One-step reverse transcription loop mediated isothermal amplification assay for sensitive and rapid detection of Cucurbit chlorotic yellows virus

被引：31

作者：

Wang, Zhenyue ^{[1
]}

Gu, Qinsheng ^{[3
]}

Sun, Hu ^{[2
]}

Li, Honglian ^{[1
]}

Sun, Bingjian ^{[1
]}

Liang, Xiangzhi ^{[1
]}

Yuan, Yuan ^{[1
]}

Liu, Ruili ^{[1
]}

Shi, Yan ^{[1
]}

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

来源：

JOURNAL OF VIROLOGICAL METHODS | 2014年 / 195卷

关键词：

CCYV; RT-LAMP; Detection; 1ST REPORT; CUCUMBER;

D O I：

10.1016/j.jviromet.2013.08.037

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

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.

引用

页码：63 / 66

页数：4

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