PPV Detection Kits

PDD-357

PPV Detection Kits

PPV RT-PCR Kit is designed for the detection of Plum Pox Virus specific RNA in a real-time PCR.

PPV RT-PCR Kit is designed for the detection of PPV-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 PPV-specific RNA is based on one-step RT-PCR providing a simple, reliable, and rapid result for the detection of PPV infection. PPV 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 PPV RT-PCR Kit comprises a Master Mix for the target and PCR control detection, Primer & Probe Mix, as well as a positive control and 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 RT-PCR Master Mix 350 µL
2. PPV Primer & Probe Mix 70 µL
3. PPV 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)

Plum pox virus (PPV) is a member of the genus Potyvirus, and the virus is considered the most serious pathogen affecting stone fruits including apricots, cherry, nectarine, peaches, and plums. The symptoms of plum pox disease or Sharka include fruit abnormality, loss of taste, and fruits that drop prematurely. These symptoms may cause losses as high as 80–100% of a crop and consequently it is a disease of economic significance in every country where stone fruits are grown. To achieve successful eradication or effective management of the associated disease, sensitive detection of the virus in infected plants is required.

PCR

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