Genigeorgis C, Kuo J K
Appl Environ Microbiol. 1976 Feb;31(2):274-9. doi: 10.1128/aem.31.2.274-279.1976.
Extraction, concentration, and serological detection of staphylococcal enterotoxins from foods are laborious and time consuming. By exposing food extracts to an insoluble matrix tagged with specific anti-enterotoxin B, we have been able to recover the toxin from foods in a sensitive and rapid way. After mixing the reagents for 2 h at room temperature, immunoglobulin G antibodies were attached to CNBr-activated Sepharose 4B at pH 8.5 (0.1 M carbonate buffer with 0.5 M NaCl). Sepharose-antibody complex (1 ml) specifically recovered 0.1 to 30 mug of enterotoxin B from 400 ml of food extract (100 g of food) after mixing for 2 h at 4 C. The Sepharose-antibody-toxin complex was washed with 0.02 M phosphate-buffered saline at pH 7.2, and the toxin was dissociated by 2 to 4 ml of 0.2 M HCl-glycine plus 0.5 M NaCl buffer at pH 2.8. The recovered enterotoxin was free of interfering food components and could be detected serologically. Work to couple antibodies A, B, C, D, and E to Sepharose to recover all five toxins in one step is under study.
从食品中提取、浓缩和血清学检测葡萄球菌肠毒素既费力又耗时。通过将食品提取物与标记有特异性抗肠毒素B的不溶性基质接触,我们能够以灵敏且快速的方式从食品中回收毒素。在室温下将试剂混合2小时后,在pH 8.5(含0.5 M NaCl的0.1 M碳酸盐缓冲液)条件下,将免疫球蛋白G抗体连接到溴化氰活化的琼脂糖4B上。琼脂糖-抗体复合物(1 ml)在4℃混合2小时后,可从400 ml食品提取物(100 g食品)中特异性回收0.1至30 μg的肠毒素B。用pH 7.2的0.02 M磷酸盐缓冲盐水洗涤琼脂糖-抗体-毒素复合物,并用2至4 ml pH 2.8的0.2 M HCl-甘氨酸加0.5 M NaCl缓冲液解离毒素。回收的肠毒素不含干扰性食品成分,可进行血清学检测。将抗体A、B、C、D和E与琼脂糖偶联以一步回收所有五种毒素的工作正在研究中。