State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
J Virol Methods. 2010 Oct;169(1):211-4. doi: 10.1016/j.jviromet.2010.06.020. Epub 2010 Jul 14.
A reverse transcription loop-mediated isothermal amplification of DNA (RT-LAMP) for detection of Barley yellow dwarf viruses (BYDVs) was developed. In this procedure, three sets of four primers matching a total of six sequences of the coat protein or read-through protein genes of BYDVs - one each for three species, namely BYDV-GAV, -GPV and -PAV were synthesized for developing a specific and sensitive RT-LAMP assay for total RNA extracts from field-infected wheat plants in such a way that a loop could be formed and elongated during DNA amplification. RT-LAMP assays for each of three species of BYDV/CYDVs in China exhibited high specificity and could detect viral sequences in total RNA extracts from infected oat tissues at dilutions of 1 x 10(-5). All field samples collected from different regions of China showed the same result using both RT-LAMP and RT-PCR. This relatively simple and sensitive technique showed excellent potential with field-collected samples and for use in grassroots agencies in developing countries with limited resources.
一种逆转录环介导等温扩增 DNA（RT-LAMP）的方法被开发出来，用于检测大麦黄矮病毒（BYDVs）。在这个过程中，根据 BYDVs 外壳蛋白或通读蛋白基因的六个序列，总共设计了四组 6 个引物，每组对应 BYDV 的三个种，即 BYDV-GAV、-GPV 和 -PAV，以开发针对田间感病小麦总 RNA 提取物的特异性和敏感的 RT-LAMP 检测方法，以便在 DNA 扩增过程中形成和延伸环。针对中国三种 BYDV/CYDVs 的 RT-LAMP 检测均表现出高度特异性，可在 1 x 10(-5)的稀释度下从感染的燕麦组织总 RNA 提取物中检测到病毒序列。使用 RT-LAMP 和 RT-PCR 对从中国不同地区采集的所有田间样本进行检测，结果均相同。与使用传统的 RT-PCR 技术相比，这种相对简单和敏感的技术在采集的田间样本中具有很好的应用潜力，也适用于资源有限的发展中国家的基层机构。
