Legionella sp. Detection Kits

PDB-169

Legionella sp. Detection Kits

Legionella sp. PCR Kit Dx is an in vitro diagnostic test for the detection and quantification of Legionella sp. specific DNA using real-time hybridization-fluorescence detection.

Legionella sp.PCR Kit Dx is an in vitro diagnostic test for the detection and quantification of Legionella sp. specific DNA using real-time hybridization-fluorescence detection. The kit includes a PCR control to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The Legionella sp. PCR Kit Dx comprises Master Mix for the target and PCR control detection, Primer & Probe Mix, as well as a positive control and a negative control (nuclease-free water) to confirm the integrity of the kit reagents. Legionella sp. PCR Kit Dx is based on real-time PCR technology, utilizing PCR for the amplification of specific target sequences and target specific probes for the detection of the amplified DNA. The probes are labelled with fluorescent reporter and quencher dyes. Probes specific for Legionella sp. are labelled with the fluorophore FAM. The probe specific for the PCR control is labelled with the fluorophore HEX. Using probes linked to distinguishable dyes enables the parallel detection of Legionella sp. specific DNA and the PCR control in corresponding detector channels of the real-time PCR instrument.

1. Before use, suitable amounts of all components should be completely thawed at room temperature, mixed by gentle vortexing or by pipetting, and centrifuged briefly.
2. Work quickly on ice.
3. The recommended minimum number of DNA samples tested per PCR run is 6.

Bacteria

1. Follow universal precautions. All specimens should be considered as potentially infectious and handled accordingly.
2. Ensure that a suitable lab coat, disposable gloves, and protective goggles are worn when handling specimens and kit reagents.
3. Use sterile pipette tips with filters. Use proper pipetting techniques and maintain the same pipetting pattern throughout the procedure to ensure optimal and reproducible values.
4. Ensure that appropriate specimen collection, transport, storage, and processing techniques are followed for optimal performance of this test.

1. MDx 2X PCR Master Mix Dx 550 µL
2. Legionella sp. Primer & Probe Mix Dx 2 x 70 µL
3. Legionella sp. Positive Control Dx –200,000 copies/μL 50 µL
4. Nuclease-Free Water 2 x 1.25 mL
5. Product Insert 1

1. Appropriate Real-Time PCR Instrument with FAM and HEX filter channel
2. DNA Purification Kit
3. Disposable powder-free gloves
4. Benchtop microcentrifuge
5. Micropipettors
6. Sterile pipette tips with filters
7. PCR tubes
8. Vortex mixer
9. PCR reaction preparation station (Optional)

Legionnaires’ disease (LD) is an often severe and potentially fatal form of bacterial pneumonia caused by inhalation of aerosols generated from water or environmental sources contaminated with Legionella sp. These ubiquitous freshwater and soil inhabitants cause human respiratory disease when amplified in manmade water or cooling systems and their aerosols expose a susceptible population. The organism often responds to antimicrobial therapy, usually with macrolides, and clinical responses usually occur within 3 to 5 days. Pulmonary infections caused by Legionella may be subclinical or severe, and the fatality rate can approach 50% in immunocompromised patients.

Real-time hybridization-fluorescence detection

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots. All reagents can be stored for 1 year at -20°C without showing any reduction in performance.