Department of Plant Pathology, Washington State University, Prosser, WA, 99350, United States.
Department of Plant Pathology, Washington State University, Prosser, WA, 99350, United States.
J Virol Methods. 2019 Apr;266:25-29. doi: 10.1016/j.jviromet.2018.12.008. Epub 2019 Jan 14.
Apple stem grooving virus (ASGV) and Apple green crinkle-associated virus (AGCaV) negatively impact production, maintenance, and distribution of apples and other Malus species world-wide. Due to the increasing diversity of isolates found by high-throughput sequencing, we have developed real-time RT-qPCR assays for these two viruses. Primers and probes were designed against alignments of representative extant sequences from around the world, and reaction conditions optimized for sensitivity and specificity. Assays were validated against a panel of virus isolates, and compared to extant endpoint RT-PCR and ELISA assays. The new real-time RT-qPCR assays showed greater detection sensitivity than extant assays and were able to detect their target viruses from different host tissues.
苹果茎沟病毒（ASGV）和苹果绿皱果相关病毒（AGCaV）对苹果和其他苹果属植物的生产、维护和分布造成负面影响。由于高通量测序发现的分离株多样性不断增加，我们开发了这两种病毒的实时 RT-qPCR 检测方法。引物和探针是针对来自世界各地的代表性现有序列的比对设计的，并对反应条件进行了优化以提高灵敏度和特异性。该检测方法通过对一组病毒分离株进行验证，并与现有终点 RT-PCR 和 ELISA 检测方法进行比较。新的实时 RT-qPCR 检测方法比现有检测方法具有更高的检测灵敏度，并且能够从不同的宿主组织中检测到其目标病毒。
