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

第一作者： Zeng, Rong

作者： Zeng, Rong;Xu, Li-hui;Gao, Shi-gang;Dai, Fu-ming;Ni, Xiu-hong;Chen, Chun-lei;Chen, Jian-cai

作者机构：

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：

页码：

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.

分类号： S432.1

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