Rapid and visual detection of milk vetch dwarf virus using recombinase polymerase amplification combined with lateral flow strips
Rapid and visual detection of milk vetch dwarf virus using recombinase polymerase amplification combined with lateral flow strips
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DOI：
10.1186/s12985-020-01371-5
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发表时间：
2020-07-11
期刊：
影响因子：
4.8
通讯作者：
Song, Xuemei
中科院分区：
文献类型：
--
作者：
Cao, Yuhao;Yan, Dankan;Song, Xuemei
背景紫云英矮缩病毒(Milk Vetch Dwarf Virus，MDV)是引起豆科植物黄化、矮化和卷叶症状的一种重要的单链DNA病毒。在中国，该病毒造成了巨大的经济损失，最近被发现感染烟草。其宿主范围的扩大和迅速传播的能力促使人们迫切需要一种敏感、特异和快速的诊断方法，以帮助有效地控制疾病。方法建立针对MDV外壳蛋白(CP)基因的聚合酶链式反应-侧流纸条检测(PCR-LFS)和重组酶聚合酶扩增-LFS(RPA-LFS)检测方法。结果琼脂糖凝胶电泳法可检测到10(3)拷贝和10(1)拷贝的MDV。建立的PCR-LFS和RPA-LFS方法都可以在37℃下检测到每次反应10(1)个拷贝，这两种方法都可以在粗叶提取物中检测到MDV。结论建立的RPA-LFS方法是一种快速、灵敏、特异的MDV检测方法，操作简便，具有较大的现场应用潜力。
Background Milk vetch dwarf virus (MDV) is an important ssDNA virus which causes yellowing, stunting and leaf rolling symptoms on legumes. In China, the virus causes great economic losses and has recently been found to infect tobacco. The expansion of its host range and its ability to spread rapidly has given rise to the urgent need for a sensitive, specific and rapid diagnostic assay that can assist in effective disease control. Methods Assays based on the polymerase chain reaction combined with lateral flow strip detection (PCR-LFS) and recombinase polymerase amplification combined with LFS (RPA-LFS) were developed targeting the coat protein (CP) gene of MDV. Results The PCR and RPA assays could detect respectively 10(3)copies or 10(1)copies of MDV by agarose gel electrophoresis. The PCR-LFS and RPA-LFS assays developed could both detect as few as 10(1)copies per reaction at 37 degrees C. Both methods could detect MDV in crude leaf extracts. Conclusions The RPA-LFS assay developed is a rapid, sensitive and specific method for detecting MDV, which is convenient and has great potential for use in the field.