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The Journal of veterinary medical science2015; 78(4); 543-549; doi: 10.1292/jvms.15-0516

Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens.

Abstract: Rhodococcus equi is the causative agent of rhodococcosis in horses, resulting in significant morbidity and mortality in foals. This bacterium has also been isolated from a variety of animals and is being increasingly reported as a cause of infection in humans, mainly in immunosuppressed individuals. Laboratory diagnostics of R. equi infections based only on conventional microbiological methods shows low accuracy and can lead to misidentification. The objective of the study was to develop and evaluate a real-time PCR assay for direct detection of R. equi in various clinical specimens, including tissue samples. The species-specific region of the gene encoding R. equi cholesterol oxidase, choE, was used as a qPCR-target. The diagnostic applicability of the assay was confirmed by testing various tissue specimens obtained from horses with clinical signs of rhodoccocal infection and swine submaxillary lymph nodes. The rate of R. equi detection in clinical specimens by the developed assay was higher in comparison to the culture method (90% vs. 60.0% of positive samples) and conventional PCR (90.0% vs. 20.0% of positive samples). In case of 13 samples that were negative in the culture-based method, R. equi was detected by the developed assay. Only in one case, it gave negative result for culture-positive sample. The assay may provide a simple and rapid tool to complement the classical methods of R. equi detection based on culture and phenotypic identification of isolates, as the performed evaluation indicated a high specificity and accuracy of the results.
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Publication Date: 2015-12-08 PubMed ID: 26655770PubMed Central: PMC4873842DOI: 10.1292/jvms.15-0516Google Scholar: Lookup
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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.

The research focuses on the development and evaluation of a real-time PCR assay for the detection of Rhodococcus equi – a bacterium that causes infection in a variety of animals and worsening in immunosuppressed humans – in various clinical specimens.

Objective of the Research

  • The core objective of this study was to develop and evaluate a real-time Polymerase Chain Reaction (PCR) assay that directly detects Rhodococcus equi in different clinical specimens, especially in tissue samples.

The Development and Testing Methodology

  • The unique region of the gene responsible for R. equi cholesterol oxidase, choE, was used as the PCR target.
  • The assay’s diagnostic applicability was certified following a test on various tissue samples obtained from horses that displayed signs of rhodococcal infection and swine submaxillary lymph nodes.

Results from the Application of the Developed Assay

  • Comparison of the assay with conventional culture and PCR methods for detecting R. equi showed that the developed method was more effective. Specifically, the detection rate with the new assay hit 90% against 60% and 20% for culture method and conventional PCR method respectively.
  • Among 13 samples that scored negative with the culture-based detection, the newly developed assay positively identified R. equi.
  • Only in a single case the assay gave a negative result for a previously confirmed positive culture sample.

Conclusion

  • The study concludes that the new, real-time PCR assay offers a quick and straightforward tool to complement traditional methods of R. equi detection, which are centred on culture and phenotypic identification of isolates.
  • The results from the assessment showed that the assay is specific and accurate in R. equi detection, outperforming culture-based and conventional PCR methods. Therefore, it may be considered for routine detection of this bacterium as it improves the accuracy of diagnosis and subsequently the efficiency of treatment.

Cite This Article

APA
Stefańska I, Witkowski L, Rzewuska M, Dzieciątkowski T. (2015). Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens. J Vet Med Sci, 78(4), 543-549. https://doi.org/10.1292/jvms.15-0516

Publication

The Journal of veterinary medical science
ISSN: 1347-7439
NlmUniqueID: 9105360
Country: Japan
Language: English
Volume: 78
Issue: 4
Pages: 543-549

Researcher Affiliations

Stefańska, Ilona
  • Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, 8 Ciszewskiego St., 02-786 Warsaw, Poland.
Witkowski, Lucjan
    Rzewuska, Magdalena
      Dzieciątkowski, Tomasz

        MeSH Terms

        • Actinomycetales Infections / diagnosis
        • Actinomycetales Infections / microbiology
        • Actinomycetales Infections / veterinary
        • Animals
        • Cholesterol Oxidase / analysis
        • Cholesterol Oxidase / genetics
        • Horse Diseases / diagnosis
        • Horse Diseases / microbiology
        • Horses
        • Real-Time Polymerase Chain Reaction / veterinary
        • Rhodococcus equi / isolation & purification
        • Swine
        • Swine Diseases / diagnosis
        • Swine Diseases / microbiology

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        Citations

        This article has been cited 1 times.
        1. de Alegría Puig CR, Pilares L, Marco F, Vila J, Martínez-Martínez L, Navas J. Comparison of the Vitek MS and Bruker Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Systems for Identification of Rhodococcus equi and Dietzia spp. J Clin Microbiol 2017 Jul;55(7):2255-2260.
          doi: 10.1128/JCM.00377-17pubmed: 28490491google scholar: lookup
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