Candidatus Liberibacter solanacearum Detection Kits

PDB-167

Candidatus Liberibacter solanacearum Detection Kits

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

Candidatus Liberibacter solanacearum PCR Kit is designed for the detection of Candidatus Liberibacter solanacearum 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 solanacearum specific DNA is based on PCR providing a simple, reliable and rapid result for the detection of Candidatus Liberibacter solanacearum infection. Candidatus Liberibacter solanacearum 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 solanacearum 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 solanacearum Primer & Probe Mix 3 x 70 µL
3. Candidatus Liberibacter solanacearum Positive Control 3 x 50 µL
4. Nuclease-Free Water (Negative control) 1.25 mL 1.25 mL
5. Product Insert 1 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)

Candidatus Liberibacter solanacearum was first identified in 2008 and was shown to be associated with zebra chip disease of potato, which has been observed since the 1990s with increasing economic impacts. Candidatus Liberibacter solanacearum is also known to primarily infect tomato, pepper, eggplant, tomatillo, tamarillo, tobacco, and several weeds in the family Solanaceae. The symptoms of infection include stunting, erectness of new foliage, chlorosis and purpling of foliage with basal cupping of leaves, upward rolling of leaves throughout the plant, shortened and thickened terminal internodes resulting in plant rosetting, enlarged nodes, axillary branches or aerial tubers, leaf scorching, disruption of fruit set, and production of numerous, small, misshapen, and poor quality fruits. PCR is the method of choice for the detection of Candidatus Liberibacter solanacearum. Conventional PCR can be used, but real-time PCR is recommended because of its increased sensitivity.

PCR

The specificity of Candidatus Liberibacter solanacearum PCR Detection Kit is first and foremost ensured by the selection of the Candidatus Liberibacter solanacearum specific primers, as well as the selection of stringent reaction conditions. The Candidatus Liberibacter solanacearum 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.