Candidatus Liberibacter Detection Kits

PDB-166

Candidatus Liberibacter Detection Kits

Candidatus Liberibacter PCR Kit is designed for the detection of Candidatus Liberibacter specific DNA in a real-time PCR.

Candidatus Liberibacter PCR Kit is designed for the detection of Candidatus Liberibacter specific DNA in a real-time PCR based on the use of technology. This kit is designed for research use only and not for use in diagnostic procedures. The detection of Candidatus Liberibacter specific DNA is based on PCR providing a simple, reliable and rapid result for the detection of Candidatus Liberibacter infection. Candidatus Liberibacter PCR Kit includes a PCR control to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The Candidatus Liberibacter PCR Kit 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.

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 3 x 350 µL
2. Candidatus Liberibacter Primer & Probe Mix 3 x 70 µL
3. Candidatus Liberibacter Positive Control 50 µL 3 x 50 µL
4. Nuclease-Free Water (Negative control) 1.25 mL 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)

Citrus Huanglongbing (HLB), previously known as citrus greening, constitutes the most economically devastating disease of citrus worldwide, being present in around 40 countries. HLB causes severe symptoms, depending on the age and the number of infections per plant. The disease is caused by the fastidious gram-negative α-proteobacteria Candidatus Liberibacter asiaticus (Las), Ca. L. africanus, and Ca. L. americanus as well as some phytoplasmas. The bacterium propagates within the phloem of citrus plants producing die-back, yellow shoots, blotchy mottles on leaves, and off-tasting and malformed fruit. The ‘Ca. Liberibacter’ bacterium is transmitted by the psyllids Trioza erytreae in Africa and Diaphorina citri in Asia and the Americas. A variety of diagnostic methods have been developed to supplement or confirm field-based diagnosis of HLB however, end-point PCR-based and real-time quantitative methods are currently the most widely used methods in plant and insect tissues to detect and identify HLB- Liberibacters.

PCR

The specificity of Candidatus Liberibacter PCR Detection Kit is first and foremost ensured by the selection of the Candidatus Liberibacter specific primers, as well as the selection of stringent reaction conditions. The Candidatus Liberibacter primers were checked for possible homologies to all microorganisms in GenBank published sequences by sequence comparison analysis.

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.