Detection of turnip mosaic virus (TuMV) in brassicaceous plants through colorimetric one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay
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Abstract
This study developed a highly efficient colorimetric one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for enhanced turnip mosaic virus (TuMV) detection. LAMP primers were newly designed targeting the coat protein (CP) gene of TuMV, ensuring specific amplification. The optimized protocol achieved successful amplification after a 45-min incubation in a heat block incubator at 65°C, with results immediately visible through distinctive colorimetric changes: positive samples transitioning from pink to yellowish color, whereas negative samples retained pink. Importantly, the sensitivity of this colorimetric one-step RT-LAMP significantly surpassed conventional RT-polymerase chain reaction (RT-PCR) technique for TuMV detection. The assay demonstrated complete specificity without cross-reactivity observed when tested against non-target plant viruses, confirming the high selectivity of the designed primers for TuMV detection. By integrating reverse transcription and amplification into a single reaction, this streamlined approach eliminates separate processing steps, substantially reducing both detection time and associated costs. This advancement represents a valuable tool for efficient diagnosis of RNA plant viruses in resource-limited settings.
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