Cloth-based hybridization array system for the detection of Clostridium botulinum type A, B, E, and F neurotoxin genes.
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.
Gauthier
M
M
Ottawa Laboratory (Carling), Canadian Food Inspection Agency, Building 22, Central Experimental Farm, 960 Carling Avenue, Ottawa, Ontario, Canada K1A 0C6.
Cadieux
B
B
Austin
J W
JW
Blais
B W
BW
D001905:Botulinum Toxins / Q000235:genetics*; D019274:Botulinum Toxins, Type A / Q000235:genetics; D003014:Clostridium botulinum / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D005798:Genes, Bacterial; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D012703:Serotyping
D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A