Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR)
Viruses are important constraints to the movement and propagation of plant germplasm, especially for vegetatively propagated crops such as banana and plantain. Their control relies primarily on the use of virus-free plant material, whose production and certification requires sensitive and reliable detection methods. An existing polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) assay was adapted to the detection of Banana mild mosaic virus (BanMMV) and Banana virus X, two Flexiviridae infecting Musa spp. PDO inosine-containing primers were found to be well suited to the detection of BanMMV, despite its high molecular diversity, but not to that of the highly conserved BVX, for which species-specific primers were designed. Sampling and sample processing steps were optimized in order to avoid nucleic acid purification prior to the reverse transcription step. A polyclonal anti-BanMMV antiserum was raised and successfully used for the immunocapture (IC) of BanMMV viral particles from leaf extracts, leading to the development of a PDO-IC-RT-nested PCR assay. Although the anti-BanMMV antiserum could to some extent recognize BVX viral particles, direct binding (DB) was shown to be a more efficient method for processing BVX-infected samples and a PDO-DB-RT-nested PCR assay was developed for the detection of BVX from leaf extracts. © 2007 Elsevier B.V. All rights reserved.
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Subjects: | H20 - Maladies des plantes, F30 - Génétique et amélioration des plantes, Musa, virus des végétaux, PCR, Musa (bananes), Musa (plantains), http://aims.fao.org/aos/agrovoc/c_4993, http://aims.fao.org/aos/agrovoc/c_5985, http://aims.fao.org/aos/agrovoc/c_34079, http://aims.fao.org/aos/agrovoc/c_29128, http://aims.fao.org/aos/agrovoc/c_29127, |
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dig-cirad-fr-5390742024-01-28T15:12:24Z http://agritrop.cirad.fr/539074/ http://agritrop.cirad.fr/539074/ Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR). Teycheney Pierre-Yves, Acina Isabelle Nina, Lockhart Benham E.L., Candresse Thierry. 2007. Journal of Virological Methods, 142 (1-2) : 41-49.https://doi.org/10.1016/j.jviromet.2007.01.004 <https://doi.org/10.1016/j.jviromet.2007.01.004> Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) Teycheney, Pierre-Yves Acina, Isabelle Nina Lockhart, Benham E.L. Candresse, Thierry eng 2007 Journal of Virological Methods H20 - Maladies des plantes F30 - Génétique et amélioration des plantes Musa virus des végétaux PCR Musa (bananes) Musa (plantains) http://aims.fao.org/aos/agrovoc/c_4993 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_29128 http://aims.fao.org/aos/agrovoc/c_29127 Viruses are important constraints to the movement and propagation of plant germplasm, especially for vegetatively propagated crops such as banana and plantain. Their control relies primarily on the use of virus-free plant material, whose production and certification requires sensitive and reliable detection methods. An existing polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) assay was adapted to the detection of Banana mild mosaic virus (BanMMV) and Banana virus X, two Flexiviridae infecting Musa spp. PDO inosine-containing primers were found to be well suited to the detection of BanMMV, despite its high molecular diversity, but not to that of the highly conserved BVX, for which species-specific primers were designed. Sampling and sample processing steps were optimized in order to avoid nucleic acid purification prior to the reverse transcription step. A polyclonal anti-BanMMV antiserum was raised and successfully used for the immunocapture (IC) of BanMMV viral particles from leaf extracts, leading to the development of a PDO-IC-RT-nested PCR assay. Although the anti-BanMMV antiserum could to some extent recognize BVX viral particles, direct binding (DB) was shown to be a more efficient method for processing BVX-infected samples and a PDO-DB-RT-nested PCR assay was developed for the detection of BVX from leaf extracts. © 2007 Elsevier B.V. All rights reserved. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/539074/1/document_539074.pdf application/pdf Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1016/j.jviromet.2007.01.004 10.1016/j.jviromet.2007.01.004 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jviromet.2007.01.004 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1016/j.jviromet.2007.01.004 |
