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Open Access Methodology

Rapid detection of wheat yellow mosaic virus by reverse transcription loop-mediated isothermal amplification

Zong-Ying Zhang, Xiao-Jun Liu, Da-Wei Li, Jia-Lin Yu and Cheng-Gui Han*

Author Affiliations

State Key Laboratory for Agrobiotechnology and Department of Plant Pathology, China Agricultural University, Beijing 100193, China

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Virology Journal 2011, 8:550  doi:10.1186/1743-422X-8-550

Published: 20 December 2011

Abstract

For the detection of wheat yellow mosaic virus (WYMV), we established a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. Using Primer Explorer software, four sets of primers were designed and RT-LAMP assay reaction conditions were optimized. The RT-LAMP was performed at different times by four primer sets. Agarose gel analysis showed that WYMV could be detected after 30 min with the primer set III and after 45 min with the other three primer sets, both under the 80-min reaction time. RT-LAMP had the same results with the four primer sets, thus primer set III and 65°C for 80 min reaction were selected for virus detection. There was no significant different when avian myeloblastosis virus (AMV) and moloney murine leukemia virus (M-MLV) RT-LAMP with the four primer sets and M-MLV was chosen due to its relatively cheap price. The result on specificity showed that the assay could amplify WYMV specifically, and the sensitivity comparison showed that the RT-LAMP was 100 times more sensitive than conventional reverse-transcriptase-polymerase chain reaction (RT-PCR). Overall, RT-LAMP was found to be a simple, specific, sensitive, convenient and time-saving method for WYMV detection.

Keywords:
Wheat yellow mosaic virus; RT-LAMP; Virus detection