Pospiviroid Detection Kits

PDD-358

Pospiviroid Detection Kits

Pospiviroid RT-PCR Kit is designed for the detection of Pospiviroid specific RNA in a real-time PCR.

Pospiviroid RT-PCR Kit is designed for the detection of Pospiviroid 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 Pospiviroid specific RNA is based on one-step RT-PCR providing a simple, reliable and rapid result for the detection of Pospiviroid infection. Pospiviroid 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 Pospiviroid 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. Pospiviroid Primer & Probe Mix 70 µL
3. Pospiviroid 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)

Pospiviroid belongs to the Pospiviroidae family on the basis of biochemical, biological and structural properties which have been classified. The genus Pospiviroid contains nine viroid species: Potato spindle tuber viroid (PSTVd), Tomato chlorotic dwarf viroid (TCDVd), Mexican papito viroid (MPVd), Tomato planta macho viroid (TPMVd), Chrysanthemum stunt viroid (CSVd), Citrus exocortis viroid (CEVd), Tomato apical stunt viroid (TASVd), Iresine viroid (IrVd) and Columnea latent viroid (CLVd). Viroids cannot be detected by immunological methods because they do not encode any proteins, therefore an accurate and reliable diagnostic method for the detection of viroids, followed by eradication of the infected sources, is needed for quarantine operations. Pospiviroid Viroid RT-PCR Detection Kit allows for the detection of any of the 9 viroid species of the Pospiviroid family.

PCR

The specificity of Pospiviroid RT-PCR Kit is first and foremost ensured by the selection of the Pospiviroid-specific primers, as well as the selection of stringent reaction conditions. The Pospiviroid 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.