Cloth-Based Hybridization Array System for the Detection of Clostridium botulinum Type A, B, E, and F Neurotoxin Genes

Authors: Gauthier, M.1; Cadieux, B.2; Austin, J. W.2; Blais, B. W.1

Source: Journal of Food Protection®, Number 7, July 2005, pp. 1336-1534 , pp. 1477-1483(7)

Publisher: International Association for Food Protection

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Abstract:

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 digoxigenindUTP 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.

Document Type: Research Article

Affiliations: 1: Ottawa Laboratory (Carling), Canadian Food Inspection Agency, Building 22, Central Experimental Farm, 960 Carling Avenue, Ottawa, Ontario, Canada K1A 0C6 2: Microbiology Research Division, Bureau of Microbial Hazards, Food Directorate, Health Products and Food Branch, Health Canada, Ross Avenue, Postal Locator 2204A2, Ottawa, Ontario, Canada K1A 0L2

Publication date: July 1, 2005

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