Craven Kathy E, Ferreira Joseph L, Harrison Mark A, Edmonds Paul
U.S. Food and Drug Administration, Southeast Regional Laboratory, Atlanta, GA 30309, USA.
J AOAC Int. 2002 Sep-Oct;85(5):1025-8.
Clostridium botulinum organisms generally produce 1 of 4 neurotoxin types (A, B, E, and F) associated with human illness. Neurotoxin type determination is important in identification of the bacterium. A polymerase chain reaction (PCR) method was developed to identify 24 h botulinal cultures as potential types A, B, E, and F neurotoxin producers as well as other clostridial species which also produce neurotoxins. Components of the PCR and amplification conditions were adjusted for optimal amplification of toxin gene target regions to enable simultaneous testing for types A, B, E, and F in separate tubes using a single thermal cycler. Each primer set was specific for its corresponding toxin type. A DNA extraction procedure was also included to remove inhibitory substances that may affect amplification. This procedure is rapid, sensitive, and specific for identification of toxigenic C. botulinum.
肉毒梭菌通常产生与人类疾病相关的4种神经毒素类型（A、B、E和F）中的一种。神经毒素类型的确定对于该细菌的鉴定很重要。开发了一种聚合酶链反应（PCR）方法，以鉴定培养24小时的肉毒杆菌培养物是否为潜在的A、B、E和F型神经毒素产生者，以及其他也产生神经毒素的梭菌属物种。调整了PCR的成分和扩增条件，以实现毒素基因靶区域的最佳扩增，从而能够使用单个热循环仪在单独的试管中同时检测A、B、E和F型。每个引物组对其相应的毒素类型具有特异性。还包括一种DNA提取程序，以去除可能影响扩增的抑制性物质。该程序快速、灵敏且特异，可用于鉴定产毒肉毒梭菌。
