期刊
JOURNAL OF VIROLOGICAL METHODS
卷 240, 期 -, 页码 49-53出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2016.11.012
关键词
Maize chloro tic mottle virus (MCMV); Loop-mediated isothermal amplification (LAMP); Maize; Detection
资金
- Ministry of Agriculture [2014ZX08003-001]
- Ministry of Education (the 111 Project) [B13006]
- Special Fund for Detecting Plant Viruses from the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China [201310068, 2012BAK11B02]
Maize chlorotic mottle virus (MCMV) is spreading in many regions worldwide, causing maize lethal necrosis when co-infected with a potyvirid. In this study, one-step reverse transcription loop-mediated isothermal amplification (RI-LAMP) assay was developed to detect MCMV in maize. A set of four specific primers was designed based on the conserved coat protein gene sequences of MCMV. The RT-LAMP could be completed within 60 min under isothermal condition at 63 degrees C. The sensitivity test showed that the RT-LAMP was about 10-fold more sensitive than RT-PCR and no cross-reactivity was detected with other viral pathogens infecting maize in China. Moreover, the results of RT-LAMP could be visually inspected by SYBR Green I staining in a closed-tube, facilitating high-throughput application of MCMV detection. This method was further verified by testing field-collected samples. These results suggested that the developed MCMV RT-LAMP technique is a rapid, efficient and sensitive method which could be used as a routine screen for MCMV infection. (C) 2016 Elsevier B.V. All rights reserved.
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