【摘 要】
：
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and optimized for detecting tomato chlorosis virus (ToCV), one of the most important viruses that infect to
【机 构】
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College of Plant Protection, Shandong Agricultural University, Tai'an 271018, China
【出 处】
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中国植物病理学会2014年学术年会
论文部分内容阅读
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and optimized for detecting tomato chlorosis virus (ToCV), one of the most important viruses that infect tomato crops worldwide.A set of four specific primers was designed in the RNA-dependent RNA polymerase (RdRp) gene.The RT-LAMP procedure could be completed within 40min under isothermal conditions of 60℃ without a thermal cycler, and no cross-reactivity was seen with other tomato viral pathogens.Sensitivity analysis showed that RT-LAMP could detect viral dilutions up to 2.0 × 10-7 g,which is 100-times more sensitive than reverse transcription-polymerase chain reaction (RT-PCR).In addition, naked-eye observation after staining in-tube RT-LAMP products with SYBR Green I facilitated detection of the presence of ToCV by avoiding the requirement for ethidium staining following gel electrophoresis.These results demonstrate that ToCV RT-LAMP is a rapid, sensitive, and affordable diagnostic tool that is more suitable than RT-PCR for the detection and surveillance of ToCV in field samples.
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