Bluetongue virus is a double-stranded RNA virus with 10 genome segments. VP2 is the primary target for neutralising antibodies and defines the serotype. Today, more than 27 serotypes are known, 24 are defined as "classical", and new serotypes are under investigation. Beside group-specific BTV-genome detection, additional serotype characterisation is important for disease control and epidemiological investigations. Therefore, a low-density RT-qPCR array representing a panel of group- and serotype-specific assays, was combined with an internal control system. For BTV serotype detection, both published and the newly developed in-house PCR systems were combined. The different primer-probe-mixes were placed in advance into a 96-well plate stored at -20 °C until use. At the time of analysis, the only template RNA was added to the prepared primer-probe-mixes and heat denatured at 95 °C for 3 min. After cooling, the master mix was added to each well and the PCR could run for around 90 min. The presented low-density TaqMan-RT-qPCR array enables fast and precise characterisation of the BTV serotype in clinical cases. Furthermore, mixed infections can be easily identified. In addition, the newly developed low-density RT-qPCR-array can easily be adapted to novel BTV strain variants or extended for relevant differential diagnosis.

中文翻译：

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BlueTYPE-一种低密度TaqMan-RT-qPCR阵列，用于鉴定所有24种经典Bluetongue病毒血清型。

蓝舌病毒是具有10个基因组区段的双链RNA病毒。VP2是中和抗体的主要靶标并定义血清型。如今，已知超过27种血清型，其中24种被定义为“经典”，并且正在研究新的血清型。除了特定组的BTV基因组检测外，其他血清型特征对于疾病控制和流行病学研究也很重要。因此，将代表一组特定于群体和血清型的检测方法的低密度RT-qPCR阵列与内部控制系统结合使用。对于BTV血清型检测，将已发布和新开发的内部PCR系统结合在一起。预先将不同的引物-探针混合物放入放置在-20°C的96孔板中直至使用。在分析时，将唯一的模板RNA添加到制备的引物-探针混合物中，并在95°C下热变性3分钟。冷却后，将预混合物添加到每个孔中，PCR可以进行约90分钟。提出的低密度TaqMan-RT-qPCR阵列可在临床病例中快速准确地表征BTV血清型。此外，混合感染很容易识别。此外，新开发的低密度RT-qPCR阵列可轻松适应新型BTV株变种或扩展用于相关的鉴别诊断。混合感染很容易识别。此外，新开发的低密度RT-qPCR阵列可轻松适应新型BTV株变种或扩展用于相关的鉴别诊断。混合感染很容易识别。此外，新开发的低密度RT-qPCR阵列可轻松适应新型BTV株变种或扩展用于相关的鉴别诊断。