中国农业科学2012，Vol.45Issue(7)：1285-1292,8.DOI:10.3864/j.issn.0578-1752.2012.07.006
水稻黑条矮缩病毒RT-LAMP快速检测方法的建立
Development of a RT-LAMP Assay for Rapid Detection of Rice black-streaked dwarf virus
摘要
Abstract
[Objective] The objective of this study is to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Rice black-streaked dwarf virus (RBSDV) from host plants and insect vector. [Method] Four primers matching a total of 6 sequences of the S10 of RBSDV were synthesized for the RT-LAMP assay. The concentration of the primer and MgSC>4 were optimized, and the best temperature and reaction time for detecting the virus were found, respectively. Detection sensitivity comparisons were performed between the RT-LAMP and RT-PCR assay using a single extraction of total RNA from RBSDV-infected rice leaves which was serially diluted in ten-fold increments in DEPC-treated water. In order to determine the specificity of the RT-LAMP assay, total RNA of rice leaves infected with RBSDV or Southern rice black-streaked dwarf virus (SRBSDV) was applied separately to the reaction system. RBSDV-infected rice plants collected from rice field were detected. [Result] This method demonstrated a high degree of specificity for RBSDV, which can distinguish RBSDV from SRBSDV. The method was also proved to be extremely sensitive, which was as much as the RT-PCR for RBSDV detection. The detection of amplified products was easily monitored. [Conclusion] The RT-LAMP assay is suitable for rapid detection of RBSDV in host plant and vectors.关键词
水稻黑条矮缩病毒/RT-LAMP/检测Key words
Rice black-streaked dwarf virus/RT-LAMP/detection引用本文复制引用
周彤,杜琳琳,范永坚,周益军..水稻黑条矮缩病毒RT-LAMP快速检测方法的建立[J].中国农业科学,2012,45(7):1285-1292,8.基金项目
国家公益性行业(农业)科研专项(201003031)、国家自然科学基金(31000841)、国家转基因生物新品种培育重大专项(2009ZX08001-013B)、江苏省农业科技自主创新资金项目(cx(10)414) （农业）
