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Updated: Jun 8, 2025
A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters
Published on: March 9, 2016
Development of a PCR assay for rapid and accurate detection of an emerging vanB Enterococcus faecium clone in the
Maja Johanne Søndergaard Knudsen1, Christel Barker Jensen1, Rikke Lind Jørgensen1
1Department of Clinical Microbiology, Copenhagen University Hospital Hvidovre, Hvidovre, Denmark.
Objectives:
To develop and validate a real-time PCR assay detecting the sequence bridging Tn1549 and the Enterococcus faecium chromosome in the emerging vanB vancomycin-resistant E. faecium (VREfm) clone (ST80/CT2406).
Methods:
The Tn1549 insertion site was determined on routinely sequenced VREfm isolates. The outer boundaries of Tn1549 and adjoining host bacterial sequences were determined using a BLAST search in the silent information regulator gene sir2. Next, the primers and probe were developed, targeting the sequence bridging Tn1549 and the E. faecium chromosome. Finally, the PCR assay was validated on well-characterized strains and prospectively performed on rectal screening samples submitted to our laboratory.
Results And Conclusions:
The PCR assay proved to be accurate and provide rapid diagnosis of the emerging vanB VREfm in rectal screening samples.
