title: One-step reverse transcription loop-mediated isothermal amplification for the detection of Maize chlorotic mottle virus in maize.
abstract: 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 (RT-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 60min under isothermal condition at 63°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.
authors.affiliation: State Key Laboratory of Agro-biotechnology and Ministry of Agriculture Key Laboratory for Plant Pathology, China Agricultural University, Beijing 100193, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Agro-biotechnology and Ministry of Agriculture Key Laboratory for Plant Pathology, China Agricultural University, Beijing 100193, China.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Jiao
authors.affiliation: State Key Laboratory of Agro-biotechnology and Ministry of Agriculture Key Laboratory for Plant Pathology, China Agricultural University, Beijing 100193, China.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Liu
authors.affiliation: Beijing Entry-exit Inspection and Quarantine Bureau, Beijing 100016, China.
authors.forename: Xingliang
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Agro-biotechnology and Ministry of Agriculture Key Laboratory for Plant Pathology, China Agricultural University, Beijing 100193, China.
authors.forename: Zihao
authors.initials: Z
authors.lastname: Xia
authors.affiliation: Beijing Entry-exit Inspection and Quarantine Bureau, Beijing 100016, China.
authors.forename: Congliang
authors.initials: C
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Agro-biotechnology and Ministry of Agriculture Key Laboratory for Plant Pathology, China Agricultural University, Beijing 100193, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Agro-biotechnology and Ministry of Agriculture Key Laboratory for Plant Pathology, China Agricultural University, Beijing 100193, China. Electronic address: fanzf@cau.edu.cn.
authors.forename: Zaifeng
authors.initials: Z
authors.lastname: Fan
mesh_terms: D002681:China; D017931:DNA Primers; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013696:Temperature; D019183:Tombusviridae / Q000235:genetics* / Q000302:isolation & purification; D003313:Zea mays / Q000821:virology*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
keywords: Detection; Loop-mediated isothermal amplification (LAMP); Maize; Maize chlorotic mottle virus (MCMV)