Citrus bark cracking viroid (CBCVd), formerly known as pathogen in the genus Citrus and first detected in Slovenian hops in 2014, threatens hop production as it leads to important economic losses. Reduction in yield and quality and even death of the infected plants within a few years are typical observations due to CBCVd infections of hops. The viroid is easily transmitted and spreads rapidly. As it cannot be controlled by plant protection measures, avoiding its introduction into hop gardens and eradicating first centres of infection are of utmost importance. An indispensable prerequisite is a reliable detection method suitable for large-scale routine testing. In this study, the development of primers and probe for real-time RT-PCR for sensitive CBCVd detection is described. To exclude “false negative” results, a nad5 mRNA-based internal positive control was included. To our knowledge, this is the first time such a duplex real-time RT-PCR detection method for CBCVd at least in hops is described. In addition, first method validation data are presented.

柑橘树皮裂化类病毒（CBCVd），以前称为柑橘属病原体并于2014年在斯洛文尼亚啤酒花中首次发现，它威胁酒花产量，因为这会造成重大的经济损失。由于酒花的CBCVd感染，典型的观察结果是几年内受感染植物的产量和质量下降甚至死亡。类病毒很容易传播并迅速传播。由于不能通过植物保护措施对其进行控制，因此至关重要的是避免将其引入啤酒花花园并消除主要的感染中心。必不可少的先决条件是一种适用于大规模常规测试的可靠检测方法。在这项研究中，描述了用于实时RT-PCR的敏感CBCVd检测引物和探针的开发。为了排除“假阴性”结果，包括了基于nad5 mRNA的内部阳性对照。据我们所知，这是首次至少在啤酒花中描述这种用于CBCVd的双工实时RT-PCR检测方法。此外，还介绍了第一种方法验证数据。