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Detection of staphylococcal enterotoxin B by affinity radioimmunoassay

Journal Article · · J. Food Sci.; (United States)

The objective of this investigation was to develop a rapid, sensitive method for the detection and quantitation of staphylococcal enterotoxin B (SEB). Antibody to SEB was covalently attached to Sepharose 4B by use of cyanogen bromide activation. The antibody gel was diluted with Sephadex G-25, and 1 to 2 ml packed volume of the diluted gel was placed in a small column. Assays were carried out by sequentially pumping sample, /sup 125/I-labeled SEB, 0.05M Na-phosphate buffer (pH 7.5) containing 0.15M NaCl, and 0.13M NH/sub 4/OH (pH 10.5) containing 0.15M NaCl through the column. The amount of /sup 125/I-labeled SEB removed from the column with the pH 10.5 solution was inversely related to the amount of SEB in the sample. After washing with the pH 7.5 buffer, the column was ready to reuse. Columns were used for over 100 determinations. The minimum SEB concentrations which could be detected were 1.2 ng/ml in buffer, 2.2 ng/ml in nonfat dry milk, and 6.3 ng/g in hamburger. This affinity radioimmunoassay (RIA) was also suitable for measuring thermal inactivation of SEB. Results of this investigation show that this affinity RIA technique can be utilized for rapid detection of SEB.

Research Organization:
Michigan State Univ., East Lansing
OSTI ID:
7075862
Journal Information:
J. Food Sci.; (United States), Journal Name: J. Food Sci.; (United States) Vol. 43:3; ISSN JFDSA
Country of Publication:
United States
Language:
English