Diseño de una metodología de RT-PCR múltiplex para detección de virus en pitahaya Hylocereus spp

Abstract

CONDITION FOR PUBLICATION OF PROJECT. Pitaya (Hylocereus spp.) is a tropical fruit that has gained worldwide recognition in recent years. In Ecuador, it’s a product of increasing economic value that has positioned the country as one of the main exporters in South America. However, its production is limited by viral infections that can affect fruit yield and quality, especially those caused by the genera Potexvirus and Badnavirus. This project aims to establish a multiplex endpoint RT-PCR methodology for the simultaneous detection of prevalent viruses in pitaya. For this purpose, primers were designed and validated to detect both viruses, as well as a housekeeping gene (ACT7), which served as an internal control. On the other hand, protocol parameters related to the PCR reaction and thermocycling conditions were optimized. The developed methodology led to the amplification of both viruses and the reference gene in pitahaya samples. This scientific tool enables phytosanitary monitoring of crops, stimulates the availability of virus-free plant material for new production projects and contributes to reducing operational costs and waste in molecular diagnostic laboratories. Key words: viral infections, phytopathology, primers, diagnostic