Related Experiment Videos
A PCR assay for rapid detection of vancomycin-resistant enterococci
X Pérez-Hernández1, S Méndez-Alvarez, F Claverie-Martín
1Molecular Biology Laboratory, Research Unit, Nuestra Señora de Candelaria University Hospital, Santa Cruz de Tenerife, Spain.
Abstract:
Since the first report of a vancomycin-resistant enterococcal clinical isolate, these Gram-positive bacteria have emerged as important nosocomial pathogens. Several glycopeptide resistance phenotypes can be distinguished on the basis of the level and inducibility of resistance to vancomycin and teicoplanin. In the present study, we developed a multiplex PCR, which allows the simultaneous identification of enterococci at the genus level and detection of the most frequent glycopeptide resistance genotypes. Five primer sets targeting the genes vanA, vanB, vanC1, vanC2/C3 and tuf were used in one reaction tube with bacterial DNA extracted from three to five colonies. This PCR method is suitable for the rapid detection of vancomycin-resistant enterococci.
Insights
A new multiplex PCR method rapidly identifies vancomycin-resistant enterococci (VRE) and their resistance genes. This tool aids in quickly detecting these significant nosocomial pathogens.
Area of Science:
- Microbiology
- Molecular Biology
- Clinical Diagnostics
Background:
- Vancomycin-resistant enterococci (VRE) are significant nosocomial pathogens.
- Distinct glycopeptide resistance phenotypes exist, influencing treatment strategies.
Purpose of the Study:
- To develop a rapid and simultaneous method for identifying enterococci and detecting common glycopeptide resistance genes.
- To provide a tool for quick clinical detection of VRE.
Main Methods:
- Development of a multiplex PCR assay.
- Utilized five primer sets targeting vanA, vanB, vanC1, vanC2/C3, and tuf genes.
- Tested with bacterial DNA from enterococcal colonies.
Main Results:
- The multiplex PCR successfully identified enterococci at the genus level.
- Simultaneously detected the most frequent glycopeptide resistance genotypes.
- Demonstrated suitability for rapid VRE detection.
Conclusions:
- The developed multiplex PCR is an effective method for the rapid identification of VRE.
- This assay facilitates timely clinical decisions regarding VRE infections.