and cooked beef were successfully detected by the sandwich assay. The assay is suitable for detecting bovine blood in animal feed as well as cooked and fresh ground beef because it is bovine specific, tissue specific and based on a thermo-stable bio-marker.
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