Improved diagnostic tools for the certification of strawberry propagation material - the use of PCR and NASBA for detection of Strawberry vein banding virus (SVBV)

D. Vašková and J. Špak
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D. Vašková: *Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, 370 05 České Budějovice, Czech Republic
J. Špak: *Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, 370 05 České Budějovice, Czech Republic

Plant Protection Science, 2002, vol. 38, issue SI1-6thConfEFPP, S24-S27

Abstract: Control of SVBV relies completely on the use of virus-free planting material, that can be tested either by biological indexing or by molecular methods. A NASBA-based amplification was developed for the detection of SVBV. NASBA is a method based on the primer-dependent, specific amplification of RNA by concurrent activity of a special enzyme mix (AMV-reverse transcriptase, RNaseH, T7 RNA polymerase) at a single temperature (41°C). Specific and sensitive detection of the amplified sequence can be performed in the same tube using molecular beacons. Sensitivity of SVBV-NASBA was 102 molecules of in vitro RNA detected per reaction. Results of the NASBA-based detection of SVBV in indicator strawberry plants were well comparable to the results of PCR.

Keywords: Strawberry vein banding virus; SVBV; Caulimovirus; Fragaria spp.; nucleic acid sequence based amplification; NASBA; molecular beacon (search for similar items in EconPapers)
Date: 2002
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DOI: 10.17221/10313-PPS

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