One-step reverse transcription loop mediated isothermal amplification assay for sensitive and rapid detection of Cucurbit chlorotic yellows virus
文献类型： 外文期刊
作者： Wang, Zhenyue 1 ; Gu, Qinsheng 3 ; Sun, Hu 2 ; Li, Honglian 1 ; Sun, Bingjian 1 ; Liang, Xiangzhi 1 ; Yuan, Yuan 1 ; Liu, 1 ;
作者机构： 1.Henan Agr Univ, Coll Plant Protect, Zhengzhou 450002, Peoples R China
2.Henan Acad Agr Sci, Inst Plant Protect, Zhengzhou 450002, Peoples R China
3.Chinese Acad Agr Sci, Zhengzhou Fruit Res Inst, Zhengzhou 450009, Peoples R China
关键词： CCYV;RT-LAMP;Detection
期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）
ISSN： 0166-0934
年卷期： 2014 年 195 卷
页码：
收录情况： SCI
摘要： A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of Cucurbit chlorotic yellows virus (CCYV). In this procedure, a set of four primers matching a total of six sequences in the coat protein gene region of CCYV was synthesized for the RT-LAMP assay using total RNA extracted from CCYV-infected melon leaf tissues, and the optimum reaction temperature and assay time were determined. The sensitivity assay showed that the virus was detectable in RT-LAMP reactions at dilutions of 1 x 10(-11), which was 10(5) times more sensitive than the RT-PCR assay. The RT-LAMP assay for CCYV and Sweet potato chlorotic stunt virus (SPCSV) exhibited high specificity for CCYV. This simple and sensitive method has potential for detection of CCYV in samples collected in the field. (C) 2013 Elsevier B.V. All rights reserved.
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