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RNA oligoprobe capture RT-PCR, a sensitive method for the detection ofGrapevine fanleaf virus in Tunisian grapevines

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Abstract

A simple, sensitive and specific RNA capture method is described for the detection of Grapevine fanleaf virus (GFLV) in infected grapevines. This method consists of hybridizing GFLV-RNAs to oligoprobes immobilized on nylon membranes, followed by RT-PCR amplification of targeted viral sequences. The RNA oligoprobe capture RT-PCR method is 10-fold more sensitive than IC-RT-PCR. The efficiency of the RNA oligoprobe capture RT-PCR and the reuse of immobilized oligoprobe membranes without loss of efficiency could make this procedure suitable for the routine diagnosis of GFLV in grapevines.

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Abbreviations

GFLV:

grapevine fanleaf virus

IC-RT-PCR:

immunocapture-RT-PCR

RT-PCR:

reverse transcription PCR

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Authors and Affiliations

  1. Laboratoire de Génie Biologique, Institut National des Sciences Appliquées et de Technologie, Centre Urbain Nord, BP No676, 1080, Tunis, Tunisie

    Sami Fattouch, Sonia M'hirsi & Nejib Marzouki

  2. Laboratoire de Génétique et Biologie Moléculaire, Faculté des Sciences de Tunis, Campus Universitaire, 1002, Tunisie

    Hajer Acheche & Mohamed marrakchi

Authors
  1. Sami Fattouch
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  2. Sonia M'hirsi
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  3. Hajer Acheche
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  4. Mohamed marrakchi
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  5. Nejib Marzouki
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Correspondence to Sami Fattouch.

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Fattouch, S., M'hirsi, S., Acheche, H. et al. RNA oligoprobe capture RT-PCR, a sensitive method for the detection ofGrapevine fanleaf virus in Tunisian grapevines. Plant Mol Biol Rep 19, 235–244 (2001). https://doi.org/10.1007/BF02772895

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