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Clinical Utility of PNA-FISH for Burn Wound Diagnostics: A Noninvasive, Culture-Independent Technique for Rapid Identification of Pathogenic Organisms in Burn Wounds

Weaver, Alan J, Brandenburg, Kenneth S, Sanjar, Fatemeh, Wells, Adrienne R, Peacock, Trent J, Leung, Kai P·2019·Journal of Burn Care & Research
DOI 10.1093/jbcr/irz047PMC PMC6587406
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Figure 1. Validation of multiplexed PNA-FISH and CW on planktonic cell cultures. A. Tri-species planktonic cell cultures were stained to both validate the procedure and serve as controls for in vivo studies. The PNA-FISH probes showed specificity toward their respective bacteria. CW was specific to Candida albicans (blue); however, it also increased the fluorescence of the fungi in the green channel (arrowhead). B. Unstained monospecies cultures demonstrated faint autofluorescence of both Staphylococcus aureus and C. albicans in the green channel; however, no autofluorescence was seen from Pseudomonas aeruginosa in any channel. All images were taken with 63× objective.
Figure 1. Validation of multiplexed PNA-FISH and CW on planktonic cell cultures. A. Tri-species planktonic cell cultures were stained to both validate the procedure and serve as controls for in vivo studies. The PNA-FISH probes showed specificity toward their respective bacteria. CW was specific to Candida albicans (blue); however, it also increased the fluorescence of the fungi in the green channel (arrowhead). B. Unstained monospecies cultures demonstrated faint autofluorescence of both Staphylococcus aureus and C. albicans in the green channel; however, no autofluorescence was seen from Pseudomonas aeruginosa in any channel. All images were taken with 63× objective.CC BY-NC · PubMed Central
Figure 2. Pathogenic organisms identified from swabs of partial-thickness burn wounds. Each panel represents an individual channel and location for visualization of the microorganisms. Pseudomonas aeruginosa was clearly seen in the red channel as bright clusters of rods with minimal staining of the tissue debris. In the green channel, there were several clusters of Staphylococcus aureus that could be distinguished from the autofluorescent nature of the tissue debris. Tissue debris autofluorescence was also seen in the blue channel (not shown in panel), but Candida albicans was still clearly identifiable in both hyphae and bud form (inset, same scale). All images were taken with 63× objective. Single scale bar applies to all panels.
Figure 2. Pathogenic organisms identified from swabs of partial-thickness burn wounds. Each panel represents an individual channel and location for visualization of the microorganisms. Pseudomonas aeruginosa was clearly seen in the red channel as bright clusters of rods with minimal staining of the tissue debris. In the green channel, there were several clusters of Staphylococcus aureus that could be distinguished from the autofluorescent nature of the tissue debris. Tissue debris autofluorescence was also seen in the blue channel (not shown in panel), but Candida albicans was still clearly identifiable in both hyphae and bud form (inset, same scale). All images were taken with 63× objective. Single scale bar applies to all panels.CC BY-NC · PubMed Central
Figure 3. PNA-FISH and CW treatment of histological sections reveals pathogenic niches throughout the burn wound. Overlays of the blue channel with the red (A, B) and green (C, D) channels for unstained (A, C) and stained (B, D) tissue. Unstained sections showed autofluorescence in the red and green channels, as wells as voids that resembled Candida albicans (white arrowheads), which could be seen throughout the stained sections (yellow arrowheads). Red Pseudomonas aeruginosa rods (B, dagger) and green Staphylococcus aureus cocci (D, asterisk) were also seen intermingled with C. albicans. All images were taken with 63× objective.
Figure 3. PNA-FISH and CW treatment of histological sections reveals pathogenic niches throughout the burn wound. Overlays of the blue channel with the red (A, B) and green (C, D) channels for unstained (A, C) and stained (B, D) tissue. Unstained sections showed autofluorescence in the red and green channels, as wells as voids that resembled Candida albicans (white arrowheads), which could be seen throughout the stained sections (yellow arrowheads). Red Pseudomonas aeruginosa rods (B, dagger) and green Staphylococcus aureus cocci (D, asterisk) were also seen intermingled with C. albicans. All images were taken with 63× objective.CC BY-NC · PubMed Central

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Metadata

Authors
Weaver, A. J., Brandenburg, K. S., Sanjar, F., Wells, A. R., Peacock, T. J., & Leung, K. P.
Venue
Journal of Burn Care & Research
Publisher
Oxford University Press (OUP)
Published
2019-03-20
Volume
40
Issue
4
Pages
464-470
ISSN
  • 1559-047X
  • 1559-0488
DOI
10.1093/jbcr/irz047
PMC
PMC6587406
Cited by
12
References
0
Language
en
Licence
creativecommons.org/licenses/by-nc/4.0/ (vor)
Funders
  • Intramural Program (G_001-2016-USAISR)
  • Combat Casualty Care Research Directorate
  • U.S. Army Medical Research and Materiel Command (MRMC), Naval Medical Research Center’s Advanced Medical Development Program (N3239815MHX040)
  • Oak Ridge Institute for Science and Education

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CoChat. (2026). Citation check: Clinical Utility of PNA-FISH for Burn Wound Diagnostics: A Noninvasive, Culture-Independent Technique for Rapid Identification of Pathogenic Organisms in Burn Wounds. https://app.cochat.ai/papers/10.1093/jbcr/irz047
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Weaver, A. J., Brandenburg, K. S., Sanjar, F., Wells, A. R., Peacock, T. J., & Leung, K. P. (2019). Clinical Utility of PNA-FISH for Burn Wound Diagnostics: A Noninvasive, Culture-Independent Technique for Rapid Identification of Pathogenic Organisms in Burn Wounds. Journal of Burn Care & Research. https://doi.org/10.1093/jbcr/irz047