Division of Clinical Pathology, Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan; Graduate Institute of Medical Science, National Defense Medical Center, Taipei, Taiwan.
Division of Clinical Pathology, Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
Int J Infect Dis. 2023 Feb;127:56-62. doi: 10.1016/j.ijid.2022.11.027. Epub 2022 Nov 28.
We have established a novel 5-in-1 VOC assay to rapidly detect SARS-CoV-2 and immediately distinguish whether positive samples represent variants of concern (VOCs).
This assay could distinguish among five VOCs: Alpha, Beta, Gamma, Delta, and Omicron, in a single reaction tube. The five variants exhibit different single nucleotide polymorphisms (SNPs) in their viral genome, which can be used to distinguish them. We selected target SNPs in the spike gene, including N501Y, P681R, K417N, and deletion H69/V70 for the assay.
The limit of detection of each gene locus was 80 copies per polymerase chain reaction. We observed a high consistency among the results when comparing the performance of our 5-in-1 VOC assay, whole gene sequencing, and the Roche VirSNiP SARS-CoV-2 test in retrospectively analyzing 150 clinical SARS-CoV-2 variant positive samples. The 5-in-1 VOC assay offers an alternative and rapid high-throughput test for most diagnostic laboratories in a flexible sample-to-result platform.
The assay can also be applied in a commercial platform with the completion of the SARS-CoV-2 confirmation test and identification of its variant within 2.5 hours.
我们建立了一种新型的五合一 VOC 检测方法，可快速检测 SARS-CoV-2 并立即区分阳性样本是否代表关注变异株（VOC）。
该检测方法可在单个反应管中区分五种 VOC：Alpha、Beta、Gamma、Delta 和 Omicron。这五种变体在其病毒基因组中存在不同的单核苷酸多态性（SNP），可用于区分它们。我们选择了 Spike 基因中的目标 SNP，包括 N501Y、P681R、K417N 和 H69/V70 缺失，用于该检测。
每个基因座的检测限为每个聚合酶链反应 80 个拷贝。我们通过对 150 例临床 SARS-CoV-2 变异阳性样本进行回顾性分析，比较了我们的五合一 VOC 检测、全基因测序和罗氏 VirSNiP SARS-CoV-2 检测的性能，发现结果高度一致。五合一 VOC 检测为大多数诊断实验室提供了一种替代的、快速的高通量检测方法，具有灵活的样本到结果平台。
该检测方法也可应用于商业平台，可在 2.5 小时内完成 SARS-CoV-2 确认检测并鉴定其变体。
