Reverse transcription recombinase polymerase amplification (RT-RPA), an isothermal nucleic acid amplification and detection method, was developed to detect peach latent mosaic viroid (PLMVd) in pollen and peach leaves. Results showed that this RT-RPA detection method can be performed at 42 °C and completed in approximately 5 min, and there was no cross-reactivity with other common peach viruses. A sensitivity assay showed that the RT-RPA assay was 1000-fold more sensitive than a regular RT-PCR. Moreover, the method was successfully applied to test field-collected samples. The newly developed RT-RPA assay is rapid, sensitive, and reliable method for detection of PLMVd in peach pollen and leaves and can be utilized as an effective technique in quarantine and viroid-free certification processes.
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