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Home / Equine Research Bank / Animals (Basel) / Novel Quantitative PCR for Rhodococcus equi and Macrolide Re...
Animals : an open access journal from MDPI2022; 12(9); 1172; doi: 10.3390/ani12091172

Novel Quantitative PCR for Rhodococcus equi and Macrolide Resistance Detection in Equine Respiratory Samples.

Abstract: is an important veterinary pathogen that takes the lives of many foals every year. With the emergence and spread of MDR to current antimicrobial treatment, new tools that can provide a fast and accurate diagnosis of the disease and antimicrobial resistance profile are needed. Here, we have developed and analytically validated a multiplex qPCR for the simultaneous detection of and related macrolide resistance genes in equine respiratory samples. The three sets of oligos designed in this study to identify housekeeping gene and macrolide resistance genes (46) and (51) showed high analytic sensitivity with a limit of detection (LOD) individually and in combination below 12 complete genome copies per PCR reaction, and an amplification efficiency between 90% and 147%. Additionally, our multiplex qPCR shows high specificity in in-silico analysis. Furthermore, it did not present any cross-reaction with normal flora from the equine respiratory tract, nor commonly encountered respiratory pathogens in horses or other genetically close organisms. Our new quantitative PCR is a trustable tool that will improve the speed of infection diagnosis, as well as helping in treatment selection.
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Publication Date: 2022-05-03 PubMed ID: 35565598PubMed Central: PMC9099730DOI: 10.3390/ani12091172Google 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 researchers of this study developed a multiplex qPCR technique that simultaneously detects Rhodococcus equi, a deadly pathogen in foals, and its associated macrolide resistance genes in equine respiratory samples. The new tool provides quick and precise diagnosis of the infection and antibiotic resistance profile.

Development of the Multiplex qPCR Technique

  • The study was motivated by the need for a fast and accurate diagnostic and prognosis tool for the Rhodococcus equi infection in foals and the ability to profile its antibiotic resistance. R. equi has been increasingly demonstrating Multi-Drug Resistance (MDR) to existing antimicrobial treatments, adversely affecting efficiency in controlling the infection.
  • The multiplex quantitative PCR (qPCR) allows for the simultaneous detection of R. equi and associated resistance genes to macrolide antibiotics. This provides real-time diagnosis and resistance profiling, thereby improving disease control measures and treatment outcomes.

Validation of the Multiplex qPCR Technique

  • The analytical validation of the tool involved designing three sets of oligonucleotides. These sequences target the housekeeping gene of R. equi and two specific resistance genes (46 and 51) related to macrolide antibiotics.
  • The diagnostic tool demonstrated high analytic sensitivity, with the limit of detection (LOD) for each target and in combination falling below 12 complete genome copies per PCR reaction. These positive results indicate the tool’s reliability in detecting minimal traces of R. equi and its resistance genes in test samples.
  • Additionally, the technique had an amplification efficiency ranging between 90% and 147%, highlighting its high yield in successfully identifying targeted sequences in samples.
  • The multiplex qPCR also demonstrated high specificity in in-silico (computer simulated) analysis, proving its effectiveness in accurately identifying and differentiating target genes from non-targets.
  • The developed technique showed no cross-reaction with the normal flora of the equine respiratory tract or commonly encountered respiratory pathogens in horses or other genetically similar organisms. This lack of cross-reaction further indicates the high specificity and reliability of the new technique.

Advantages of the Multiplex qPCR Technique

  • The new qPCR tool offers a faster method for detecting R. equi infection. This quick diagnosis will allow for timely and accurate treatment, thus improving the prognosis of affected foals.
  • Additionally, by profiling the resistance genes alongside the pathogen, the tool informs the most suitable choice of antibiotic treatment. This information will help overcome the challenge of MDR and improve the effectiveness of treatments against R. equi.

Cite This Article

APA
Narváez SÁ, Fernández I, Patel NV, Sánchez S. (2022). Novel Quantitative PCR for Rhodococcus equi and Macrolide Resistance Detection in Equine Respiratory Samples. Animals (Basel), 12(9), 1172. https://doi.org/10.3390/ani12091172

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 12
Issue: 9
PII: 1172

Researcher Affiliations

Narváez, Sonsiray Álvarez
  • Poultry Diagnostic and Research Center, Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.
Fernández, Ingrid
  • Athens Veterinary Diagnostic Laboratory, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.
Patel, Nikita V
  • Athens Veterinary Diagnostic Laboratory, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.
Sánchez, Susan
  • Athens Veterinary Diagnostic Laboratory, Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.

Grant Funding

  • U18 FD004623 / FDA HHS
  • 1U18FD006157-04 / United States Food and Drug Administration

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 2 times.
  1. Ghielmetti G, Stevens MJA, Schmitt S, Kittl S, Cernela N, Biggel M, Schulthess B, Keller PM, Schrenzel J, Stephan R. Multi-host distribution of Rhodococcus equi (Prescottella equi) strains and their phylogenomic clustering. BMC Microbiol 2025 Jul 21;25(1):447.
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  2. Vandersnickt J, Van PJ, Vandamme S, Kenyon C, Florence E, Van Ierssel S, Vlieghe E, Theunissen C. Rhodococcus equi, an Unusual Human Pathogen That Causes Cavitating Pneumonia in Patients With AIDS. Case Rep Infect Dis 2024;2024:5570186.
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