Ferreira Joseph L, Maslanka Susan, Johnson Eric, Goodnough Mike
U.S. Food and Drug Administration, 60 8th St. NE, Atlanta, GA 30309, USA.
J AOAC Int. 2003 Mar-Apr;86(2):314-31.
An amplified enzyme-linked immunosorbent assay (amp-ELISA) was compared with the AOAC Official Method 977.26 for detection of Clostridium botulinum and its toxins in foods. Eleven laboratories participated and the results of 10 laboratories were used in the study. Two anaerobic culture media, tryptone peptone glucose yeast extract (TPGY) and cooked meat medium (CMM) were used to generate toxic samples with types A, B, E, and F botulinal strains. Nonbotulinal clostridia were also tested. The toxicity of each botulinal culture was determined by the AOAC method, and the cultures were then diluted, if necessary, to high (about 10,000 minimal lethal dose [MLD]/mL) and low (about 100 MLD/mL) test samples. The overall sensitivity of detection in TPGY and CMM cultures with the amp-ELISA was 94.7% at about 100 MLD/mL and 99.6% for samples with > or = 10,000 MLD/mL toxicity. The amp-ELISA detection sensitivity for low toxin samples was 92.3% in TPGY and 99.4% in CMM. The false-positive rate ranged from 1.5% for type A to 28.6% for type F in TPGY, and from 2.4% for type A to 11.4% for type F in CMM. Most of the cross-reactivity was due to detection of other botulinal types, especially in high toxin samples. The amp-ELISA could be used to screen suspect cultures for botulinal toxins. Positive amp-ELISA samples would be confirmed by the AOAC reference method.
将一种扩增酶联免疫吸附测定法（amp-ELISA）与美国官方分析化学师协会（AOAC）的官方方法977.26进行比较，以检测食品中的肉毒梭菌及其毒素。11个实验室参与了研究，本研究采用了其中10个实验室的结果。使用两种厌氧培养基，即胰蛋白胨葡萄糖酵母提取物（TPGY）培养基和庖肉培养基（CMM），用A、B、E和F型肉毒杆菌菌株制备有毒样品。还对非肉毒梭菌进行了检测。每种肉毒杆菌培养物的毒性通过AOAC方法测定，然后根据需要将培养物稀释为高浓度（约10000最小致死剂量[MLD]/mL）和低浓度（约100 MLD/mL）的测试样品。amp-ELISA检测TPGY和CMM培养物中约100 MLD/mL样品的总体灵敏度为94.7%，毒性≥10000 MLD/mL样品的灵敏度为99.6%。amp-ELISA对TPGY中低毒素样品的检测灵敏度为92.3%，对CMM中低毒素样品的检测灵敏度为99.4%。TPGY中假阳性率从A型的1.5%到F型的28.6%不等，CMM中假阳性率从A型的2.4%到F型的11.4%不等。大多数交叉反应是由于检测到其他肉毒杆菌类型，尤其是在高毒素样品中。amp-ELISA可用于筛查可疑培养物中的肉毒杆菌毒素。amp-ELISA检测呈阳性的样品将通过AOAC参考方法进行确认。
