Detection and Differentiation of Avian Mycoplasmas by Surface-Enhanced Raman Spectroscopy Based on a Silver Nanorod Array

Hennigan, Suzanne L.; Driskell, Jeremy D.; Ferguson‐Noel, Naola; Dluhy, Richard A.; Zhao, Yiping; Tripp, Ralph A.; Krause, Duncan C.

doi:10.1128/aem.07419-11

Cited by 38 publications

(40 citation statements)

References 34 publications

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“…49 Other studies showed that our SERS-based methods had equivalent-or-better detection limits than qPCR for analysis of pathogens in complex clinical samples. 50 Therefore, based on our previous experience, we feel confident that the methods described here can be extended to analyze biologically complex mixtures.…”

Section: Discussionmentioning

confidence: 80%

Identification of Virulence Determinants in Influenza Viruses

et al. 2014

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To date there is no rapid method to screen for highly pathogenic avian influenza strains that may be indicators of future pandemics. We report here the first development of an oligonucleotide-based spectroscopic assay to rapidly and sensitively detect a N66S mutation in the gene coding for the PB1-F2 protein associated with increased virulence in highly pathogenic pandemic influenza viruses. 5’-thiolated ssDNA oligonucleotides were employed as probes to capture RNA isolated from six influenza viruses, three having N66S mutations, two without the N66S mutation, and one deletion mutant not encoding the PB1-F2 protein. Hybridization was detected without amplification or labeling using the intrinsic surfaced-enhanced Raman spectrum of the DNA-RNA complex. Multivariate analysis identified target RNA binding from non-complementary sequences with 100% sensitivity, 100% selectivity, and 100% correct classification in the test data set. These results establish that optical-based diagnostic methods are able to directly identify diagnostic indicators of virulence linked to highly pathogenic pandemic influenza viruses without amplification or labeling.

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Section: Discussionmentioning

confidence: 80%

Identification of Virulence Determinants in Influenza Viruses

et al. 2014

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“…9, 53 The current work is the first to use the LBL method to ensure preparation of whole, intact encapsulated mycoplasma cells for use in SERS classification studies. The high quality and reproducibility of the LBL SERS spectra, as seen in Figs.…”

Section: Resultsmentioning

confidence: 99%

“…9, 30 The high discriminatory ability of LBL SERS seen in this study is similar to that seen when using Ag nanorod-based SERS methods to analyze the spectra of human and avian mycoplasmas, albeit with lower root mean square errors. 9, 53 …”

Section: Resultsmentioning

confidence: 99%

Layer-by-layer polyelectrolyte encapsulation of Mycoplasma pneumoniae for enhanced Raman detection

Rivera-Betancourt

Sheppard

Krause

et al. 2014

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Mycoplasma pneumoniae is a major cause of respiratory disease in humans and accounts for as much as 20% of all community-acquired pneumonia. Existing mycoplasma diagnosis is primarily limited by the poor success rate at culturing the bacteria from clinical samples. There is a critical need to develop a new platform for mycoplasma detection that has high sensitivity, specificity, and expediency. Here we report the layer-by-layer (LBL) encapsulation of M. pneumoniae cells with Ag nanoparticles in a matrix of the polyelectrolytes poly(allylamine hydrochloride) (PAH) and poly(styrene sulfonate) (PSS). We evaluated nanoparticle encapsulated mycoplasma cells as a platform for the differentiation of M. pneumoniae strains using surface enhanced Raman scattering (SERS) combined with multivariate statistical analysis. Three separate M. pneumoniae strains (M129, FH and II-3) were studied. Scanning electron microscopy and fluorescence imaging showed that the Ag nanoparticles were incorporated between the oppositely charged polyelectrolyte layers. SERS spectra showed that LBL encapsulation provides excellent spectral reproducibility. Multivariate statistical analysis of the Raman spectra differentiated the three M. pneumoniae strains with 97 – 100% specificity and sensitivity, and low (0.1 – 0.4) root mean square error. These results indicated that nanoparticle and polyelectrolyte encapsulation of M. pneumoniae is a potentially powerful platform for rapid and sensitive SERS-based bacterial identification.

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“…In a more recent study, Raman spectral signatures of multiple strains of avian mycoplasma species including Acholeplasma laidlawii , Mycoplasma gallinarum , Mycoplasma gallinaceum , Mycoplasma synoviae , and M. gallisepticum , and vaccine strains 6/85, F, and ts-11 were collected and processed by multivariate analysis to differentiate laboratory cultures of mycoplasma species [79]. A PLS-DA model classified the spectra for all species with 93 to 100% sensitivity and with similar specificity for all species except Mycoplasma synoviae , which was distinguishable with only 80% specificity.…”

Section: Detection Of Bacteriamentioning

confidence: 99%

Ag nanorod based surface‐enhanced Raman spectroscopy applied to bioanalytical sensing

Negri

Dluhy

2012

Journal of Biophotonics

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Recent progress in substrate nanofabrication has led to the development of Ag nanorod arrays as uniform, reproducible, large area SERS-active substrates with high signal enhancement. These novel nanostructures fabricated by oblique angle vapor deposition (OAD) offer a robust platform for the rapid detection of biological agents and open new perspectives for the development and integration of biomedical diagnostic for clinical and therapeutic applications. Ag nanorod arrays have been investigated as SERS-active substrates for the detection and identification of pathogens, including bacteria and viruses, as well as to evaluate the potential of this biosensing platform for bio-recognition of high affinity events using oligonucleotide-modified substrates. This review summarizes the various nanostructured substrates designed for SERS-based applications, highlights the nanofabrication methodology used to produce Ag nanorod arrays, outlines their morphological and physical properties, and provides a summary of the most recent uses of these substrates for clinical diagnostic and biomedical applications.

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Detection and Differentiation of Avian Mycoplasmas by Surface-Enhanced Raman Spectroscopy Based on a Silver Nanorod Array

Cited by 38 publications

References 34 publications

Identification of Virulence Determinants in Influenza Viruses

Identification of Virulence Determinants in Influenza Viruses

Layer-by-layer polyelectrolyte encapsulation of Mycoplasma pneumoniae for enhanced Raman detection

Ag nanorod based surface‐enhanced Raman spectroscopy applied to bioanalytical sensing

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