University Medical Center Hamburg-Eppendorf (UKE), Institute of Medical Microbiology, Virology and Hygiene, Hamburg, Germany.
J Med Microbiol. 2021 Feb;70(2). doi: 10.1099/jmm.0.001295.
Laboratories worldwide are facing high demand for molecular testing during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, which might be further aggravated by the upcoming influenza season in the northern hemisphere. Given that the symptoms of influenza are largely indistinguishable from those of coronavirus disease 2019 (COVID-19), both SARS-CoV-2 and the influenza viruses require concurrent testing by RT-PCR in patients presenting with symptoms of respiratory tract infection. We adapted and evaluated a laboratory-developed multiplex RT-PCR assay for simultaneous detection of SARS-CoV-2 (dual target), influenza A and influenza B (SC2/InflA/InflB-UCT) on a fully automated high-throughput system (cobas6800). Analytical performance was assessed by serial dilution of quantified reference material and cell culture stocks in transport medium, including pretreatment for chemical inactivation. For clinical evaluation, residual portions of 164 predetermined patient samples containing SARS-CoV-2 (=52), influenza A (=43) or influenza B (=19), as well as a set of negative samples, were subjected to the novel multiplex assay. The assay demonstrated comparable analytical performance to currently available commercial tests, with limits of detection of 94.9 cp ml for SARS-CoV-2, 14.6 cp ml for influenza A and 422.3 cp ml for influenza B. Clinical evaluation showed excellent agreement with the comparator assays (sensitivity of 98.1, 97.7 and 100 % for Sars-CoV-2 and influenza A and B, respectively). The SC2/InflA/InflB-UCT allows for efficient high-throughput testing for all three pathogens and thus provides streamlined diagnostics while conserving resources during the influenza season.
全球实验室在严重急性呼吸综合征冠状病毒 2 型 (SARS-CoV-2) 大流行期间面临着分子检测的高需求，而北半球即将到来的流感季节可能会进一步加剧这种需求。鉴于流感的症状在很大程度上与 2019 年冠状病毒病 (COVID-19) 的症状无法区分，因此在出现呼吸道感染症状的患者中，需要同时进行 SARS-CoV-2 和流感病毒的 RT-PCR 检测。我们对一种实验室开发的用于同时检测 SARS-CoV-2（双靶标）、甲型流感和乙型流感的多重 RT-PCR 检测方法（SC2/InflA/InflB-UCT）进行了改编和评估，该方法在全自动高通量系统（cobas6800）上运行。通过在运输介质中对定量参考物质和细胞培养物进行连续稀释来评估分析性能，包括化学失活的预处理。在临床评估中，对包含 SARS-CoV-2(=52)、甲型流感(=43)或乙型流感(=19)的 164 个预定患者样本的剩余部分以及一组阴性样本进行了新的多重检测。该检测方法与当前可用的商业检测方法具有相当的分析性能，对 SARS-CoV-2 的检测限为 94.9 cp ml，对甲型流感的检测限为 14.6 cp ml，对乙型流感的检测限为 422.3 cp ml。临床评估显示与比较检测方法具有极好的一致性（SARS-CoV-2 和流感 A 和 B 的敏感性分别为 98.1%、97.7%和 100%）。SC2/InflA/InflB-UCT 可高效进行高通量检测这三种病原体，因此在流感季节可提供简化的诊断，同时节约资源。
