mericon Quant L. pneumophila Kit
For real-time PCR detection and quantification of Legionella pneumophila in water
The mericon Quant L. pneumophila Kit is designed for the target-specific detection and quantification of Legionella pneumophila DNA in water after concentration (e.g., by filtration). The assay uses a real-time PCR-based protocol and forms part of the comprehensive QIAGEN food testing portfolio, which also features dedicated sample preparation kits and assays for ingredient authentication and GMO DNA detection. The mericon Quant L. pneumophila assay performs optimally on the Rotor-Gene Q, but has also been validated for block thermal cyclers.
The mericon Quant L. pneumophila Kit is intended for molecular biology applications in food, water, animal feed, and pharmaceutical product testing. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance
The mericon Quant L. pneumophila Kit is for the specific detection and quantification of Legionella pneumophila in water after concentration (e.g., by filtration). The assay can detect as few as 10 copies of L. pneumophila DNA in a reaction and it exhibits high specificity for L. pneumophila . In an experiment using 2500 copies of the tested DNA, no cross-reactivity was observed with other pathogens (see the table "Results of mericon Quant L. pneumophila Kit cross-reactivity study"). Results of mericon Quant L. pneumophila Kit cross-reactivity study
Principle
The mericon Quant L. pneumophila Kit is designed for the quantitative detection of Legionella pneumophila in water after filtration. L. pneumophila is the Legionella species most frequently involved in human infections, most notably in the so-called Legionnaires' disease. Detection of pathogens using real-time PCR is based on the amplification of a specific region of the relevant pathogen genome. The amplified product is detected using target-specific fluorescent probes that bind to it. As the PCR product accumulates, there is an increased fluorescent signal from the bound probes. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating PCR product without having to re-open the reaction tubes afterward. Quantification of the target Legionella species is carried out based on a standard curve covering a concentration range of 25,000–25 copies per PCR reaction. The standard curve is prepared from purified Legionella Standard DNA, which is provided in the mericon Quant L. pneumophila Kit at an accurately defined concentration. mericon PCR Assays include an internal control to control potential inhibition. This aids in the straightforward interpretation of the results (see the table "Summary of possible PCR outcomes"). The PCR primer set for each assay is highly specific and targets a unique and conserved region of the tested pathogen genome that has been bioinformatically and experimentally verified.
Procedure
DNA is extracted from a filtered or concentrated water sample using a dedicated sample preparation kit. The isolated, pure bacterial DNA is assayed using a streamlined real-time PCR protocol that returns results in 73 minutes on QIAGEN's Rotor-Gene Q. The protocol has also been validated for use on other thermal block cyclers. The results are displayed as easy-to-interpret PCR curves.
Applications
The mericon Quant L. pneumophila Kit is for real-time PCR-based detection and quantification of L. pneumophila in water.
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