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.
| Language | English |
|---|---|
| Pages | 1030-1040 |
| Number of pages | 11 |
| Journal | Chemical Science |
| Volume | 5 |
| Issue number | 3 |
| Early online date | 20 Dec 2013 |
| DOIs | |
| Publication status | Published - 1 Mar 2014 |
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Keywords
- bacterial meningitis
- cerebral spinal fluid
- surface enhanced Raman scattering
- meningitis pathogens
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}
Simultaneous detection and quantification of three bacterial meningitis pathogens by SERS. / Gracie, Kirsten; Correa, Elon; Mabbott, Samuel; Dougan, Jennifer A.; Graham, Duncan; Goodacre, Royston; Faulds, Karen.
In: Chemical Science, Vol. 5, No. 3, 01.03.2014, p. 1030-1040.Research output: Contribution to journal › Article
TY - JOUR
T1 - Simultaneous detection and quantification of three bacterial meningitis pathogens by SERS
AU - Gracie, Kirsten
AU - Correa, Elon
AU - Mabbott, Samuel
AU - Dougan, Jennifer A.
AU - Graham, Duncan
AU - Goodacre, Royston
AU - Faulds, Karen
PY - 2014/3/1
Y1 - 2014/3/1
N2 - 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.
AB - 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.
KW - bacterial meningitis
KW - cerebral spinal fluid
KW - surface enhanced Raman scattering
KW - meningitis pathogens
UR - http://www.scopus.com/inward/record.url?scp=84893476306&partnerID=8YFLogxK
U2 - 10.1039/c3sc52875h
DO - 10.1039/c3sc52875h
M3 - Article
VL - 5
SP - 1030
EP - 1040
JO - Chemical Science
T2 - Chemical Science
JF - Chemical Science
SN - 2041-6520
IS - 3
ER -