Reverse Transcription–Recombinase Polymerase Amplification Assay for the Detection of Sugarcane Streak Mosaic Virus in Sugarcane
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Sugarcane streak mosaic virus (SCSMV, genus Poacevirus) causes sugarcane mosaic disease and leads to considerable economic losses. However, current methods for SCSMV diagnosis are limited. In this study, a novel isothermal assay was established on the basis of reverse transcription–recombinase polymerase amplification (RT-RPA) and evaluated in terms of its ability to detect SCSMV. The established RT-RPA assay with high specificity and reliability was 100-fold more sensitive than the RT-polymerase chain reaction. The evaluation of the temperature limit revealed that the assay had a flexible reaction temperature (26–46 °C), thereby indicating that the reaction mixture can be incubated with simple heating equipment, ambient temperature, or even human body heat. Time limit assessment results revealed that SCSMV can be detected in 5 min at 38 °C and in 20 min at 26 °C, thereby suggesting that the assay was rapid for SCSMV detection. Overall, the RT-RPA assay was specific, sensitive, reliable, and rapid for SCSMV detection, with a flexible and easily reachable reaction temperature. Thus, it can be an ideal method for SCSMV detection in sugarcane.
KeywordsRecombinase polymerase amplification Sugarcane Sugarcane mosaic disease Sugarcane streak mosaic virus
Enzyme-linked immunosorbent assay
Polymerase chain reaction
Recombinase polymerase amplification
Sugarcane streak mosaic virus
Sugarcane mosaic disease
Triticum mosaic virus
This study is supported by the National Natural Science Foundation of China (31771865) and the Sugar Crop Research System (CARS-170301).
SZ-Z conceived and designed the experiments. XY-F performed the experiments, result analysis, and manuscript drafting. LB-S, WZ-W, JG-W, and ZY-C revised the manuscript. All authors have read and approved the final manuscript.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
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