Sub-femtomole detection of 16s rRNA from Legionella pneumophila using surface plasmon resonance imaging

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DOIResolve DOI: http://doi.org/10.1016/j.bios.2013.08.032
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TypeArticle
Journal titleBiosensors and Bioelectronics
ISSN0956-5663
Volume52
Pages129135; # of pages: 7
Subject16S rRNA; Hybridization; Legionella pneumophila; Pathogen detection; Quantum dot; Surface plasmon resonance imaging; Bioassay; Pathogens; Plasmons; Signal processing; Surface plasmon resonance; RNA; RNA 16S; article; electron capture detection; fragmentation reaction; Legionella pneumophila; legionnaire disease; limit of detection; nonhuman; RNA sequence; sensitivity and specificity; surface plasmon resonance
AbstractLegionellosis has been and continues to be a life-threatening disease worldwide, even in developed countries. Given the severity and unpredictability of Legionellosis outbreaks, developing a rapid, highly specific, and sensitive detection method is thus of great pertinence. In this paper, we demonstrate that sub-femtomole levels of 16s rRNA from pathogenic Legionella pneumophila can be timely and effectively detected using an appropriate designed capture, detector probes, and a QD SPRi signal amplification strategy. To achieve specific and sensitive detection, optimal hybridization conditions and parameters were implemented. Among these parameters, fragmentation of the 16s rRNA and further signal amplification by QDs were found to be the main parameters contributing to signal enhancement. The appropriate design of the detector probes also increased the sensitivity of the detection system, mainly due to secondary structure of 16s rRNA. The use of 16s rRNA from L. pneumophila allowed for the detection of metabolically active pathogens with high sensitivity. Detection of 16s rRNA in solutions as diluted as 1. pM at 450. μL (0.45. femtomole) was achieved in less than 3. h, making our approach suitable for the direct, timely, and effective detection of L. pneumophila within man-made water systems. © 2013 Elsevier B.V.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC)
Peer reviewedYes
NPARC number21270852
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Record identifierc82122b1-a6c6-4939-881c-a41cb2ccc21d
Record created2014-02-17
Record modified2016-05-09
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