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Rapid detection of the rice false smut fungus Ustilaginoidea virens by lateral flow strip‑based recombinase polymerase amplification assay
Jiacheng Xi, Sanlian Wan, Yue Li, Yuandi Xu, Jing Yang, Ting Zhang, Jiajia Chen, Zhengguang Zhang, Danyu Shen, Haifeng Zhang
2024, 23 (11): 3763-3773.   DOI: 10.1016/j.jia.2023.09.027
Abstract117)      PDF in ScienceDirect      

Rice false smut, caused by Ustilaginoidea virens, is a devastating disease that greatly reduces rice yield and quality.  However, controlling rice false smut disease is challenging due to the unique infection mode of Uvirens.  Therefore, there is a need for early diagnosis and monitoring techniques to prevent the spread of this disease.  Lateral flow strip-based recombinase polymerase amplification (LF-RPA) overcomes the limitations of current Uvirens detection technologies, which are time-consuming, require delicate equipment, and have a high false-positive rate.  In this study, we used a comparative genomics approach to identify Uv_3611, a specific gene of Uvirens, as the target for the LF-RPA assay.  The designed primers and probe efffectively detected the genomic DNA (gDNA) of Uvirens and demonstrated no cross-reactivity with related pathogens.  Under optimal conditions, the LF-RPA assay demonstrated a sensitivity of 10 pg of Uvirens gDNA.  Additionally, by incorporating a simplified PEG-NaOH method for plant DNA extraction, the LF-RPA assay enabled the detection of Uvirens in rice spikelets within 30 min, without the need for specialized equipment.  Furthermore, the LF-RPA assay successfully detected Uvirens in naturally infected rice and seed samples in the field.  Therefore, the LF-RPA assay is sensitive, efficient, and convenient, and could be developed as a kit for monitoring rice false smut disease in the field.

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