Plant Protect. Sci., 2002, 38(10):S24-S27 | DOI: 10.17221/10313-PPS

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

D. Vašková, J. Špak

¹ Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, 370 05 České Budějovice, Czech Republic

² Faculty of Biological Sciences, University of South Bohemia, 370 05 České Budějovice, Czech Republic Tel.: +420 387 7775 521, Fax:+420 387 5300 356, E-mail: vaskova@umbr.cas.cz

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

Published: June 30, 2002  Show citation