Schmallenberg virus (SBV), discovered in Germany in 2011, causes congenital malformations in ruminants. Reverse-transcription real-time PCR (RT-rtPCR) assays based on various segments of SBV have been developed for molecular detection. We developed alternative RT-rtPCR assays for SBV detection to avoid earlier reported mutations and hypervariable regions of the S and M segments of the viral genome. For SYBR Green-based detection of the S segment, the R² value and efficiency of the developed assay were 0.99 and 99%, respectively. For probe-based S segment detection, 2 assays were developed; the first had an R² value of 0.99 and 102% efficiency, and the second had a R² value of 0.98 and 86% efficiency. The probe-based M segment assay had an R² value of 1.00 and 103% efficiency. Detection limits of the RT-rtPCR assays with new primer sets were 10² and 10¹ copies/µL for the S and M segments, respectively. Field samples from cattle and sheep were also used for primary validation of the developed assays. Our assays should be suitable for SBV detection in ruminants and for in vitro studies of various SBV strains.

2011 年在德国发现的施马伦伯格病毒 (SBV) 会导致反刍动物先天性畸形。已经开发了基于 SBV 各个片段的逆转录实时 PCR (RT-rtPCR) 检测，用于分子检测。我们开发了用于 SBV 检测的替代 RT-rtPCR 检测，以避免早期报告的病毒基因组 S 和 M 片段的突变和高变区。对于 S 段的基于 SYBR Green 的检测，所开发测定的R ²值和效率分别为 0.99 和 99%。对于基于探针的 S 段检测，开发了 2 种检测方法；第一个的R ²值为 0.99 和 102% 的效率，第二个的R ²0.98 的值和 86% 的效率。基于探针的 M 片段测定具有1.00的R ²值和 103% 的效率。使用新引物组的RT-rtPCR 测定的检测限分别为10 ²和10 ¹拷贝/μL，用于S和M区段。来自牛和羊的田间样品也用于对开发的检测进行初步验证。我们的检测应适用于反刍动物的 SBV 检测和各种 SBV 菌株的体外研究。