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H20 - Maladies des plantes F30 - Génétique et amélioration des plantes Musa virus des végétaux PCR Musa (bananes) Musa (plantains) http://aims.fao.org/aos/agrovoc/c_4993 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_29128 http://aims.fao.org/aos/agrovoc/c_29127 H20 - Maladies des plantes F30 - Génétique et amélioration des plantes Musa virus des végétaux PCR Musa (bananes) Musa (plantains) http://aims.fao.org/aos/agrovoc/c_4993 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_29128 http://aims.fao.org/aos/agrovoc/c_29127 |
spellingShingle |
H20 - Maladies des plantes F30 - Génétique et amélioration des plantes Musa virus des végétaux PCR Musa (bananes) Musa (plantains) http://aims.fao.org/aos/agrovoc/c_4993 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_29128 http://aims.fao.org/aos/agrovoc/c_29127 H20 - Maladies des plantes F30 - Génétique et amélioration des plantes Musa virus des végétaux PCR Musa (bananes) Musa (plantains) http://aims.fao.org/aos/agrovoc/c_4993 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_29128 http://aims.fao.org/aos/agrovoc/c_29127 Teycheney, Pierre-Yves Acina, Isabelle Nina Lockhart, Benham E.L. Candresse, Thierry Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) |
description |
Viruses are important constraints to the movement and propagation of plant germplasm, especially for vegetatively propagated crops such as banana and plantain. Their control relies primarily on the use of virus-free plant material, whose production and certification requires sensitive and reliable detection methods. An existing polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) assay was adapted to the detection of Banana mild mosaic virus (BanMMV) and Banana virus X, two Flexiviridae infecting Musa spp. PDO inosine-containing primers were found to be well suited to the detection of BanMMV, despite its high molecular diversity, but not to that of the highly conserved BVX, for which species-specific primers were designed. Sampling and sample processing steps were optimized in order to avoid nucleic acid purification prior to the reverse transcription step. A polyclonal anti-BanMMV antiserum was raised and successfully used for the immunocapture (IC) of BanMMV viral particles from leaf extracts, leading to the development of a PDO-IC-RT-nested PCR assay. Although the anti-BanMMV antiserum could to some extent recognize BVX viral particles, direct binding (DB) was shown to be a more efficient method for processing BVX-infected samples and a PDO-DB-RT-nested PCR assay was developed for the detection of BVX from leaf extracts. © 2007 Elsevier B.V. All rights reserved. |
format |
article |
topic_facet |
H20 - Maladies des plantes F30 - Génétique et amélioration des plantes Musa virus des végétaux PCR Musa (bananes) Musa (plantains) http://aims.fao.org/aos/agrovoc/c_4993 http://aims.fao.org/aos/agrovoc/c_5985 http://aims.fao.org/aos/agrovoc/c_34079 http://aims.fao.org/aos/agrovoc/c_29128 http://aims.fao.org/aos/agrovoc/c_29127 |
author |
Teycheney, Pierre-Yves Acina, Isabelle Nina Lockhart, Benham E.L. Candresse, Thierry |
author_facet |
Teycheney, Pierre-Yves Acina, Isabelle Nina Lockhart, Benham E.L. Candresse, Thierry |
author_sort |
Teycheney, Pierre-Yves |
title |
Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) |
title_short |
Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) |
title_full |
Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) |
title_fullStr |
Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) |
title_full_unstemmed |
Detection of Banana mild mosaic virus and Banana virus X by polyvalent degenerate oligonucleotide RT-PCR (PDO-RT-PCR) |
title_sort |
detection of banana mild mosaic virus and banana virus x by polyvalent degenerate oligonucleotide rt-pcr (pdo-rt-pcr) |
url |
http://agritrop.cirad.fr/539074/ http://agritrop.cirad.fr/539074/1/document_539074.pdf |
work_keys_str_mv |
AT teycheneypierreyves detectionofbananamildmosaicvirusandbananavirusxbypolyvalentdegenerateoligonucleotidertpcrpdortpcr AT acinaisabellenina detectionofbananamildmosaicvirusandbananavirusxbypolyvalentdegenerateoligonucleotidertpcrpdortpcr AT lockhartbenhamel detectionofbananamildmosaicvirusandbananavirusxbypolyvalentdegenerateoligonucleotidertpcrpdortpcr AT candressethierry detectionofbananamildmosaicvirusandbananavirusxbypolyvalentdegenerateoligonucleotidertpcrpdortpcr |
_version_ |
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