High Sensitivity Detection of Active Botulinum Neurotoxin (BoNT) by Glyco-qPCR
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Authors
Kwon, S.J.
Jeong, E.J.
Yoo, Y.C.
Yang, G.-H.
Lee, J.C.
Dordick, J.S.
Linhardt, Robert J.
Lee, K.B.
Issue Date
2014
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
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.
Description
Analytical Chemistry, 86, 2279−2284
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
High Sensitivity Detection of Active Botulinum Neurotoxin (BoNT) by Glyco-qPCR," S. J. Kwon, E.J. Jeong, Y.C. Yoo, G.-H. Yang, J.C. Lee, J.S. Dordick, R.J. Linhardt, K.B. Lee, Analytical Chemistry, 86, 2279−2284, 2014.
Publisher
American Chemical Society (ACS)