Listeria Detection Kit

MAMBA® orange line – Identification of Microorganism

This qPCR Listeria monocytogenes with 139E variant detection kit targets the zinc metalloproteinase (mpl) gene from Listeria monocytogenes.

Listeria monocytogenes causes listeriosis. With Listeria contaminated food including milk and different kind of meats are the main sources of infections from Listeria. Therefore detection of this bacteria is important for animal and human health by testing food and feed products. To provide a meaningful test we provide an internal control to minimize false results. This kit detects Listeria monocytogenes via a specific probe which can be measured in the FAM channel after amplification. Lately a new Variant (139E) was shown to not be recognized by many kits due to differences in the mastermix. With this kit the classical Listeria monocytogenes as well as the variant can be reliable detected. It provides anything for the Real-Time-PCR as well as analysis and interpretation. The internal control is detected via hex channel. We advise to use the protocol DIN EN ISO 11290-1 for the cultivation and enrichment as well as the MAMBA® grey line Listeria monocytogenes DNA Extraction kit.  

The kit is designed for 96 reactions with a final reaction volume of 20 µl including at least one positive and one negative control.

  • Detects all Listeria species and serotypes
  • Additionally detects the V139E variant
  • REF: OR-002-96x
Data sheet
Workflow
Manual

Applicability

  • Compatible with lowand high-throughput qPCR instruments.
  • Validated with BioRad CFX ConnectTM, Applied Biosystem 7500 FAST, QuantStudioTM
  • Utilizes conventional FAM and HEX marked probes.
  • Multiplex approach. Each well contains an internal control to detect sample inhibition.
  • Shelf life: 1 year at −20 °C or 6 months at 4 °C.
  • Includes 5 freeze and thaw cycles
  • Two pipetting steps required to prepare the ready-to-use PCR mix, the risk of contamination is significantly reduced.
  • Results in less than 1.5 hours

Specificity

Detects all Listeria species and serotypes, especially Variant 139E.
Tested against a various number of closely related bacteria and DNAs relevant in animal feed.

Exclusivity

Target DNA at low levels (≈5 copies)
was successfully detected in mixed
samples containing high levels of
non-target bacterial DNA (≈ 5000
copies).

Sensitivity

Capable of detecting up to 5 copies of Listeria plasmid DNA, even in the presence of a high background flora. 

Multiple copy numbers tested: 1000 (left), 100 (middle), 5 (right)

  • red = Listeria monocytogenes
  • blue = internal control
Sensitivity results

Multiple copy numbers tested: 1000 (left), 100 (middle), 5 (right)

  • green = Listeria monocytogenes variant
  • blue = internal control

Reliability

System remains stable and robust in relation to temperature (±1 °C), mastermix volume (-6 %), probe and primer concentration (-30 %) variations

Reliability results

Tested under standard (60 °C) and triple altered
conditions (59 °C).

  • red = Listeria mono. with variant 139E
  • blue = internal control