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Applied and Environmental Microbiology, December 2005, p. 7897-7903, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7897-7903.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Sensitive Detection of Botulinum Neurotoxin Types C and D with an Immunoaffinity Chromatographic Column Test

Frank Gessler,* Katrin Hampe, and Helge Böhnel

Institute for Applied Biotechnology in the Tropics, Kellnerweg 6, 37077 Göttingen, Germany

Received 29 December 2004/ Accepted 17 August 2005

A sensitive and specific immunoassay for the simultaneous detection of Clostridium botulinum type C (BoNT/C) and type D neurotoxin was developed. Goat anti-mouse immunoglobulin G was bound to polyethylene disks in a small disposable column used for this assay. The sample was preincubated together with monoclonal antibodies specific for the heavy chain of BoNT/C and D and affinity-purified, biotinylated polyclonal antibodies against these neurotoxins. This complex was captured on the assay disk. Streptavidin-poly-horseradish peroxidase was used as a conjugate, and a precipitating substrate allowed the direct semiquantitative readout of the assay, if necessary. For a more accurate quantitative detection, the substrate can be eluted and measured in a photometer. Depending on the preincubation time, a sensitivity of 1 mouse lethal dose ml–1 was achieved in culture supernatants.

* Corresponding author. Mailing address: Institute for Applied Biotechnology in the Tropics, Kellnerweg 6, 37077 Göttingen, Germany. Phone: 49 (0)551 393393. Fax: 49 (0)551 393408. E-mail: fgessle{at}gwdg.de.

Applied and Environmental Microbiology, December 2005, p. 7897-7903, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7897-7903.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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