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• W2521375501 abstract "El objetivo de este trabajo fue evaluar las tecnicas de deteccion del Potato virus Y (PVY):TAS-ELISA e Inmunocaptura-RT-PCR en tiempo real (IC-RT-qPCR), en tejidos foliares de papa ( Solanum tuberosum L.) cv. Diacol-Capiro procedentes de tres lotes del Norte de Antioquia, Colombia. Adicionalmente, se secuencio el genoma de este virus mediante secuenciacion masiva de nueva generacion (NGS). Se detecto el PVY con ambas tecnicas en los tres lotes, aunque estas difirieron significativamente en su sensibilidad; con IC-RT-qPCR se encontro el virus en el 88,8% de las muestras, mientras que con TAS-ELISA en el 33,3%. La naturaleza de los amplicones se confirmo por secuenciacion Sanger (identidad=98-100%) y comparacion con los valores de temperaturas de fusion (Tm) del control positivo (77,5°C±1°C). Los analisis bioinformaticos permitieron caracterizar el genoma completo de la cepa PVY_Yarumal, con una profundidad de 716x y un tamano de 9701nt. Presenta un marco de lectura abierto (ORF) que codifica para una poliproteina de 3061 aminoacidos, flanqueada por regiones no traducidas (UTR) 5' y 3' de 187 y 328nt, respectivamente. Los analisis filogeneticos, tanto de la poliproteina completa como de la secuencia de la capside, indican su identidad con genotipos de la raza necrosante PVYNTN; se identificaron, ademas, los motivos K400E419 y QELA asociados a dicho linaje. Estos resultados plantean la necesidad de incorporar en los programas de manejo de enfermedades virales, en la agroindustria de papa en Colombia, el empleo de tecnicas de deteccion altamente sensibles como las basadas en RT-PCR en tiempo real. Palabras clave: ELISA, NGS, Potyvirus, RT-qPCR, Solanaceae. Detection and molecular characterization of Potato virus Y (PVY) in potato ( Solanum tuberosum L.) crops in northern Antioquia, Colombia ABSTRACT The aim of this work was to evaluate the techniques TAS-ELISA and IC-RT-qPCR as detection tools for Potato virus Y (PVY) in foliar tissues of potato ( Solanum tuberosum L.). Leaf samples of the potato variety Diacol-Capiro from three different plots in northern Antioquia (Colombia) were used. Additionally, the complete genome sequence of this virus was obtained using Next-generation sequencing (NGS). PVY detection using both techniques differed significantly in sensitivity, as 88.8% of the samples tested positive for PVY using IC-RT-qPCR in contrast to only 33.3% with TAS-ELISA. Sanger sequencing of the RT-qPCR amplicons and the comparison with the melting temperatures of the positive control (77.5°C±1°C) were used to confirm the presence of PVY. Bioinformatics analysis resulted in a contig of 9701 nt with an average sequence depth of 716x. The PVY_Yarumal contains an ORF coding for a 3061 residue protein flanked by 5' and 3' untranslated regions of 187 and 328 nt, respectively. Phylogenetic analysis using the complete polyprotein CDS and CP regions suggests phylogenetic identity with necrotic strain PVYNTN; sequence motifs K400E419and QELA typical of PVYNTN were also found in PVY_Yarumal. These results suggest that urgent measures are required to strengthen current integrated virus disease management in the potato agroindustry in Colombia by using highly sensitive detection techniques based on real-time RT-PCR. Key words: ELISA, NGS, Potyvirus, RT-qPCR, Solanaceae." @default.
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• W2521375501 date "2016-08-02" @default.
• W2521375501 modified "2023-09-23" @default.
• W2521375501 title "Detección y caracterización molecular del Potato virus Y (PVY) en cultivos de papa (Solanum tuberosum L.) del norte de Antioquia, Colombia" @default.
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