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Journal of clinical microbiology2003; 41(7); 3241-3245; doi: 10.1128/JCM.41.7.3241-3245.2003

Rapid identification of Rhodococcus equi by a PCR assay targeting the choE gene.

Abstract: The actinomycete Rhodococcus equi is an important pathogen of horses and an emerging opportunistic pathogen of humans. Identification of R. equi by classical bacteriological techniques is sometimes difficult, and misclassification of an isolate is not uncommon. We report here on a specific PCR assay for the rapid and reliable identification of R. equi. It is based on the amplification of a fragment of the choE gene encoding cholesterol oxidase. The choE-based PCR was assessed by using a panel of strains comprising 132 isolates from different sources and of different geographical origins, all initially identified biochemically as R. equi, and 30 isolates of representative non-R. equi actinomycete species, including cholesterol oxidase producers. The expected 959-bp amplicon was observed only with R. equi isolates, as confirmed by sequencing of a variable region of the 16S RNA gene from a random sample of 20 PCR-positive isolates. All R. equi isolates gave a positive choE-based PCR result, which correlated with a high degree of conservation of the choE gene. Three of the 132 strains originally identified as R. equi were negative for the choE gene, and subsequent analysis of their 16S RNA gene sequences confirmed that they belonged to other bacterial species (Dietzia maris, Mycobacterium peregrinum, and Staphylococcus epidermidis). All non-R. equi isolates were negative by the choE-based PCR. ATCC 21387, the only known isolate of Brevibacterium sterolicum, gave a 959-bp amplicon whose DNA sequence was virtually identical to that of R. equi choE. Comparison of the 16S RNA genes indicated that ATCC 21387 should be considered an R. equi isolate.
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Publication Date: 2003-07-05 PubMed ID: 12843070PubMed Central: PMC165360DOI: 10.1128/JCM.41.7.3241-3245.2003Google 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 article focuses on a PCR assay created to rapidly and accurately identify the bacterium Rhodococcus equi, which is a significant pathogen in horses and an emerging one in humans. The PCR test targets the choE gene related to cholesterol oxidase production and is shown to produce specific and accurate results for isolates of R. equi.

Understanding the Study and Its Methodology

  • The research article discusses the development of a PCR assay—the Polymerase Chain Reaction technique—which allows certain gene sections to be amplified for easier detection and identification. The assay targets the choE gene that encodes cholesterol oxidase, an enzyme produced by R. equi and other actinomycetes.
  • The R. equi is a problematic bacterium in horses and can cause severe diseases in humans as well. The identification of R. equi is crucial, but traditional techniques can be inaccurate leading to misclassification.
  • A panel of 132 strains initially identified as R. equi and 30 isolates of non-R. equi actinomycetes were used to assess the choE-based PCR assay. In the process, a specific DNA fragment relating to the choE gene was amplified and observed.

Findings and Results

  • From the panel, the PCR assay generated the expected 959 base pairs amplicon with R. equi isolates only. This means that the assay was successful in specifically identifying R. equi bacteria.
  • Additional testing and sequencing on 20 PCR-positive isolates confirmed the findings, highlighting the high precision of the choE-based PCR.
  • However, three out of the 132 strains originally classified as R. equi lacked the choE gene. Upon further examination of their 16S RNA gene sequences, it was determined that they belonged to different bacterial species—Dietzia maris, Mycobacterium peregrinum, and Staphylococcus epidermidis.
  • When examining the ATCC 21387 strain of Brevibacterium sterolicum, the choE-based PCR also produced a 959-bp amplicon, similar to that of R. equi. This instance suggested that ATCC 21387 could instead be considered as an R. equi isolate.

Implications

  • This choE gene-based PCR assay allows for a quick and reliable identification of R. equi. Given the health risk posed by the bacterium, particularly in horses and humans, this tool could have significant implications in medical and veterinary contexts.
  • The authors note that the high degree of conservation of the choE gene across R. equi isolates, which means this method offers robust specificity when identifying R. equi bacteria.
  • The study also underlines the potential flaws with traditional classification methods, given that three out of 132 strains were incorrectly identified as R. equi, emphasizing the need for precise diagnostic tools.

Cite This Article

APA
Ladrón N, Fernández M, Agüero J, González Zörn B, Vázquez-Boland JA, Navas J. (2003). Rapid identification of Rhodococcus equi by a PCR assay targeting the choE gene. J Clin Microbiol, 41(7), 3241-3245. https://doi.org/10.1128/JCM.41.7.3241-3245.2003

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 41
Issue: 7
Pages: 3241-3245

Researcher Affiliations

Ladrón, Néstor
  • Departamento de Biología Molecular (Unidad Asociada al Centro de Investigaciones Biológicas, C.S.I.C.), Facultad de Medicina, Universidad de Cantabria, 39011 Santander, Spain.
Fernández, Marta
    Agüero, Jesús
      González Zörn, Bruno
        Vázquez-Boland, José A
          Navas, Jesús

            MeSH Terms

            • Actinomycetales Infections / microbiology
            • Actinomycetales Infections / veterinary
            • Animals
            • Bacterial Proteins / genetics
            • Bacterial Typing Techniques
            • Brevibacterium / genetics
            • Cholesterol Oxidase / genetics
            • DNA, Ribosomal / analysis
            • Horse Diseases / microbiology
            • Horses
            • Humans
            • Molecular Sequence Data
            • Polymerase Chain Reaction / methods
            • RNA, Ribosomal, 16S / genetics
            • Rhodococcus equi / classification
            • Rhodococcus equi / enzymology
            • Rhodococcus equi / genetics
            • Rhodococcus equi / isolation & purification
            • Sensitivity and Specificity
            • Sequence Analysis, DNA
            • Time Factors

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            This article has been cited 28 times.
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