PCRmax Enterococcus faecium Instruction Manual

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Enterococcus faecium

PCRmax Ltd qPCR test

TM

150 tests

For general laboratory and research use only

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E. faecium is a Gram-positive, non-motile bacterium of spherical shape that occurs in pairs
or chains. The DNA genome of approximately 2.8 Mbp has a circular organisation with
several plasmids. These plasmids are thought to play a role in antibiotic resistance by
conjugation. This bacterium is a facultative anaerobe that usually inhabits the
gastrointestinal tract of many animal species. However, it can colonise the urinary tract or
open wounds causing infection. Strains of E. faecium found in clinical isolates are often
found to possess enterococcal surface proteins (Esp) that participate in bacterial
aggregation and the formation of biofilms.

The bacteria can survive for long periods of time on many surfaces and in environments
including soil. E. faecium can also withstand temperatures of 10-45°C and acidic or basic
environments. From any surface the bacteria can be transferred to skin where it infects any
open wound. E. faecium has a highly significant role in nosocomial infections resulting in
bacteremia when hospital surfaces are not thoroughly sterilised. The expression of Esp
gene aids this by increasing the ability to bind to hospital instruments in the bio-film
configuration. Conjugation of plasmids has lead to multi-drug resistance including
resistance to the antibiotic Vancomycin.

Infection by E. faecium can lead to nosocomial bacteremia, surgical wound infection,
endocarditis and urinary tract infections depending on how the patient was initially infected.

Introduction to Enterococcus faecium

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MIN

MAX

The primers have 100% homology with all Enterococcus faecium reference sequences
included in the NCBI database (AF417582.1 and AY315820.1) and have low homology to
other Enterococcus, Lactococcus and Streptococcus strains.

The PCR Max qPCR Kit for Enterococcus faecium (E.faecium) genomes is designed for
the in vitro quantification of E.faecium genomes. The kit is designed to have the broadest
detection profile possible whilst remaining specific to the E.faecium genome.

The primers and probe sequences in this kit have 100% homology with a broad range of
E.faecium sequences based on a comprehensive bioinformatics analysis.

If you require further information, or have a specific question about the detection profile of
this kit then please send an e.mail to [e-mail] and our bioinformatics team will
answer your question.

Specificity

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Kit Contents

• E.faecium specific primer/probe mix (150 reactions BROWN)

FAM labelled

• E.faecium positive control template (for Standard curve RED)

• Internal extraction control primer/probe mix (150 reactions BROWN)

VIC labelled as standard

• Internal extraction control DNA (150 reactions BLUE)

• Endogenous control primer/probe mix (150 reactions BROWN)

FAM labelled

• RNAse/DNAse free water (WHITE)

for resuspension of primer/probe mixes and internal extraction control DNA

• Template preparation buffer (YELLOW)

for resuspension of positive control template and standard curve preparation

Reagents and equipment to be supplied by the user

Real-Time PCR Instrument

DNA extraction kit

This kit designed to work well with all processes that yield high quality DNA with minimal
PCR inhibitors.

Lyophilised 2x qPCR Mastermix

This kit is designed to be compatible with all commercially available Mastermixes that run
with standard cycling conditions.

Pipettors and Tips

Vortex and centrifuge

Thin walled 1.5 ml PCR reaction tubes

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