医学
胰腺癌
环介导等温扩增
病毒学
癌症
逆转录环介导等温扩增
分子生物学
荧光染料
内科学
基因
实时聚合酶链反应
生物
逆转录酶
核糖核酸
遗传学
DNA
作者
Denis Revskij,J. Runst,C. Umstätter,L. Ehlers,S. Rohde,R. Jaster
标识
DOI:10.1016/j.pan.2022.06.186
摘要
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 60 min 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.
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