The Application of Reverse Transcription-loop-mediated Isothermal Amplification for the Rapid Detection of Maize Chlorotic Dwarf Virus  

逆转录环介导等温扩增技术在玉米褪绿矮缩病毒快速检测中的应用（英文）

作　　者：徐颖[1] 张峰 于莹 邱志君[1] 

机构地区：[1]北仑出入境检验检疫局,浙江宁波315012

出　　处：《Agricultural Science & Technology》2017年第12期2450-2453,共4页农业科学与技术（英文版）

摘　　要：Maize chlorotic dwarf virus （MCDV） is a quarantine pest as approved by Chinese government. A rapid, sensitive and specific MCDV detection method using reverse transcription-loop-mediated isothermal amplification （RT-LAMP） was estab- lished in this study. Based on the sequence of MCDV coat protein coding gene, specific primers were designed and similar sensitivities were observed between RT- LAMP and RT-PCR, except that RT-LAMP was quicker, and the reaction could be finished within 1 h. In addition, the presence or absence of the fluorescent display in daylight allows naked easy detection of the amplification of MCDV genomic RNA using calcein. The RT-LAMP assay was applied successfully to detect MCDV in maize seeds, and the result by the addition of calcein was consistent with the result detected by the real time turbidimeter.玉米褪绿矮缩病毒(Maize chlorotic dwarf virus,MCDV)是我国对外公布的检疫性有害生物,该研究采用逆转录环介导等温扩增技术(Reverse transcription-loop-mediated isothermal amplification,RT-LAMP),建立了一种快速、灵敏和特异的MCDV检测方法。根据MCDV外壳蛋白基因序列的保守区设计特异性引物,灵敏度试验显示该方法的灵敏度与RTPCR方法相当,而检测时间明显缩短,1 h即可完成反应。此外,反应体系中加入钙黄绿素,可通过裸眼观察荧光的有无来判断是否有扩增。笔者还对玉米种子中携带的玉米褪绿矮缩病毒病毒进行了RT-LAMP检测,结果表明,裸眼判定与仪器监测结果一致。

关 键 词：Maize chlorotic dwarf virus （MCDV） Reverse transcription loop-mediatedisothermal amplification （RT-LAMP） Rapid detection 

分 类 号：S41-30[农业科学—植物保护]