Institute of Virology, Medical School Hannover, Hannover, Germany.
Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA.
J Med Virol. 2021 Jul;93(7):4438-4445. doi: 10.1002/jmv.26749.
The hologic panther fusion (PF) platform provides fully automated CE marked diagnostics for respiratory viruses, including the recently discovered severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) by a transcription mediated amplification (TMA) assay, but not for the endemic human coronaviruses (hCoV). Therefore, a laboratory developed test (LDT) comprising a multiplexed reverse transcription polymerase chain reaction (RT-PCR) protocol that detects and differentiates the four hCoV NL63, 229E, HKU1, and OC43 was adapted on the PF. The novel CE marked Aptima SARS-CoV-2 TMA and the LDT for hCoV were validated with 321 diagnostic specimens from the upper and lower respiratory tract in comparison to two SARS-CoV-2 RT-PCRs (PF E-gene RT-PCR and genesig RT-PCR, 157 specimens) or the R-GENE hCoV/hParaFlu RT-PCR (164 specimens), respectively. For the endemic hCoV, results were 96.3% concordant with two specimens discordantly positive in the PF and four specimens discordantly positive in the R-GENE assay. All discordantly positive samples had Ct values between 33 and 39. The PF hCoV LDT identified 23 hCoV positive specimens as NL63, 15 as 229E, 15 as HKU1, and 25 as OC43. The Aptima SARS-CoV-2 TMA gave 99.4% concordant results compared to the consensus results with a single specimen discordantly positive. Moreover, 36 samples from proficiency testing panels were detected and typed correctly by both novel methods. In conclusion, the SARS-CoV-2 TMA and the LDT for hCoV enhanced the diagnostic spectrum of the PF for all coronaviruses circulating globally for a multitude of diagnostic materials from the upper and lower respiratory tract.
Hologic 的 Panther Fusion (PF) 平台提供了完全自动化的 CE 标记诊断，用于呼吸道病毒，包括最近发现的严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2)，采用转录介导扩增 (TMA) 检测方法，但不用于地方性人类冠状病毒 (hCoV)。因此，在 Panther Fusion 上采用了一种实验室开发的测试 (LDT)，该测试由多重逆转录聚合酶链反应 (RT-PCR) 方案组成，用于检测和区分四种 hCoV NL63、229E、HKU1 和 OC43。新的 CE 标记 Aptima SARS-CoV-2 TMA 和用于 hCoV 的 LDT 通过与两种 SARS-CoV-2 RT-PCR（PF E 基因 RT-PCR 和 genesig RT-PCR，157 个样本）或 R-GENE hCoV/hParaFlu RT-PCR（164 个样本）进行比较，对来自上呼吸道和下呼吸道的 321 个诊断样本进行了验证。对于地方性 hCoV，结果与 PF 和 R-GENE 检测中各有两个样本不一致的结果具有 96.3%的一致性，有四个样本不一致的结果。所有不一致的阳性样本的 Ct 值均在 33 至 39 之间。PF hCoV LDT 确定了 23 个 hCoV 阳性样本为 NL63、15 个样本为 229E、15 个样本为 HKU1、25 个样本为 OC43。Aptima SARS-CoV-2 TMA 与共识结果相比具有 99.4%的一致性，仅有一个样本不一致。此外，两种新方法均可正确检测和分型来自能力验证试剂盒的 36 个样本。总之，SARS-CoV-2 TMA 和 hCoV 的 LDT 增强了 Panther Fusion 对全球循环的所有冠状病毒的诊断范围，适用于来自上呼吸道和下呼吸道的多种诊断材料。
