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Multiple viral infections are discovered in papaya cultivation.All three of the viruses included in this study caused significant economic losses in papaya farming worldwide.The development of a SYBR Green I based multiplex real-time PCR is described for simultaneous detection of Papaya ringspot virus (PRSV), Papaya leaf distortion mosaic virus (PLDMV) and Papaya mosaic virus (PapMV) in papaya.Members infected different virus were identified by their distinctive melting temperatures (Tms): 81.0 ± 0.8 ℃ for PRSV isolates, 84.7 ± 0.6 ℃ for PLDMV isolates, and 88.7 ± 0.4 ℃ for PapMV isolates.