Real time PCR kit for quantification of Legionella
→ Real-time PCR (qPCR) is a powerful and rapid tool for the specific detection of bacteria in water.

This methodology, widely used in most countries, is already the subject of an ISO standard for the research of Legionella spp and Legionella pneumophila in sanitary and industrial waters. It is based on the amplification and detection of specific genomic sequences. Quantification of the target DNA is performed using a specific probe labelled with the fluorophore FAM. For each amplification reaction, a synthetic DNA serves as an internal control and thus makes it possible to control the absence of inhibition and the successful running of the amplification. This internal control is quantified by a probe coupled to the HEX fluorophore. The DNA AMPLI-FLASH qPCR kit contains ready-to-use reagents for completion of the analysis.

→ The DNA AMPLI-FLASH kit, combined with the DNA PURE-FLASH kit, provides a complete, robust and reliable approach that allows the detection and quantification of Legionella in water.  

Application fields

The DNA AMPLI-FLASH kit allows an optimal detection and quantification of Legionella spp and Legionella pneumophila. The analysis is performed after extraction and purification (DNA PURE-FLASH kit) from both SANITARY WATER samples (drinking water, hot and cold water network, hydrotherapy, swimming pools…) and INDUSTRIAL WATER samples (cooling installations, washing stations…).

Protocol key points

1. Preparation of the mix, combine the Master Mix solution with the Prim’n’Prob solution.

2. Distribute 15 µl of mix to each PCR wells, according to the classical protocol.

3. Add 5 µl of DNA extract per PCR well.

4. Start the PCR run using the appropriate thermoprofile.

Advantages of
DNA
AMPLI-FLASH
 technology

RAPID

results obtained in under 2 hours

SIMPLE

Handling of 2 reagents and pipetting table

PERFORMANCE

Validated according to NF T90-471 and ISO 12869, applicable on many different samples

EFFECTIVE

A single thermo-profile for both PCR systems (Legionella spp and pneumophila)

FLEXIBLE

Compatible with many different thermal cyclers (Chromo IV and CFX96 – Bio-rad, MIC - Bio molecular systems, 7500/7500 fast – Applied)

LIMIT OF DETECTION

5 GU
per PCR well (5µl)

LIMIT OF QUANTIFICATION

25 GU
per PCR well (5µl)

1
THERMOPROFILE
for both PCR systems

Up to

25 000 GU
quantifiable per well

Kit content for 96 reactions

– 1 tube of Master Mix,
– 1 tube of « Prim’n’Prob »,
– 1 tube of « Negative control »,
– 1 tube of « Positive control »,
– 1 tube of calibrated solution of Legionella « Standard solution »,
– 1 tube of « Dilution buffer ».

Equipment required

– Pipettes P10, P100, P1000 and precision tips,
– Microtubes of 1.5 and 2.0 ml,
– PCR tubes or plate,
– Thermocycler,
– Vortex.

We also recommend:

MIC qPCR cycler

For quantification of Legionella by qPCR.

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