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Real-time electrochemical detection of pathogen DNA using electrostatic interaction of a redox probe.
MedLine Citation:
PMID:  23230566     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Electrostatic redox probes interaction has been widely rendered for DNA quantification. We have established a proof-of-principle by using the ruthenium hexaamine molecule [Ru(NH(3))(6)](3+). We have applied this method for real-time electrochemical monitoring of a loop mediated isothermal amplification (LAMP) amplicon of target genes of Escherichia coli and Staphylococcus aureus by square wave voltammetry (SWV). Ruthenium hexaamine interaction with free DNAs in solution without being immobilized onto the biochip surface enabled us to discard the time-consuming overnight probe immobilization step in DNA quantification. We have measured the changes in the cathodic current signals using screen printed low-cost biochips both in the presence and the absence of LAMP amplicons of target DNAs in the solution-phase. By using this novel probe, we successfully carried out the real-time isothermal amplification and detection in less than 30 min for S. aureus and E. coli with a sensitivity up to 30 copies μL(-1) and 20 copies μL(-1), respectively. The cathode peak height of the current was related to the extent of amplicon formation and the amount of introduced template genomic DNA. Importantly, since laborious probe immobilization is not necessary at all, and both the in vitro amplification and real-time monitoring are performed in a single polypropylene tube using a single biochip, this novel approach could avoid all potential cross-contamination in the whole procedure.
Authors:
Minhaz Uddin Ahmed; Sharifun Nahar; Mohammadali Safavieh; Mohammed Zourob
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-10
Journal Detail:
Title:  The Analyst     Volume:  -     ISSN:  1364-5528     ISO Abbreviation:  Analyst     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0372652     Medline TA:  Analyst     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Institut National de la Recherche Scientifique (INRS) - Énergie Matériaux Télécommunications (EMT), 1650 Boulevard Lionel-Boulet, Varennes, J3X 1S2, Quebec, Canada.
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