Development of a reverse transcription loop-mediated isothermal amplification assay for the rapid detection of Pepper mottle virus
文献类型： 外文期刊
作者： Luo, Xiangwen 1 ; Zhang, Deyong 1 ; Zheng, Limin 1 ; Peng, Jing 1 ; Li, Fan 2 ; Zhang, Songbai 1 ; Liu, Yong 1 ;
作者机构： 1.Hunan Acad Agr Sci, Hunan Plant Protect Inst, Key Lab Pest Management Hort Crops Hunan Prov, Changsha 410125, Hunan, Peoples R China
2.Yunnan Agr Univ, Key Lab Agr Biodivers Pest Management, China Educ Minist, Kunming 650201, Peoples R China
关键词： field disease diagnosis;PepMoV;Potyvirus;RT-LAMP assay;RT-PCR
期刊名称：CANADIAN JOURNAL OF PLANT PATHOLOGY （ 影响因子：2.442； 五年影响因子：1.993 ）
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收录情况： SCI
摘要： Pepper mottle virus (PepMoV) is a widespread threat to vegetable crop production in the USA and south-east Asia. We describe the development of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to detect PepMoV. The RT-LAMP assay was based on a set of four primers that match a specific region of the coat protein gene in the PepMoV genome. The detection limit of conventional RT-PCR detection was 1.47 x 10(-4) mu g mu L-1 of cDNA, whereas RT-LAMP was 10 times more sensitive. Using RT-LAMP, PepMoV detection was also highly specific, showing no cross-activity with four other potyviruses. Sixty-nine field samples collected from symptomatic pepper plants growing in five South China provinces were tested for the presence of PepMoV infection by performing an RT-LAMP assay as well as a conventional RT-PCR. Both methods detected PepMoV in 18 samples, demonstrating that the PepMoV-specific RT-LAMP assay could be a useful alternative tool for the diagnosis and epidemiological surveillance of PepMoV infections. The RT-LAMP assay also has the advantages that it can be performed in a low-tech environment and is quicker and cheaper to perform than conventional RT-PCR.
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