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Multiplex PCR for simultaneous detection of enterococcal genes vanA and vanB and staphylococcal genes mecA, ileS-2
Elena Ramos-Trujillo1, Eduardo Pérez-Roth, Sebastián Méndez-Alvarez
1Hospital Universitario Nuestra Señora de Candelaria, Unidad de Investigación, Carretera del Rosario s/n, 38010 Santa Cruz de Tenerife, Spain.
Abstract:
The experimental transfer of the vanA gene cluster from Enterococcus faecalis to Staphylococcus aureus has raised fears about the occurrence of such genetic transfer in clinical isolates of methicillin-resistant staphylococci. Recently, infections by a S. aureus strain carrying the enterococcal vancomycin resistance vanA gene cluster were reported. The possible emergence and dissemination of these strains is a serious health threat and makes optimization of prevention strategies and fast detection methods absolutely necessary. In the present study, we developed a PCR protocol for simultaneous detection of enterococcal vanA and vanB genes, the staphylococcal methicillin and mupirocin resistance markers mecA and ileS-2, and identification of S. aureus. As no vancomycin-resistant S. aureus isolates were available for our study, we used mixtures of enterococcal and staphylococcal colonies that harbored the different resistance markers to show that these genes could be detected simultaneously. This protocol could be used to facilitate the detection and identification of predictable S. aureus or methicillin-resistant strains carrying vanA or vanB.
Insights
A new PCR method can simultaneously detect vancomycin resistance genes (vanA, vanB) and identify Staphylococcus aureus, including methicillin-resistant strains. This aids in the rapid detection of dangerous, resistant bacteria.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- The transfer of vanA gene cluster from Enterococcus faecalis to Staphylococcus aureus raises concerns about clinical infections.
- Emergence of vancomycin-resistant Staphylococcus aureus (VRSA) poses a significant public health threat.
- Need for rapid detection and prevention strategies for VRSA strains.
Purpose of the Study:
- To develop a PCR protocol for simultaneous detection of vanA and vanB genes.
- To identify Staphylococcus aureus and detect methicillin (mecA) and mupirocin (ileS-2) resistance markers.
- To provide a tool for early identification of emerging resistant strains.
Main Methods:
- Development of a multiplex PCR assay.
- Simultaneous detection of enterococcal vanA and vanB genes.
- Detection of Staphylococcus aureus, mecA, and ileS-2 resistance markers.
Main Results:
- A PCR protocol was successfully developed for simultaneous gene detection.
- The protocol demonstrated the ability to detect target genes in mixed bacterial colonies.
- Validation of the method for identifying key resistance markers and bacterial species.
Conclusions:
- The developed PCR protocol facilitates the simultaneous detection of vanA/vanB genes and identification of S. aureus.
- This method can aid in the early detection of methicillin-resistant S. aureus carrying vancomycin resistance genes.
- Optimization of prevention strategies and diagnostic methods for resistant strains is crucial.
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