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- W2521566254 abstract "El objetivo de este trabajo fue desarrollar un metodo para el diagnostico especifico del PepMoV basado en laRT-PCR. Se disenaron cebadores utilizando como referencia la secuencia genomica del aislado de la Florida, para amplificar un fragmento del extremo 5' no traducible y amino terminal de la proteina P1 del genoma. Se utilizo el programa Oligo (version 7.4) para realizar el analisis in silico y establecer los parametros iniciales de la reaccion. Se evaluaron la temperatura optima de hibridacion de cebadores, la concentracion de cebadores, el limite de deteccion, la repetitividad intraensayo e interensayo y la especificidad analitica. La temperatura optima de hibridacion de los cebadores fue de 59°C; se selecciono como concentracion optima de los cebadores 0,32 µM. Los aspectos evaluados del ensayo permitieron establecer las condiciones optimas de la tecnica. La RT-PCR se comporto con alta especificidad para el diagnostico de PepMoV; este mostro un limite de deteccion de 94 pg.µl-1. La herramienta molecular obtenida permitira apoyar los programas de mejoramiento genetico y de manejo integrado del cultivo en el pais. Palabras clave: RT-PCR, potyvirus, PepMoV, pimiento. Development of a method for the specific diagnosis of PepMoV based on RT -PCR ABSTRACT The main objective was to develop a method for the specific diagnosis of PepMoV based on RT-PCR. Primers were designed using as reference the genomic sequence of the Florida isolate to amplify a fragment of the 5'untranslatable end and amino-terminus of P1 protein of the genome. The Oligo program (version 7.4) was used to carry out the analysis in silico and establish the initial parameters of the reaction. The optimal temperature of primer hybridization, primer concentration, intra- and inter-assay repeatability, detection limit, and the analytic specificity were evaluated. The optimal temperature of primer hybridization was 59°C, and 0,32 µM was selected as the optimal concentration of the primers. The aspects of the assay evaluated allowed establishing the optimal conditions of the technique. The RT-PCR method behaved with a high specificity for the diagnosis of PepMoV. It showed a detection limit of 94 pg.µl-1. The molecular tool obtained will allow supporting the breeding programs and the integrated pest management of this crop in the country. Key words: RT-PCR, potyviruses, PepMoV, sweet pepper." @default.
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- W2521566254 date "2016-08-02" @default.
- W2521566254 modified "2023-09-23" @default.
- W2521566254 title "Desarrollo de un método para el diagnóstico específico del PepMoV basado en la RT-PCR" @default.
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