Detection of tomato mosaic virus infecting tomato using realtime RT-PCR
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Abstract
Tomato mosaic virus (ToMV) is known as one of the most common and devastating tomato viruses worldwide. It causes mosaic disease, which significantly impacts the productivity and quality of tomatoes in Vietnam. Early and accurate detection of ToMV in tomatoes is essential for effective disease control. This study developed a detection and quantification procedure for ToMV based on realtime RT-PCR. In this study, a positive control carrying ToMV’s target gene segment was amplified to a size of 595 bp, then cloned into pJET1.2 vector and transformed into Escherichia coli JM109. A realtime RT-PCR procedure was established using designed primers to amplify a 182 bp gene segment of the RdRP ORF2 gene region. A calibration curve was created with the equation y = -3.777x + 41.973, resulting in a correlation coefficient (R²) of 0.9939, which was used to quantify the ToMV virus. Additionally, the procedure quantified test samples with viral loads ranging from 1.7 x 104 to 9.5 x 106 copies/μL.
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References
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