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Evaluation of the Use of One-Step Quantitative RT-PCR for Detection of Potato Virus X
Fig. 1.　Specificity evaluation of potato virus X (PVX) detection based on the reverse transcription-polymerase chain reaction (RT-PCR) (upper) and real-time RT-PCR (lower) assays. Amplification of PVX, pepper mild mottle virus (PMMV), tobacco mosaic virus (TMV), tomato mosaic virus (ToMV), potato virus Y (PVY), pepper mosaic virus (PMV), chili vein mosaic virus (CVMV), and pepper vein mottle virus (PVMV)-infected plants by RNA extracts. Health tomato samples as negative control (H-T). In RT-PCR, a 112-bp amplicon was generate only from positive control (PC) and PVX. Lane NTC: No Template Control and Lane M: 100 bp ladder.
Fig. 1.　Specificity evaluation of potato virus X (PVX) detection based on the reverse transcription-polymerase chain reaction (RT-PCR) (upper) and real-time RT-PCR (lower) assays. Amplification of PVX, pepper mild mottle virus (PMMV), tobacco mosaic virus (TMV), tomato mosaic virus (ToMV), potato virus Y (PVY), pepper mosaic virus (PMV), chili vein mosaic virus (CVMV), and pepper vein mottle virus (PVMV)-infected plants by RNA extracts. Health tomato samples as negative control (H-T). In RT-PCR, a 112-bp amplicon was generate only from positive control (PC) and PVX. Lane NTC: No Template Control and Lane M: 100 bp ladder.

Author:Cheng-Ping Kuan*, Ya-Ting Liu, Ssu-Yu Lin, Mei-Ju Lin, Shu Chen, and Yin-Hewey Cheng

Abstract:

This study developed a real-time reverse transcription-polymerase chain reaction (RT-PCR) detection method for potato virus X (PVX) by designing a specific primer pair based on the coat protein (CP) gene. The assay demonstrated a sensitivity approximately 10–100 times higher than conventional RT-PCR for PVX detection. Furthermore, it exhibited high specificity, accurately detecting PVX without cross-reacting with other solanaceous viruses or uninfected healthy plants. The reliability of this method was further confirmed through field testing on PVX-infected potato samples. Due to its high sensitivity and specificity, the real-time RT-PCR assay developed in this study has significant potential for the early detection of PVX infections in potatoes and tomatoes.

Key words:Potato virus X, Real-time RT-PCR, Detection

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Journal of
Taiwan Agricultural Research
Vol. 74, No.2 (2025) ISSN: 2790-086X Vol. 74, No.2 (2025)
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