逆转录环介导等温扩增技术检测南芥菜花叶病毒

Detection of Arabis mosaic virus by the reverse transcription loop-mediated isothermal amplification method

根据南芥菜花叶病毒外壳蛋白基因序列设计并合成了2组RT-LAMP引物,通过筛选试验,最终确定Ar4组引物为最佳引物,建立了南芥菜花叶病毒的RT-LAMP检测方法。同时,通过实时浊度仪和钙黄绿素分别对检测结果进行了判断。特异性和灵敏度试验结果显示,RT-LAMP方法快速、特异且灵敏,其灵敏度与普通RT-PCR法一致。

Two sets of primers were designed and synthesized based on coat protein gene of Arabis mosaic virus （ArMV） for reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay. Based on primer screening, the set Ar4 was selected as the optimal primers. A simple and sensitive RT-LAMP method was estab- lished to detect ArMV. Amplification products were detected by the real time turbidimeter and using calcein sepa- rately. Specificity and sensitivity assay showed that the RT-LAMP detection was rapid, specific and sensitive, and it was the same sensitive as RT-PCR.