Poster Presentation Australian Society for Microbiology Annual Scientific Meeting 2016

Investigation for liaF deletion in vancomycin-resistant Enterococcus faecium isolates from Australian hospital patients (#349)

Rebecca Stephenson 1 , Steven Siarakis 1 , Genevieve McKew 1 2 , Monica Stelmach
  1. Microbiology, Concord Hospital, Sydney, NSW, Australia
  2. School of Medicine, University of Sydney, Sydney, NSW, Australia

Mutations in the gene liaF, part of the LiaFSR complex, involved in cell envelope

stress response, are known to cause daptomycin resistance (DAP-R) in Enterococcus

faecium, which can complicate the treatment of vancomycin-resistant enterococcal

infections. We recently described a case of isolation of a treatment-emergent DAP-R

VREm occurring in the context of combined daptomycin and ampicillin treatment in a

liver transplant patient. On initial WGS data analysis, a large deletion in the liaFSR

complex region was suggested. More recently, in vitro deletions in liaR, the

regulator gene of the complex, are shown to restore susceptibility to daptomycin and

telavancin in VRE E. faecalis, increase the rate of killing by human antimicrobial

peptides, and attenuate virulence in an animal model. We therefore questioned

whether Australian VRE have different genetic context under antibiotic pressure to

develop daptomycin resistance, and explored if a deletion of the LiaFSR complex was

present in Australian VRE isolates.

3 Clinical isolates, (2 being daptomycin-non- susceptible) and 94 sequential VRE

strains isolated from screening inpatients from an Australian tertiary hospital

underwent detection of liaF with real-time PCR.

Of the screening isolates, 48 of these carried the vanA gene, and 46 carried vanB.

The liaF gene was detected in 100% of the 94 isolates using RT-PCR, and detected

also in the clinical isolates.

The LiaFSR complex is found in all genrea clinically important Gram-positive bacteria,

including Streptococci, Bacillus, and Staphylococcus. It’s involved in stress response

of the cell membrane to attack by antimicrobial peptides, which have similar action

to calcium-complexed daptomycin. In vitro deletions of this complex in Enterococcus

faecalis have shown to alter daptomycin susceptibility.

On re-analysis of the WGS data from the initial isolate, alternate software was used

and liaFSR genes were normal. Assessment of a large number of sequentially stored

isolates did not find any with this deletion. It is important to confirm unusual findings

from WGS analysis using alternate methods and consider re-doing analyses using

different software.