Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR. Validation and practice parameters Autors/ores Nieves Capote Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Edson Bertolini Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Antonio Olmos Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Eduardo Vidal Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia M. Carmen Martínez Valencian Institute for Agricultural Research (IVIA) Mariano Cambra Valencian Institute for Agricultural Research (IVIA) Paraules clau: Plum pox virus (PPV), sharka disease, spot real-time RT-PCR, winter or dormand diagnostic methods, post-test probability Resum Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PPV targets were amplified from fresh sections of plant tissues printed or squashed onto the same supports, without extract preparation. Spot real-time RT-PCR was validated as a PPV diagnostic method in samples collected during the dormancy period and showed high sensitivity (93.6%), specificity (98.0%), and post-test probability (97.9%) towards sharka disease. In an analysis of 2919 Prunus samples by spot real-time RT-PCR and DASI-ELISA 90.8% of the results coincided, demonstrating high agreement (k = 0.77 ± 0.01) between the two techniques. These results validate the use of immobilized PPV targets and spot real-time RT-PCR as screening method for largescale analyses. [Int Microbiol 2009; 12(1):1-6]. Biografies de l'autor/a Nieves Capote, Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Edson Bertolini, Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Antonio Olmos, Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Eduardo Vidal, Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia Valencian Institute for Agricultural Research (IVIA), Center for Plant Protection and Biotechnology, Valencia M. Carmen Martínez, Valencian Institute for Agricultural Research (IVIA) Valencian Institute for Agricultural Research (IVIA) Mariano Cambra, Valencian Institute for Agricultural Research (IVIA) Valencian Institute for Agricultural Research (IVIA) Descàrregues PDF (English) Publicat 2010-09-20 Número Vol. 12 Núm. 1 (2009) Secció Articles Llicència Submission of a manuscript to International Microbiology implies: that the work described has not been published before, including publication in the World Wide Web (except in the form of an Abstract or as part of a published lecture, review, or thesis); that it is not under consideration for publication elsewhere; that all the coauthors have agreed to its publication. The corresponding author signs for and accepts responsability for releasing this material and will act on behalf of any and all coauthors regarding the editorial review and publication process.If an article is accepted for publication in International Microbiology, the authors (or other copyright holder) must transfer to the journal the right–not exclusive–to reproduce and distribute the article including reprints, translations, photographic reproductions, microform, electronic form (offline, online) or any other reproductions of similar nature. Nevertheless, all article in International Microbiology will be available on the Internet to any reader at no cost. The journal allows users to freely download, copy, print, distribute, search, and link to the full text of any article, provided the authorship and source of the published article is cited. The copyright owner's consent does not include copying for new works, or resale. In these cases, the specific written permission of International Microbiology must first be obtained.Authors are requested to create a link to the published article on the journal's website. The link must be accompanied by the following text: "The original publication is available on LINK at <http://www.im.microbios.org>. Please use the appropiate URL for the article in LINK. Articles disseminated via LINK are indexed, abstracted, and referenced by many abstracting and information services, bibliographic networks, subscription agencies, library networks, and consortia.
