Maize lethal necrosis disease (MLND) is a serious disease of worldwide importance. It is caused by the co-infection of maize with maize chlorotic mottle virus (MCMV) and a potyvirus, such as sugarcane mosaic virus (SCMV), that acts synergistically to produce more severe symptoms and production losses. More recently, maize yellow mosaic virus (MaYMV) and maize-associated totivirus (MATV) were found to co-infect with MCMV and SCMV in maize plants. To facilitate the detection of these viruses in co-infected maize, a multiplex RT-PCR assay was developed in this study. The assay used five specific primer pairs and simultaneously amplified these four viruses as well as the elongation factor 1α (EF 1α) gene use as internal control in one tube. The concentration of the primers, annealing temperature, annealing time, extension time and amplification cycles were optimized for the multiplex RT-PCR. The detection limit of the assay was up to 100 pg of total cDNA template. This multiplex RT-PCR assay was shown to be a sensitive and effective tool for the screening of field samples for the presence of these viruses in co-infected maize.

玉米致死性坏死病（MLND）是一种具有世界重要性的严重疾病。它是由玉米与玉米褪绿斑驳病毒 (MCMV) 和一种马铃薯病毒如甘蔗花叶病毒 (SCMV) 共同感染引起的，它们协同作用产生更严重的症状和生产损失。最近，发现玉米黄花叶病毒 (MaYMV) 和玉米相关全病毒 (MATV) 在玉米植物中与 MCMV 和 SCMV 共同感染。为了便于在共同感染的玉米中检测这些病毒，本研究开发了一种多重 RT-PCR 检测方法。该测定使用五对特异性引物，同时扩增这四种病毒以及延伸因子 1α ( EF 1α) 基因在一个试管中用作内部对照。针对多重 RT-PCR 优化了引物浓度、退火温度、退火时间、延伸时间和扩增循环。该测定的检测限高达总 cDNA 模板的 100 pg。这种多重 RT-PCR 测定被证明是一种敏感和有效的工具，用于筛选田间样品中这些病毒在共感染玉米中的存在。