PSTVd Detection Kits

PDD-359

PSTVd Detection Kits

Potato Spindle Tuber Viroid (PSTVd) RT-PCR Kit is designed for the detection of PSTVd specific RNA in a real-time PCR

The Potato Spindle Tuber Viroid (PSTVd) is the causal agent of a disease known as chrysanthemum stunt. The viroid consists of small pieces of RNA (354 bp) that are known to be highly contagious. The main symptoms of PSTVd infection are severe stunting of the plants such that they are half to two-thirds of the normal size, as well as smaller flowers and premature flowering. Symptoms of Chrysanthemum Stunt are difficult to detect in stock, however can be so severe at flowering that finished crops of flowers are unmarketable. There is no chemical that can control PSTVd, therefore much effort has been made to generate chrysanthemum plants free from PSTVd. Chrysanthemum Stunt Viroid SYBR Green RT-PCR Detection Kit is a rapid and sensitive PSTVd detection tool which can be applicable for the early diagnosis of PSTVd.

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 3 x 350 µL
2. PSTVd Primer & Probe Mix 3 x 70 µL
3. PSTVd Positive Control 3 x 50 µL
4. Nuclease-Free Water (Negativet l) 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)

The Potato Spindle Tuber Viroid (PSTVd) was the first viroid to be identified, and the natural hosts for this viroid are potatoes and tomatoes. Potato spindle tuber disease has been found in the United States, Canada, Russia and South Africa, and it is one of the most destructive diseases of potatoes. PSTVd is a small (359 nucleotide), circular, single-stranded RNA molecule closely related to the Chrysanthemum stunt viroid. The viroid attacks all varieties of potato, is known to spread rapidly and often occurs in combination with virus diseases. Symptoms of PSTVd infection range from mild to severe, with severe infections resulting in colour change in the foliage, smaller leaves, spindle like elongation, and a slower rate of sprouting.

PCR

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