title: High sensitivity detection of active botulinum neurotoxin by glyco-quantitative polymerase chain-reaction.
abstract: The sensitive detection of highly toxic botulinum neurotoxin (BoNT) from Clostridium botulinum is of critical importance because it causes human illnesses if foodborne or introduced in wounds and as an iatrogenic substance. Moreover, it has been recently considered a possible biological warfare agent. Over the past decade, significant progress has been made in BoNT detection technologies, including mouse lethality assays, enzyme-linked immunosorbent assays, and endopeptidase assays and by mass spectrometry. Critical assay requirements, including rapid assay, active toxin detection, sensitive and accurate detection, still remain challenging. Here, we present a novel method to detect active BoNTs using a Glyco-quantitative polymerase chain-reaction (qPCR) approach. Sialyllactose, which interacts with the binding-domain of BoNTs, is incorporated into a sialyllactose-DNA conjugate as a binding-probe for active BoNT and recovered through BoNT-immunoprecipitation. Glyco-qPCR analysis of the bound sialyllactose-DNA is then used to detect low attomolar concentrations of BoNT and attomolar to femtomolar concentrations of BoNT in honey, the most common foodborne source of infant botulism.
authors: Department of Chemical and Biological Engineering, Department of Chemistry and Chemical Biology, and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute , Troy, New York 12180, United States.
authors: Seok Joon
authors: SJ
authors: Kwon
authors: Eun Ji
authors: EJ
authors: Jeong
authors: Yung Choon
authors: YC
authors: Yoo
authors: Chao
authors: C
authors: Cai
authors: Gi-Hyeok
authors: GH
authors: Yang
authors: Jae Chul
authors: JC
authors: Lee
authors: Jonathan S
authors: JS
authors: Dordick
authors: Robert J
authors: RJ
authors: Linhardt
authors: Kyung Bok
authors: KB
authors: Lee
mesh_terms: D001905:Botulinum Toxins / Q000032:analysis*; D057230:Limit of Detection; D013058:Mass Spectrometry; D009498:Neurotoxins / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D009498:Neurotoxins; D001905:Botulinum Toxins