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Sugarcane brown rust (SCBR), caused by <i>Puccinia melanocephala</i>, is a destructive fungal disease that has extensively spread in the sugarcane-cultivating regions across the world. Early monitoring plays an important role in predicting the <i>P. melanocephala</i> epidemic and managing SCBR. However, accurately identifying SCBR based on symptoms and urediniospore morphology at the initial stage is a challenge. Further, it is tedious, time-consuming, labor-intensive, and requires expensive equipment to detect <i>P. melanocephala</i> using PCR-based methods. Loop-mediated isothermal amplification (LAMP) technology is renowned for its speed, simplicity, and low equipment requirements for specifically and sensitively identifying many pathogens. Therefore, in this study, a novel and highly sensitive LAMP assay was developed for the specific detection of <i>P. melanocephala</i> in sugarcane. Here, the internal transcribed spacer (ITS) sequence of <i>P. melanocephala</i> was selected as the target gene for LAMP primer design. Based on the color change of SYBR Green I and gel electrophoresis, specific LAMP primers were screened. Further, the optimal reaction conditions for the LAMP assay were determined at 63 °C for 60 min. The LAMP assay showed a high degree of specificity for the detection of <i>P. melanocephala</i> in sugarcane, with no cross-reactivity with other fungal pathogens. The established LAMP protocol was highly sensitive and can be used to detect as low as 1 pg/μL of <i>P. melanocephala</i> plasmid DNA, which is comparable to that of nested PCR and ~100 times more sensitive than conventional PCR. Finally, the detection rate of the LAMP method was higher than that of conventional and nested PCR in field samples.