Differential detection of Wheat yellow mosaic virus, Japanese soil-borne wheat mosaic virus and Chinese wheat mosaic virus by reverse transcription loop-mediated isothermal amplification reaction

被引：38

作者：

Fukuta, Shiro ^{[1
]}

Tamura, Masaru ^{[2
]}

Maejima, Hidekazu ^{[3
]}

Takahashi, Reiko ^{[1
]}

Kuwayama, Sachiko ^{[1
]}

Tsuji, Takako ^{[1
]}

Yoshida, Tomofumi ^{[4
]}

Itoh, Koji ^{[4
]}

Hashizume, Hajime ^{[4
]}

Nakajima, Yasunori ^{[4
]}

Uehara, Yasushi ^{[3
]}

Shirako, Yukio ^{[4
]}

机构：

[1] Aichi Agr Res Ctr, Agroenvironm Div, Nagakute, Aichi 4801193, Japan

[2] Univ Tokyo, Asian Nat Environm Sci Ctr, Bunkyo Ku, Tokyo 1138657, Japan

[3] Nagano Agr Res Ctr, Nagano 3820051, Japan

[4] Aichi Agr Res Ctr, Field Crop Div, Nagakute, Aichi 4801193, Japan

来源：

JOURNAL OF VIROLOGICAL METHODS | 2013年 / 189卷 / 02期

关键词：

Differential detection; Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Wheat yellow mosaic virus (WYMV); Japanese soil-borne wheat mosaic virus ([!text type='JS']JS[!/text]BWMV); Chinese wheat mosaic virus (CWMV); Diagnosis; VISUAL DETECTION; LAMP REACTION; SEQUENCE; DNA;

D O I：

10.1016/j.jviromet.2013.03.005

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A differential detection method for three wheat viruses: Wheat yellow mosaic virus (WYMV), Japanese soil-borne mosaic virus (JSBWMV) and Chinese wheat mosaic virus (CWMV) using reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction was developed. All three primer sets, which were designed from the genome sequences of WYMV, JSBWMV and CWMV respectively, worked most efficiently at 65 degrees C and could detect each virus RNA within 10 min by fluorescence monitoring using an isothermal DNA amplification and fluorescence detection device. Furthermore, these primer sets showed unique annealing curves. The peak denaturing temperatures of WYMV, JSBWMV and CWMV primer sets were 87.6 degrees C, 84.8 degrees C and 86.4 degrees C, respectively and were clearly distinguished by the isothermal DNA amplification and fluorescence detection device. The RT-LAMP assay including all three primer sets was found to be 100 times more sensitive than RT-PCR for WYMV and JSBWMV and as sensitive as RT-PCR for CWMV. The RT-LAMP method was validated for the simultaneous detection of these viruses in wheat and barley leaves. (C) 2013 Elsevier B.V. All rights reserved.

引用

页码：348 / 354

页数：7

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