Issue 3, 2014

Simultaneous detection and quantification of three bacterial meningitis pathogens by SERS

Abstract

Bacterial meningitis is well known for its rapid onset and high mortality rates, therefore rapid detection of bacteria found in cerebral spinal fluid (CSF) and subsequent effective treatment is crucial. A new quantitative assay for detection of three pathogens that result in bacterial meningitis using a combination of lambda exonuclease (λ-exonuclease) and surface enhanced Raman scattering (SERS) is reported. SERS challenges current fluorescent-based detection methods in terms of both sensitivity and more importantly the detection of multiple components in a mixture, which is becoming increasingly more desirable for clinical diagnostics. λ-Exonuclease is a processive enzyme that digests one strand of double stranded DNA bearing a terminal 5′-phosphate group. The new assay format involves the simultaneous hybridisation of two complementary DNA probes (one containing a SERS active dye) to a target sequence followed by λ-exonuclease digestion of double stranded DNA and SERS detection of the digestion product. Three meningitis pathogens were successfully quantified in a multiplexed test with calculated limits of detection in the pico-molar range, eliminating the need for time consuming culture based methods that are currently used for analysis. Quantification of each individual pathogen in a mixture using SERS is complex, however, this is the first report that this is possible using the unique spectral features of the SERS signals combined with partial least squares (PLS) regression. This is a powerful demonstration of the ability of this SERS assay to be used for analysis of clinically relevant targets with significant advantages over existing approaches and offers the opportunity for future deployment in healthcare applications.

Graphical abstract: Simultaneous detection and quantification of three bacterial meningitis pathogens by SERS

Article information

Article type
Edge Article
Submitted
16 Oct 2013
Accepted
02 Dec 2013
First published
20 Dec 2013
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2014,5, 1030-1040

Author version available

Simultaneous detection and quantification of three bacterial meningitis pathogens by SERS

K. Gracie, E. Correa, S. Mabbott, J. A. Dougan, D. Graham, R. Goodacre and K. Faulds, Chem. Sci., 2014, 5, 1030 DOI: 10.1039/C3SC52875H

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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