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Volume 14
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10.3390/antibiotics14030295
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Article

Multiplex Real-Time Polymerase Chain Reaction and Recombinase Polymerase Amplification: Methods for Quick and Cost-Effective Detection of Vancomycin-Resistant Enterococci (VRE)

by
Ibukun Elizabeth Osadare
1,2,
Abdinasir Abdilahi
1,2,
Martin Reinicke
1,2,
Celia Diezel
1,2,
Maximilian Collatz
1,2,
Annett Reissig
1,2,
Stefan Monecke
1,2 and
Ralf Ehricht
1,2,3,*
1
Leibniz Institute of Photonic Technology (IPHT), Member Research Alliance Leibniz Centre for Photonics in Infection Research (LPI), 07745 Jena, Germany
2
InfectoGnostics Research Campus, 07743 Jena, Germany
3
Institute of Physical Chemistry, Friedrich-Schiller University, 07743 Jena, Germany
*
Author to whom correspondence should be addressed.
Antibiotics 2025, 14(3), 295; https://doi.org/10.3390/antibiotics14030295
Submission received: 31 January 2025 / Revised: 3 March 2025 / Accepted: 10 March 2025 / Published: 12 March 2025
(This article belongs to the Special Issue Genomic Insights and Implications of Emerging Antimicrobial Resistance)
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Abstract

Background/Objectives: Vancomycin-resistant enterococci (VRE) are one of the leading causes of antibiotic-resistant infections in the hospital setting worldwide, and this has become a major issue, because most patients infected with this strain are difficult to treat. Multiplex real-time polymerase chain reaction (RT PCR) is an advantageous technique that can amplify multiple targets in a single reaction, and can be used to quickly detect specific targets in VRE within two hours, starting from suspected colonies of bacterial cultures, without sample preparation. Methods: In this study, we selected the glycopeptide/vancomycin resistance genes that are most common in clinical settings, vanA and vanB, in combination with the species markers ddl_faecium and ddl_faecalis for the most common VRE species—Enterococcus faecium and Enterococcus faecalis. Results: DNA from forty clinical VRE strains was prepared using a fast and economic heat lysis method, and a multiplex real-time PCR assay was optimized and carried out subsequently. The results were in concordance with the results from recombinase polymerase amplification (RPA) of the same VRE samples. Conclusions: Multiplex RT PCR and RPA for VRE detection proffers a second method for the confirmation of vancomycin resistance, and it can be developed as a fast screening assay for patients before admission into high-risk settings.
Keywords: Enterococcus spp.; VRE; resistance genes; DNA; PCR; RPA Enterococcus spp.; VRE; resistance genes; DNA; PCR; RPA

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MDPI and ACS Style

Osadare, I.E.; Abdilahi, A.; Reinicke, M.; Diezel, C.; Collatz, M.; Reissig, A.; Monecke, S.; Ehricht, R. Multiplex Real-Time Polymerase Chain Reaction and Recombinase Polymerase Amplification: Methods for Quick and Cost-Effective Detection of Vancomycin-Resistant Enterococci (VRE). Antibiotics 2025, 14, 295. https://doi.org/10.3390/antibiotics14030295

AMA Style

Osadare IE, Abdilahi A, Reinicke M, Diezel C, Collatz M, Reissig A, Monecke S, Ehricht R. Multiplex Real-Time Polymerase Chain Reaction and Recombinase Polymerase Amplification: Methods for Quick and Cost-Effective Detection of Vancomycin-Resistant Enterococci (VRE). Antibiotics. 2025; 14(3):295. https://doi.org/10.3390/antibiotics14030295

Chicago/Turabian Style

Osadare, Ibukun Elizabeth, Abdinasir Abdilahi, Martin Reinicke, Celia Diezel, Maximilian Collatz, Annett Reissig, Stefan Monecke, and Ralf Ehricht. 2025. "Multiplex Real-Time Polymerase Chain Reaction and Recombinase Polymerase Amplification: Methods for Quick and Cost-Effective Detection of Vancomycin-Resistant Enterococci (VRE)" Antibiotics 14, no. 3: 295. https://doi.org/10.3390/antibiotics14030295

APA Style

Osadare, I. E., Abdilahi, A., Reinicke, M., Diezel, C., Collatz, M., Reissig, A., Monecke, S., & Ehricht, R. (2025). Multiplex Real-Time Polymerase Chain Reaction and Recombinase Polymerase Amplification: Methods for Quick and Cost-Effective Detection of Vancomycin-Resistant Enterococci (VRE). Antibiotics, 14(3), 295. https://doi.org/10.3390/antibiotics14030295

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