One-step reverse transcription loop-mediated isothermal amplification assay for rapid detection of melon yellow spot virus
文献类型： 外文期刊
作者： Zeng, Rong 1 ; Xu, Li-hui 1 ; Gao, Shi-gang 1 ; Ni, Xiu-hong 2 ; Chen, Chun-lei 2 ; Chen, Jian-cai 2 ; Dai, Fu-ming 1 ;
作者机构： 1.Shanghai Acad Agr Sci, Inst Plant Protect, Shanghai Key Lab Protect Hort Technol, Shanghai 201403, Peoples R China
2.Pudong Dist Agrotechnol Extens Ctr, Shanghai 201300, Peoples R China
关键词： MYSV;RT-LAMP;Optimization;Visualization
期刊名称：EUROPEAN JOURNAL OF PLANT PATHOLOGY （ 影响因子：1.907； 五年影响因子：2.022 ）
ISSN：
年卷期：
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收录情况： SCI
摘要： A one-step, accelerated reverse transcription loop-mediated isothermal amplification (RT-LAMP) procedure was developed for the detection of Melon yellow spot virus (MYSV). The four primers required for accelerated RT-LAMP were designed to anneal onto a conserved region in MYSV N gene. Reaction temperature, optimal Mg2+ concentration, and the ratio of outer/inner primers affected reaction efficiency. MYSV RT-LAMP optimal reaction parameters were obtained in this study. The RT-LAMP was specific and sensitive in detecting MYSV in 60 minutes, with a detection limit of 0.5 x 10(-4) mu g total RNA per reaction, and a ladder-like pattern observed after agarose gel electrophoresis. RT-LAMP was 100-fold more sensitive than the conventional one-step reverse transcription polymerase chain reaction (RT-PCR) using serial dilutions of total RNA. RT-LAMP was a specific, sensitive, and low cost diagnostic tool that should be of value in more accurate determination of the distribution and host range of MYSV.
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