The Enterococcus detection kit is based on the fast, sensitive, and proven Taqman® probe qPCR technique. The kit contains primers and a probe for detecting a highly specific sequence present on DNA of the species group Enterococcus. This kit of Sylphium molecular ecology is designed to have a sensitivity of only one molecule per reaction in environmental samples.
Enterococcus is a large genus of lactic acid bacteria of the phylum Firmicutes. Enterococci are gram-positive cocci that often occur in pairs (diplococci) or short chains, and are difficult to distinguish from streptococci on physical characteristics alone. Two species are common commensal organisms in the intestines of humans: E. faecalis (90–95%) and E. faecium (5–10%). Rare clusters of infections occur with other species, including E. casseliflavus, E. gallinarum, and E. raffinosus. (source: wikipedia)
Primers and probe and the qPCR buffer were designed to have:
- •Highest possible sensitivity (1 DNA copy per reaction). Environmental water samples contain normally very low amounts of target DNA.
- Strong fluorescence signal with low background noise. Isolated environmental samples contain residues of naturally occurring auto fluoresce substances that will interfere with the measurements. A strong fluorescence signal from the analyses is required for these kind of samples.
- 100% specificity. Isolated DNA from environmental samples contains billions of DNA fragments from bacteria, protozoa, plants, animals and must be taken in account during primer/probe design. This is validated by experimental and bioinformatical studies.
The kit is developed and optimized to be used on eDNA isolates purified using the eDNA isolation kit (#SYL002) from Sylphium molecular ecology.
Other eDNA isolation methods/kits can be used as well. Please contact us to get more information how to use the obtained isolates from other methods/kits to get reliable results with the kit.
The kit contains materials for an 8-fold analyses on 20 samples including all necessary controls.
- Positive control ( Cloned Enterococcus DNA)
- 2x Sylphium qPCR mix (100 reactions) without primers and probes
- 2x Primer/probe mix (100 reactions) for detection of Escherichia coli (FAM dye)
- 1x Taq DNA polymerase (200 reactions)
- 2 ml dilution solution (S6)
- Batch validation document