摘要
A reverse-transcription loop-mediated isothermal amplification （RT-LAMP） method was established for the detection of wheat streak mosaic virus （WSMV）. Ac-cording to the conservative regions of the genes that encode the coat protein of WSMV, 2 pairs of primers were designed. Final y, the 1st pair of primers was select-ed through the specificity test. The sensitivity test showed the sensitivity of RT-LAMP method was 10 times higher than that of RT-PCR. In addition, the amplifica-tion of target gene could be judged visual y from the presence of fluorescence （cal-cein） in the final reaction system. The RT-LAMP method, established in this study, was rapid, easy, specific and sensitive. Moreover, it did not require sophisticated equip-ment. The RT-LAMP was suitable for the rapid detection of WSMV.
采用逆转录环介导等温扩增技术(reverse-transcription loop-mediated isothermal amplification,RT-LAMP),建立了小麦线条花叶病毒检测方法。根据WSMV外壳蛋白基因序列的保守区设计了2组引物,通过特异性试验筛选确定了第1组引物;灵敏度试验显示该方法的灵敏度比RT-PCR高10倍。此外,通过在反应体系中加入钙黄绿素,可在反应结束后通过裸眼观察荧光的有无来判断是否有扩增。该研究所建立的WSMV RT-LAMP方法具有快速、操作简单、特异性强、灵敏度高的特点,不需要复杂的仪器设备,适合于实验室现场快速检测。
