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2016-06-29Zeitschriftenartikel DOI: 10.1039/C6AN00693K
Detection, differentiation, and identification of botulinum neurotoxin serotypes C, CD, D, and DC by highly specific immunoassays and mass spectrometry
dc.contributor.authorHansbauer, Eva-Maria
dc.contributor.authorSkiba, Martin
dc.contributor.authorEndermann, Tanja
dc.contributor.authorWeisemann, Jasmin
dc.contributor.authorStern, Daniel
dc.contributor.authorDorner, Martin
dc.contributor.authorFinkenwirth, Friedrich
dc.contributor.authorWolf, Jessica
dc.contributor.authorLuginbühl, Werner
dc.contributor.authorMesselhäußer, Ute
dc.contributor.authorBellanger, Laurent
dc.contributor.authorWoudstra, Cédric
dc.contributor.authorRummel, Andreas
dc.contributor.authorFach, Patrick
dc.contributor.authorDorner, Brigitte
dc.date.accessioned2018-05-07T20:19:53Z
dc.date.available2018-05-07T20:19:53Z
dc.date.created2017-08-02
dc.date.issued2016-06-29none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reCZZXtIyjt8U/PDF/27FLfbJObPTtg.pdf
dc.identifier.urihttp://edoc.rki.de/176904/2740
dc.description.abstractBotulinum neurotoxin (BoNT) serotypes C and D and their mosaic variants CD and DC cause severe cases of botulism in animal husbandry and wildlife. Epidemiological data on the exact serotype or toxin variant causing outbreaks are rarely available, mainly because of their high sequence identity and the lack of fast and specific screening tools to detect and differentiate the four similar toxins. To fill this gap, we developed four highly specific sandwich enzyme-linked immunosorbent assays (ELISAs) able to detect and differentiate botulinum neurotoxins type BoNT/C, D, CD, and DC based on four distinct combinations of specific monoclonal antibodies targeting both conserved and divergent subdomains of the four toxins. Here, highly sensitive detection with detection limits between 2 and 24 pg mL−1 was achieved. The ELISAs were extensively validated and results were compared with data obtained by quantitative real-time PCR using a panel of Clostridium botulinum strains, real sample materials from veterinary botulism outbreaks, and non-BoNT-producing Clostridia. Additionally, in order to verify the results obtained by ELISA screening, the new monoclonal antibodies were used for BoNT enrichment and subsequent detection (i) on a functional level by endopeptidase mass spectrometry (Endopep-MS) assays and (ii) on a protein sequence level by LC-MS/MS spectrometry. Based on all technical information gathered in the validation study, the four differentiating ELISAs turned out to be highly reliable screening tools for the rapid analysis of veterinary botulism cases and should aid future field investigations of botulism outbreaks and the acquisition of epidemiological data.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Biologische Sicherheit
dc.subject.ddc610 Medizin
dc.titleDetection, differentiation, and identification of botulinum neurotoxin serotypes C, CD, D, and DC by highly specific immunoassays and mass spectrometry
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10054150
dc.identifier.doi10.1039/C6AN00693K
dc.identifier.doihttp://dx.doi.org/10.25646/2665
local.edoc.container-titleThe Analyst
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.pubs.rsc.org/en/journals/journal/an
local.edoc.container-publisher-nameRoyal Society of Chemistry
local.edoc.container-volume141
local.edoc.container-issue18
local.edoc.container-year2016

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