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Home / Equine Research Bank / PLoS One / A real-time impedance based method to assess Rhodococcus equ...
PloS one2013; 8(3); e60612; doi: 10.1371/journal.pone.0060612

A real-time impedance based method to assess Rhodococcus equi virulence.

Abstract: Rhodococcus equi is a facultative intracellular pathogen of macrophages and the causative agent of foal pneumonia. R. equi virulence is usually assessed by analyzing intracellular growth in macrophages by enumeration of bacteria following cell lysis, which is time consuming and does not allow for a high throughput analysis. This paper describes the use of an impedance based real-time method to characterize proliferation of R. equi in macrophages, using virulent and attenuated strains lacking the vapA gene or virulence plasmid. Image analysis suggested that the time-dependent cell response profile (TCRP) is governed by cell size and roundness as well as cytoxicity of infecting R. equi strains. The amplitude and inflection point of the resulting TCRP were dependent on the multiplicity of infection as well as virulence of the infecting strain, thus distinguishing between virulent and attenuated strains.
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Publication Date: 2013-03-28 PubMed ID: 23555995PubMed Central: PMC3610927DOI: 10.1371/journal.pone.0060612Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study introduces a real-time technique for assessing the virulence of Rhodococcus equi, a microbe responsible for lung infection in foals, based on its multiplicative behavior within macrophages.

Objective of the Study

  • The study aimed to explore a real-time, efficient method for assessing the virulence of Rhodococcus equi, a bacteria that causes pneumonia in foals. Traditionally, this assessment is performed by evaluating its intracellular growth in macrophages – a process that is painstakingly long and doesn’t support high throughput analysis.

Methodology

  • Pathogenicity of R. equi inside macrophages was observed using an impedance-based approach in real-time.
  • The study compared the growth of both virulent and attenuated (weakened or less harmful) forms of these bacteria.
  • Attenuated strains lacked the vapA gene or virulence plasmid which are integral to their pathogenicity.
  • Image analysis was used to understand that the time-dependent cell response profile (TCRP) was regulated by the size and roundness of cells, and the cytotoxicity of the infecting strains.

Observed Findings

  • The amplitude and the inflection point of the resulting TCRP were directly influenced by the multiplicity of infection (the number of infecting bacteria entering a single cell) as well as the virulence of the infecting strain.
  • The study established that this can differentiate between the virulent and attenuated strains successfully.

Significance of the Study

  • The comprehensive analysis of the parasite within the host cell using this novel approach provides a streamlined method for virulence assessment.
  • Since this method is less time-consuming than traditional tests and supports high throughput analysis, it could be helpful in large-scale or frequent screenings.

Cite This Article

APA
Miranda-CasoLuengo AA, Miranda-CasoLuengo R, Lieggi NT, Luo H, Simpson JC, Meijer WG. (2013). A real-time impedance based method to assess Rhodococcus equi virulence. PLoS One, 8(3), e60612. https://doi.org/10.1371/journal.pone.0060612

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 8
Issue: 3
Pages: e60612
PII: e60612

Researcher Affiliations

Miranda-CasoLuengo, Aleksandra A
  • UCD School of Biomolecular and Biomedical Science and Conway Institute, University College Dublin, Dublin, Ireland.
Miranda-CasoLuengo, Raúl
    Lieggi, Nora T
      Luo, Haixia
        Simpson, Jeremy C
          Meijer, Wim G

            MeSH Terms

            • Actinomycetales Infections / microbiology
            • Actinomycetales Infections / veterinary
            • Animals
            • Cell Line
            • Electric Impedance
            • Horses / microbiology
            • Host-Pathogen Interactions
            • Humans
            • Macrophages / cytology
            • Macrophages / microbiology
            • Mice
            • Mutation
            • Rhodococcus equi / genetics
            • Rhodococcus equi / growth & development
            • Rhodococcus equi / pathogenicity
            • Rhodococcus equi / physiology

            Conflict of Interest Statement

            The authors have declared that no competing interests exist.

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            Citations

            This article has been cited 2 times.
            1. Stefańska I, Witkowski L, Rzewuska M, Dzieciątkowski T. Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens.. J Vet Med Sci 2016 May 3;78(4):543-9.
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            2. Wang X, Coulson GB, Miranda-Casoluengo AA, Miranda-Casoluengo R, Hondalus MK, Meijer WG. IcgA is a virulence factor of Rhodococcus equi that modulates intracellular growth.. Infect Immun 2014 May;82(5):1793-800.
              doi: 10.1128/IAI.01670-13pubmed: 24549327google scholar: lookup
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