Gauthier M, Cadieux B, Austin J W, Blais B W
Ottawa Laboratory (Carling), Canadian Food Inspection Agency, Building 22, Central Experimental Farm, 960 Carling Avenue, Ottawa, Ontario, Canada K1A 0C6.
J Food Prot. 2005 Jul;68(7):1477-83. doi: 10.4315/0362-028x-68.7.1477.
A simple cloth-based hybridization array system was developed for the characterization of Clostridium botulinum isolates based on the botulinum neurotoxin serotype. Bacterial isolates were subjected to a multiplex PCR incorporating digoxigenin-dUTP and primers targeting the four botulinum neurotoxin gene serotypes (A, B, E, and F) predominantly involved in human illness, followed by hybridization of the amplicons with an array of toxin gene-specific oligonucleotide probes immobilized on polyester cloth and subsequent immunoenzymatic assay of the bound digoxigenin label. This system provided sensitive and specific detection of the different botulinum neurotoxin gene markers in a variety of C. botulinum strains, exhibiting the expected patterns of reactivity with a panel of target and nontarget organisms.
开发了一种基于布的简单杂交阵列系统，用于根据肉毒杆菌神经毒素血清型对肉毒杆菌分离株进行表征。将细菌分离株进行多重PCR，该PCR掺入地高辛配基-dUTP和靶向主要涉及人类疾病的四种肉毒杆菌神经毒素基因血清型（A、B、E和F）的引物，然后将扩增产物与固定在聚酯布上的一系列毒素基因特异性寡核苷酸探针杂交，并随后对结合的地高辛配基标记进行免疫酶测定。该系统能够灵敏且特异地检测各种肉毒杆菌菌株中的不同肉毒杆菌神经毒素基因标记，与一组靶标和非靶标生物体呈现出预期的反应模式。
