Abstract

In this work, the use of laser-induced breakdown spectroscopy (LIBS) to differentiate live pathogens and killed viruses on substrates is investigated. Live pathogens B. anthracis Sterne strain and F. tularensis live vaccine strain were interrogated as lawn and colonies on agar; dilutions on agar; and dilutions on glass slides, and it was found possible to differentiate among all samples. UV killed hantavirus strains were studied as dilutions on slides and it was also found possible to differentiate among strains. To the best of our knowledge, this is the first study in which LIBS has been used to differentiate virus samples.

© 2012 Optical Society of America

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References

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  1. K. H. Esbensen, Multivariate Data Analysis—In Practice, 5th ed. (Camo, 1994).
  2. J. L. Gottfried, F. C. De Lucia, C. A. Munson, and A. W. Miziolek, “Standoff detection of chemical and biological threats using LIBS,” Appl. Spectrosc. 62, 353–363 (2008).
    [CrossRef]
  3. J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy,” J. Appl. Phys. 102, 014702 (2007).
    [CrossRef]
  4. J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Escherichia coli identification and strain discrimination using nanosecond laser-induced breakdown spectroscopy,” Appl. Phys. Lett. 90, 163901 (2007).
    [CrossRef]
  5. R. A. Multari, D. A. Cremers, J. M. Dupre, and J. E. Gustafson, “The use of laser-induced breakdown spectroscopy for distinguishing between bacterial pathogen species and strains,” Appl. Spectrosc. 64, 750–759 (2010).
    [CrossRef]
  6. Q. Mohaidat, S. Palchaudhuri, and S. J. Rehse, “The effect of bacterial environmental and metabolic stresses on a LIBS-based identification of Escherichia coli and Streptococcus viridans,” Appl. Spectrosc. 65, 386–392 (2011).
    [CrossRef]
  7. R. A. Multari and D. A. Cremers, “Methods for forming recognition algorithms for laser-induced breakdown spectroscopy,” U.S. patent application 12/981,626 (30Dec.2010).
  8. E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
    [CrossRef]
  9. J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).
  10. C. A. Knight, “The nature of some of the chemical differences among strains of tobacco mosaic virus,” presented in part before the Division of Biological Chemistry at the American Chemical Society at Atlantic City, 14–18April1947, http://www.jbc.org/content/171/1/297.full.pdf ; last accessed 11Nov.2011.

2011 (1)

2010 (1)

2008 (2)

J. L. Gottfried, F. C. De Lucia, C. A. Munson, and A. W. Miziolek, “Standoff detection of chemical and biological threats using LIBS,” Appl. Spectrosc. 62, 353–363 (2008).
[CrossRef]

E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
[CrossRef]

2007 (3)

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy,” J. Appl. Phys. 102, 014702 (2007).
[CrossRef]

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Escherichia coli identification and strain discrimination using nanosecond laser-induced breakdown spectroscopy,” Appl. Phys. Lett. 90, 163901 (2007).
[CrossRef]

Bondu-Hawkins, V. S.

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

Cremers, D. A.

De Lucia, F. C.

Diedrich, J.

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Escherichia coli identification and strain discrimination using nanosecond laser-induced breakdown spectroscopy,” Appl. Phys. Lett. 90, 163901 (2007).
[CrossRef]

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy,” J. Appl. Phys. 102, 014702 (2007).
[CrossRef]

Dupre, J. M.

Esbensen, K. H.

K. H. Esbensen, Multivariate Data Analysis—In Practice, 5th ed. (Camo, 1994).

Gias, E.

E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
[CrossRef]

Gottfried, J. L.

Gustafson, J. E.

Hall, P. R.

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

Hjelle, B.

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

Miziolek, A. W.

Mohaidat, Q.

Morgan, L. A. F.

E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
[CrossRef]

Multari, R. A.

Munson, C. A.

Nielsen, S. U.

E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
[CrossRef]

Palchaudhuri, S.

Q. Mohaidat, S. Palchaudhuri, and S. J. Rehse, “The effect of bacterial environmental and metabolic stresses on a LIBS-based identification of Escherichia coli and Streptococcus viridans,” Appl. Spectrosc. 65, 386–392 (2011).
[CrossRef]

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Escherichia coli identification and strain discrimination using nanosecond laser-induced breakdown spectroscopy,” Appl. Phys. Lett. 90, 163901 (2007).
[CrossRef]

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy,” J. Appl. Phys. 102, 014702 (2007).
[CrossRef]

Prescott, J. B.

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

Rehse, S. J.

