ORIGINAL ARTICLE
Multiplex RT-PCR reaction for simultaneous detection of Tomato torrado virus and Pepino mosaic virus co-infecting Solanum lycopersicum
 
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Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection – National Research Institute Władysława Węgorka 20, 60-318 Poznań, Poland
CORRESPONDING AUTHOR
Przemysław Wieczorek
Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection – National Research Institute Władysława Węgorka 20, 60-318 Poznań, Poland
Submission date: 2013-02-27
Acceptance date: 2013-07-30
 
Journal of Plant Protection Research 2013;53(3):289–294
KEYWORDS
TOPICS
ABSTRACT
The tomato (Solanum lycopersicum L.) is cultivated all over the world and is a vegetable of significant economic importance. However, an increased production of the vegetable is directly connected with an elevated occurrence of pathogens limiting the production efficiency of the vegetable. Both, Tomato torrado virus and Pepino mosaic virus have been found to be serious disease factors. When not controlled, these viruses can significantly decrease tomato cultivation. In this article, we report a multiplex reverse transcription-polymerase chain reaction (RT-PCR) protocol for simultaneous detection of both, Tomato torrado virus (ToTV) and Pepino mosaic virus (PepMV) in virus infected plants. The assay was designed to specifically amplify the conserved regions of genomic ribonucleic acid (RNA) of both viruses. Moreover, the glycerandehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control of amplification to exclude false-negative assay results. High-resolution melt analysis of generated RT-PCR products was additionally performed to increase sensitivity and double-check the specificity of the reaction without the need of subsequent complementary deoxyribonucleic acid (cDNA) sequencing.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
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