Gibson A M, Modi N K, Roberts T A, Shone C C, Hambleton P, Melling J
Agricultural and Food Research Council, Bristol Laboratory, Langford, UK.
J Appl Bacteriol. 1987 Sep;63(3):217-26. doi: 10.1111/j.1365-2672.1987.tb04939.x.
A monoclonal antibody-based amplified enzyme-linked immunosorbent assay (ELISA) method for detecting Clostridium botulinum type A toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5-4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated, 80 degrees C/5 min + 70 degrees C/2 h, before storage at 15 degrees, 20 degrees or 27 degrees C. The presence of specific toxin was confirmed by mouse bioassay and results compared with those of the amplified ELISA method. A total of 49 strains, 39 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 95 slurry samples were tested. Fourteen of 15 strains of type A Cl. botulinum and 34 of 36 slurry samples containing type A toxin were positive by ELISA. No false positive reactions occurred with Cl. botulinum types B, C, D, E and F, or with the 10 strains of Cl. sporogenes. However, toxin produced by one strain of Cl. botulinum type A (NCTC 2012) was not detected by the amplified ELISA.
评估了一种基于单克隆抗体的扩增酶联免疫吸附测定（ELISA）方法检测A型肉毒梭菌毒素的能力，该方法用于检测纯培养物上清液中的毒素以及接种到猪肉匀浆中的肉毒梭菌孢子生长后的毒素。含有NaCl（1.5 - 4.5% w/v）和多聚磷酸盐（0.3% w/v）的匀浆在15℃、20℃或27℃储存前，要么不加热，要么加热（80℃/5分钟 + 70℃/2小时）。通过小鼠生物测定法确认特定毒素的存在，并将结果与扩增ELISA方法的结果进行比较。总共测试了49株菌株，其中39株肉毒梭菌和10株生孢梭菌（腐败厌氧菌），以及95个匀浆样品。15株A型肉毒梭菌中的14株以及36个含有A型毒素的匀浆样品中的34个通过ELISA检测呈阳性。B、C、D、E和F型肉毒梭菌或10株生孢梭菌均未出现假阳性反应。然而，扩增ELISA未检测到一株A型肉毒梭菌（NCTC 2012）产生的毒素。
