TY - JOUR
T1 - Development of an in vitro bioassay for Clostridium botulinum type B neurotoxin in foods that is more sensitive than the mouse bioassay
AU - Wictome, Matthew
AU - Newton, Kirsti
AU - Jameson, Karen
AU - Hallis, Bassam
AU - Dunnigan, Paul
AU - Mackay, Eric
AU - Clarke, Sally
AU - Taylor, Richard
AU - Gaze, Joy
AU - Foster, Keith
AU - Shone, Clifford
PY - 1999/9
Y1 - 1999/9
N2 - A novel, in vitro bioassay for detection of the botulinum type B neurotoxin in a range of media was developed. The assay is amplified by the enzymic activity of the neurotoxin's light chain and includes the following three stages: first, a small, monoclonal antibody-based immunoaffinity column captures the toxin; second, a peptide substrate is cleaved by using the endopeptidase activity of the type B neurotoxin; and finally, a modified enzyme-linked immunoassay system detects the peptide cleavage products. The assay is highly specific for type B neurotoxin and is capable of detecting type B toxin at a concentration of 5 pg ml-1 (0.5 mouse 50% lethal dose ml-1) in approximately 5 h. The format of the test was found to be suitable for detecting botulinum type B toxin in a range of foodstuffs with a sensitivity that exceeds the sensitivity of the mouse assay. Using highly specific monoclonal antibodies as the capture phase, we found that the endopeptidase assay was capable of differentiating between the type B neurotoxins produced by proteolytic and nonproteolytic strains of Clostridium botulinum type B.
AB - A novel, in vitro bioassay for detection of the botulinum type B neurotoxin in a range of media was developed. The assay is amplified by the enzymic activity of the neurotoxin's light chain and includes the following three stages: first, a small, monoclonal antibody-based immunoaffinity column captures the toxin; second, a peptide substrate is cleaved by using the endopeptidase activity of the type B neurotoxin; and finally, a modified enzyme-linked immunoassay system detects the peptide cleavage products. The assay is highly specific for type B neurotoxin and is capable of detecting type B toxin at a concentration of 5 pg ml-1 (0.5 mouse 50% lethal dose ml-1) in approximately 5 h. The format of the test was found to be suitable for detecting botulinum type B toxin in a range of foodstuffs with a sensitivity that exceeds the sensitivity of the mouse assay. Using highly specific monoclonal antibodies as the capture phase, we found that the endopeptidase assay was capable of differentiating between the type B neurotoxins produced by proteolytic and nonproteolytic strains of Clostridium botulinum type B.
UR - http://www.scopus.com/inward/record.url?scp=0032881904&partnerID=8YFLogxK
U2 - 10.1128/aem.65.9.3787-3792.1999
DO - 10.1128/aem.65.9.3787-3792.1999
M3 - Article
C2 - 10473376
AN - SCOPUS:0032881904
VL - 65
SP - 3787
EP - 3792
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
SN - 0099-2240
IS - 9
ER -