CEVd Detection Kits

PDD-350

CEVd Detection Kits

Citrus Exocortis Viroid (CEVd) RT-PCR Kit is designed for the detection of CEVd specific RNA in a real-time PCR.

Citrus Exocortis Viroid (CEVd) RT-PCR Kit is designed for the detection of CEVd specific RNA 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 CEVd specific RNA is based on one-step RT-PCR providing a simple, reliable and rapid result for the detection of CEVd infection. CEVd RT-PCR Kit includes a PCR control to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The CEVd RT-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.

Viroid

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 RT-PCR Master Mix 350 µL
2. CEVd Primer & Probe Mix 70 µL
3. CEVd Positive Control 50 µL
4. Nuclease-Free Water (Negative control) 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 Exocortis Viroid (CEVd) is a member of the genus Pospiviroid within the family Pospiviroidae. CEVd is the causal agent of the exocortis disease characterized by a bark shelling or scaling disorder of trifoliate orange used as rootstock. The citrange hybrids and Rangpur lime, both used as rootstocks, are also sensitive and develop bark scaling symptoms and stunting. Viroids are small plant pathogens consisting of a naked single-stranded circular RNA molecule of 246–475 nt, that do not encode any proteins but are endowed with autonomous replication in their host plants. Citrus Exocortis Viroid RT-PCR Detection Kit is a rapid and sensitive CEVd detection tool which can be applicable for the early diagnosis of CEVd.

PCR

The specificity of CEVd TaqMan RT-PCR Kit is first and foremost ensured by the selection of the CEVd-specific primers, as well as the selection of stringent reaction conditions. The CEVd primers were checked for possible homologies to all plant viroids 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.