One-step reverse transcription loop-mediated isothermal amplification assay for rapid detection of melon yellow spot virus

被引：9

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
]}

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

来源：

EUROPEAN JOURNAL OF PLANT PATHOLOGY | 2016年 / 145卷 / 01期

关键词：

MYSV; RT-LAMP; Optimization; Visualization; TOSPOVIRUS; WATERMELON; THAILAND; JAPAN;

D O I：

10.1007/s10658-015-0821-6

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

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.

引用

页码：119 / 124

页数：6

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