Rapid detection of papaya leaf distortion mosaic virus by a reverse transcription loop-mediated isot

来源 :中国热带作物学会遗传育种专业委员会2013年度学术研讨会 | 被引量 : 0次 | 上传用户:lovepc
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Papaya leaf distortion mosaic virus (PLDMV) can infect transgenic papaya resistant to a related pathogen, Papaya ringspot virus (PRSV), posing a substantial threat to papaya production in China.Current detection methods, however, are unable to be used for rapid detection in the field.Here, a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of PLDMV, using a set of four RT-LAMP primers designed based on the conserved sequence of PLDMV CP.The RT-LAMP method detected specifically PLDMV and was highly sensitive, with a detection limit of 1.32×10-6μg of total RNA per reaction.Indeed, the reaction was 10 times more sensitive than one-step RT-PCR, while also requiring significantly less time and equipment.The effectiveness of RT-LAMP and one-step RT-PCR in detecting the virus were compared using 90 field samples of non-transgenic papaya and 90 field samples of commercialized PRSV-resistant transgenic papaya from Hainan Island.None of the non-transgenic papaya tested positive for PLDMV using either method.In contrast, 19 of the commercialized PRSV-resistant transgenic papaya samples tested positive by RT-LAMP assay, and 6 of those tested negative by RT-PCR.Therefore, the PLDMV-specific RT-LAMP is a simple, rapid, sensitive, and cost-effective tool in the field diagnosis and control of PLDMV.
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