International Journal of Applied Science and Technology

ISSN 2221-0997 (Print), 2221-1004 (Online) 10.30845/ijast

Molecular Diagnostic Assay for the Simultaneous Detection of Pome Fruit Viruses
In-Sook Cho, Gug-Seoun Choi, Sun-Jung Kwon

Multiplex RT-PCR assay was developed for the simultaneous detection of internal controls and viruses infecting apple and pear trees. These viruses are the most commonly occurring and economically important pathogens in apple and pear commercial orchards in Korea: Apple stems grooving virus (ASGV), Apple stem pitting virus (APSV), Apple chlorotic leaf spot virus (ACLSV) and/or Apple mosaic virus (ApMV). Primer concentrations and PCR conditions were optimized for the effective application of the assay. The sensitivity of detection by multiplex RT-PCR assay was up to 100 or 10-1 dilution depending on the virus. The multiplex RT-PCR assay was able to successfully detect target viruses infecting apple and pear. In addition, positive control plasmid was developed as controls for multiplex RT-PCR assay. Positive control plasmid was constructed by cloning sequences of target viruses infecting apple tree into a single vector. The multiplex RT-PCR assay developed is rapid and cost-effective molecular diagnosis tools for simultaneous detection of pome fruit viruses. The positive control plasmid is a new approach to apply as alternatives to classically used control infected plants for the detection of pome fruit viruses.

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