title: Bioluminescence assay for the highly sensitive detection of botulinum neurotoxin A activity.
abstract: This article describes a novel bioluminescence assay for detecting the proteolytic activity of Botulinum NeuroToxins (BoNT) in complex matrices. The assay is capable of detecting traces of BoNT in blood samples as well as in food drinks. The assay was responsive to BoNT/A subtypes 1 to 5, and serotype E3 in buffered solutions. It was responsive to filtered Clostridium botulinum supernatants and BoNT/A1 in complex with neurotoxin associated proteins in bouillon and milk (3.8% fat) down to 400 fM after 4 h RT incubation and in bouillon at concentrations down to 120 fM after 21 h RT incubation. In combination with an immunocapture/enrichment step it could detect BoNT/A1 in citrated plasma at concentrations down to 30 fM (1.2 mouse LD50 per mL). The simplicity of the assay, combined with a demonstrated ability to lyophilize the reagents, demonstrates its usefulness for detection of BoNT in non-specialised analytical laboratories.
authors: Freiburg Materials Research Centre (FMF), Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Str. 21, 79104 Freiburg, Germany. Gregory.Stevens@fmf.uni-freiburg.de.
authors: G B
authors: GB
authors: Stevens
authors: D A
authors: DA
authors: Silver
authors: A
authors: A
authors: Zgaga-Griesz
authors: W G
authors: WG
authors: Bessler
authors: S K
authors: SK
authors: Vashist
authors: P
authors: P
authors: Patel
authors: K
authors: K
authors: Achazi
authors: J
authors: J
authors: Strotmeier
authors: S
authors: S
authors: Worbs
authors: M B
authors: MB
authors: Dorner
authors: B G
authors: BG
authors: Dorner
authors: D
authors: D
authors: Pauly
authors: A
authors: A
authors: Rummel
authors: G A
authors: GA
authors: Urban
authors: M
authors: M
authors: Krueger
mesh_terms: D000818:Animals; D019274:Botulinum Toxins, Type A / Q000032:analysis* / Q000737:chemistry*; D003014:Clostridium botulinum / Q000737:chemistry; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D017433:Protein Structure, Secondary
chemical_list: D019274:Botulinum Toxins, Type A