GENE PROBE-BASED DETECTION OF TYPE-E BOTULINUM NEUROTOXIN BINDING-PROTEIN USING POLYMERASE CHAIN-REACTION

Using primers based on the nucleotide sequence of a neurotoxin binding protein from Type E Clostridium botulinum cultures, an amplified DNA product was obtained through polymerase chain reaction. The 400 base p air amplified DNA fragment was detectable with as low as 0.1 pg templa te DNA from Type E C. botulinum, and its fidelity was confirmed by Sou thern blotting using a DNA probe designed to detect the expected ampli fied DNA fragment. On the other hand, no DNA amplification was observe d with as high as 10 ng template DNA from related Types A and B C. bot ulinum or from C. tetani, indicating the specificity of the probe. Cop yright (C) 1996 Elsevier Science Ltd