HSVd Detection Kits

PDD-355

HSVd Detection Kits

HSVd RT-PCR Kit is designed for the detection of Hop Stunt Viroid specific RNA in a real-time PCR.

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

The Hop Stunt Viroid (HSVd) is one of a group of the smallest self-replicating RNAs pathogenic to plants. The viroid is a covalently closed, circular, single-stranded RNA consisting of 297 nucleotides. Strains of HSVd can be found to infect citrus, grapes, stone fruit and cucumber, and have been detected in Europe, Asia, North America, Australia and the Middle East. To prevent or reduce the negative effects caused by the viroid it is necessary to use viroid-free planting material. Methods for the identification of this viroid in host plants should help to control the spread of this agriculturally important plant pathogen.

PCR

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