Q. Mohaidat, S. Palchaudhuri, and S. J. Rehse, “The effect of bacterial environmental and metabolic stresses on a LIBS-based identification of Escherichia coli and Streptococcus viridans,” Appl. Spectrosc. 65, 386–392 (2011).
[CrossRef]

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Escherichia coli identification and strain discrimination using nanosecond laser-induced breakdown spectroscopy,” Appl. Phys. Lett. 90, 163901 (2007).
[CrossRef]

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy,” J. Appl. Phys. 102, 014702 (2007).
[CrossRef]

Toms, G. L.

E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
[CrossRef]

Ye, C.

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

Appl. Phys. Lett. (1)

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Escherichia coli identification and strain discrimination using nanosecond laser-induced breakdown spectroscopy,” Appl. Phys. Lett. 90, 163901 (2007).
[CrossRef]

Appl. Spectrosc. (3)

J. Appl. Phys. (1)

J. Diedrich, S. J. Rehse, and S. Palchaudhuri, “Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy,” J. Appl. Phys. 102, 014702 (2007).
[CrossRef]

J. Immunol. (1)

J. B. Prescott, P. R. Hall, V. S. Bondu-Hawkins, C. Ye, and B. Hjelle, “Early innate immune responses to Sin Nombre Hantavirus occur independently of IFN regulatory factor 3, characterized pattern recognition receptors and viral entry,” J. Immunol. 179, 1796–1802 (2007).

J. Virol. Methods (1)

E. Gias, S. U. Nielsen, L. A. F. Morgan, and G. L. Toms, “Purification of human respiratory syncytial virus by ultracentrifugation in iodixanol density gradient,” J. Virol. Methods 147, 328–332 (2008).
[CrossRef]

Other (3)

R. A. Multari and D. A. Cremers, “Methods for forming recognition algorithms for laser-induced breakdown spectroscopy,” U.S. patent application 12/981,626 (30Dec.2010).

C. A. Knight, “The nature of some of the chemical differences among strains of tobacco mosaic virus,” presented in part before the Division of Biological Chemistry at the American Chemical Society at Atlantic City, 14–18April1947, http://www.jbc.org/content/171/1/297.full.pdf ; last accessed 11Nov.2011.

K. H. Esbensen, Multivariate Data Analysis—In Practice, 5th ed. (Camo, 1994).

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Figures (6)

Fig. 1.
Fig. 1.

Experimental setup used to collect LIBS spectra of pathogens and viruses. The samples were located inside a BSL-2 hood. LIBS emission was collected along the path of the laser light to remove parallax.

Fig. 2.
Fig. 2.

Illustration of the process used to create the differential models needed for sample screening. The best models are those in which there is a wide separation in the prediction values obtained for the two sample groups being differentiated as is shown in Section 3 for the differentiation of B. anthracis from LVS in dilution on glass slides. Example spectra used to create a differentiation model are shown on the left. On the top right, the two-dimensional score space plot for the resulting model is shown. In the lower right, the prediction value results (average 20 spectra) obtained for spectra reserved for testing the model are shown.

Fig. 3.
Fig. 3.

Prediction results for differentiation models built for the lawn samples when test spectra are input (left) and when unknown or “blind” spectra are input (right). The differential models in the first two rows were designed to differentiate uncontaminated agar samples from all other samples (including dilutions on agar), whereas the models in the rows below were designed to differentiate the B. anthracis sample and the LVS sample from all other samples.

Fig. 4.
Fig. 4.

Prediction results for differentiation models built for the dilution samples when test spectra are input (left) and when unknown or “blind” spectra are input (right). Note the predictive distribution was shifted for the unknown samples. This indicates sample variability was not successfully captured in the modeling. See text for a detailed discussion of the shifted distribution.

Fig. 5.
Fig. 5.

Prediction results for differentiation models built for the dilution on slide samples when test spectra are input (left) and when unknown or “blind” spectra are input (right). The differential model in the top row was designed to differentiate uncontaminated slide samples from all other samples, whereas the models in the rows below were designed to differentiate the B. anthracis dilution on slide sample from all other slide samples and the LVS dilution on slide sample from all other slide samples.

Fig. 6.
Fig. 6.

Prediction results for differentiation models built for the hantavirus dilution on slide samples when test spectra are input. In the top row, prediction results on test spectra for models designed to differentiate the blank slide with water and the slide with Iodixanol are presented. The remaining rows show test spectra results for the hantavirus dilution models. Once the blank slide, the Iodixanol, and the PUU samples were removed from the analysis, it was possible to build single models to differentiate the remaining samples.

Tables (1)

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Table 1. Summary of Samples Prepared by the UNMHSC for This Study

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