Diagnostic kits
database

Molecular

Molecular test

Real-Time PCR

The qPCR set enables the simultaneous detection (multiplex) of potato virus Y (PVY; all strains) and potato leafroll virus (PLRV) with real-time PCR. Viral RNA, extracted from potato samples, is amplified in a one-step RT-PCR reaction. The amplification of the cDNA can be monitored in real time, because the specific probes are labeled with fluorophores (PVY: FAM and PLRV: Cy5). An internal control (IC: JOE) is included for the convenience of the operator. The control shows whether the reverse transcription and the amplification of the RNA worked as intended.
Suitable tissue sources of viral RNA are potato leaves and tubers (peel, stolon, sprouts). It is highly recommended to use the BIOREBA “Potato DNA/RNA Rapid Extraction (included in the qPCR kits) for the extraction of RNA. The extraction method with Potato DNA/RNA rapid extraction has been validated and accredited according to ISO/IEC 17025. The method has been approved for the certification of seed potato. The qPCR for the detection of PLRV, PVY and IC was developed and validated in two certification seasons (2014/15) in collaboration with Agroscope, the Swiss centre of excellence for research in the agriculture and food sectors.

Leaves; peel; stolon; sprouts;

2

Potato leafroll virus (PLRV); potato virus Y (PVY)

It is highly recommended to use the BIOREBA “Potato DNA/RNA Rapid Extraction” (included in the qPCR kits) for the extraction of RNA. The extraction method with “Potato DNA/RNA rapid extraction” has been validated and accredited according to ISO/IEC 17025. The method has been approved for the certification of seed potato.

The qPCR set is validated for the simultaneous detection (multiplex) of potato virus Y (PVY), potato leaf roll virus (PLRV) and internal control (IC) in real- time PCR. Suitable tissues are potato leaves and tuber samples (peel, stolon and sprouts). Samples of up to 25 dormant tubers can be pooled for RNA extraction and analyzed according to this qPCR protocol.

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- Proficiency Tests organized by "Bayerische Landesanstalt für Landwirtschaft" in 2018, 2020 and 2021

(1) P. Kogovsek, L. Gow, M. Pompe-Novak, K. Gruden, G.D. Foster, N. Boonham, and M. Ravnikar 2008. Journal of Virological Methods 149:1-11.
(2) B.O. Agindotan, P.J. Shiel, and P.H. Berger 2007. Journal of Virological Methods 142:1-2.

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839610 (qPCR PLRV/PVY kit 96/10); 839625 (qPCR PLRV/PVY kit 96/25); 839210 (qPCR PLRV/PVY kit 192/10); 839225 (qPCR PLRV/PVY kit 192/25)

96 or 192 reactions (pools of 10 or 25)

839610 (qPCR PLRV/PVY kit 96/10)
Includes:
qPCR PLRV/PVY set 96
Potato DNA/RNA rapid extraction set 96/10x for 96 extractions (Pool size: up to 10 tuber samples)

839625 (qPCR PLRV/PVY kit 96/25)
Includes:
qPCR PLRV/PVY set 96
Potato DNA/RNA rapid extraction set 96/25x for 96 extractions (Pool size: up to 25 tuber samples)

839210 (qPCR PLRV/PVY kit 192/10)
Includes:
qPCR PLRV/PVY set 192
Potato DNA/RNA rapid extraction set 192/10x for 192 extractions (Pool size: up to 10 tuber samples)

839225 (qPCR PLRV/PVY kit 192/25)
Includes:
qPCR PLRV/PVY set 192
Potato DNA/RNA rapid extraction set 192/25x for 192 extractions (Pool size: up to 25 tuber samples)

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