Grapevine Red Blotch-Associated Virus (GRBaV) Pathogen Detection Kits

PDD-353

Grapevine Red Blotch-Associated Virus (GRBaV) Pathogen Detection Kits

Grapevine Red Blotch-Associated Virus (GRBaV) PCR Kit is designed for the detection of GRBaV specific DNA in a real-time PCR.

Grapevine Red Blotch-Associated Virus (GRBaV) PCR Kit is designed for the detection of GRBaV 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 GRBaV specific DNA is based on TaqMan PCR providing a simple, reliable and rapid result for the detection of GRBaV infection. Grapevine Red Blotch-Associated Virus (GRBaV) PCR Kit includes a PCR control to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The GRBaV 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.

Virus

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 350 µL
2. GRBaV Primer & Probe Mix 70 µL
3. GRBaV 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)

Grapevine red blotch-associated virus (GRBaV) is a major threat to the wine industry. GRBaV infections compromise crop yield, and cause a significant reduction in sugar, uneven ripening, or failure to ripen, resulting in off flavors and berry chemical composition, which affects the flavor and aroma properties of must and wine. Visual diagnosis is often unreliable due to several confounding abiotic and biotic factors, including similarities with leafroll disease symptoms. To date, no antibody based detection methods for GRBV are available on the market. Therefore, PCR-based assays are recommended for the accurate diagnosis of GRBV infection.

PCR

The CTiVd 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.