Testing for potato viruses is globally very important to prevent a critical shortage of potato supply. In most countries, testing is obligated by law. In Germany, seed potatoes are monitored for six viruses: PLRV, PVY, PVM, PVA, PVX and PVS. They can cause up to 90% loss of potato tubers in the field. Common methods currently used for testing are ELISA and conventional real-time PCR, but both are very time-consuming, and the former needs a high capacity of green houses and human resources, the latter elaborate RNA extraction steps. Recently, we proposed a new method called real-time DiRT-PCR which enables us to test for PLRV, PVY and PVS along with an internal control in three duplex real-time PCR reactions directly on diluted tuber sap. In this study, we describe the first TaqMan® assay for PVM published so far and embed it into a multiplex system to detect the remaining viruses. We are now able to sensitively test for the presence of six viruses in two multiplex reactions using the real-time DiRT-PCR without RNA purification.

马铃薯病毒检测在全球范围内对于防止马铃薯供应严重短缺非常重要。在大多数国家，测试是法律规定的。在德国，监测种薯中的六种病毒：PLRV、PVY、PVM、PVA、PVX 和 PVS。它们可导致田间马铃薯块茎损失高达 90%。目前常用的检测方法有ELISA和常规的real-time PCR，但两者都非常耗时，并且前者需要大量的温室和人力资源，后者需要复杂的RNA提取步骤。最近，我们提出了一种称为实时 DiRT-PCR 的新方法，它使我们能够直接在稀释的块茎汁液上测试 PLRV、PVY 和 PVS 以及三个双重实时 PCR 反应中的内部对照。在这项研究中，我们描述了迄今为止发布的第一个用于 PVM 的 TaqMan® 检测，并将其嵌入到多路系统中以检测剩余的病毒。我们现在能够使用实时 DiRT-PCR 在两个多重反应中灵敏地测试六种病毒的存在，而无需纯化 RNA。