Division of Medical Microbiology, Department of Pathology, Johns Hopkins School of Medicine, Baltimore, Maryland, USA.
QIAGEN GmbH, Hilden, Germany.
J Clin Microbiol. 2024 May 8;62(5):e0002824. doi: 10.1128/jcm.00028-24. Epub 2024 Apr 19.
The mpox outbreak, caused by monkeypox virus (MPXV), accelerated the development of molecular diagnostics. In this study, we detail the evaluation of the Research Use Only (RUO) NeuMoDx MPXV assay by multiple European and US sites. The assay was designed and developed by Qiagen for the NeuMoDx Molecular Systems. Primers and probes were tested for specificity and inclusivity . The analytical sensitivity of the assay was determined by testing dilutions of synthetic and genomic MPXV DNA. A total of 296 clinical samples were tested by three sites; the Johns Hopkins University (US), UZ Gent (Belgium, Europe), and Hospital Universitario San Cecilio (Spain, Europe). The analytical sensitivity of the assay was 50 copies/mL for both clades I and II. The assay showed 100% identity for 80 clade I and 99.98% identity for 5,162 clade II genomes. Clade II primers and probes showed 100% specificity; however, identity of at least one of the two sets of clade I primers and probes with variola, cowpox, camelpox, and vaccinia viruses was noticed. The clinical validation showed sensitivity of 99.21% [95% confidence interval (CI): 95.66-99.98%] and specificity of 96.64% (95% CI: 91.62-99.08%) for lesion swab samples. The NeuMoDx MPXV Test shows acceptable analytical and clinical performance. The assay improves the laboratory's workflow as it consolidates nucleic acid extraction, PCR, data analysis, and interpretation and can be interfaced. The Test Strip can differentiate clades I and II, which has important laboratory safety implications.
In this manuscript, we provide detailed analysis and clinical evaluation of the assay using a large cohort of clinical samples across three academic centers in Europe and the United States. Because the assay differentiates MPXV clades I and II, this manuscript is timely due to the current need to rule out the regulated clade I by diagnostic clinical laboratories. In December 2023, and due to first report of cases of sexually transmitted clade I infections in the Democratic Republic of the Congo, when generic assays that do not differentiate the clades are used, samples are considered regulated. The assay meets the need of full automation and has a marked positive impact on the laboratory workflow.
猴痘病毒（MPXV）引起的猴痘疫情加速了分子诊断的发展。在这项研究中，我们详细介绍了由 Qiagen 为 NeuMoDx 分子系统设计和开发的 RUO NeuMoDx MPXV 检测在多个欧洲和美国站点的评估。检测针对特异性和包容性进行了测试。通过测试合成和基因组 MPXV DNA 的稀释液来确定检测的分析灵敏度。共有 296 个临床样本由三个地点进行了测试；约翰霍普金斯大学（美国）、UZ Gent（比利时，欧洲）和 Hospital Universitario San Cecilio（西班牙，欧洲）。该检测对两个分支 I 和 II 的分析灵敏度均为 50 拷贝/ml。该检测对 80 个分支 I 基因组的同一性为 100%，对 5162 个分支 II 基因组的同一性为 99.98%。分支 II 引物和探针显示出 100%的特异性；然而，分支 I 的两套引物和探针与天花、牛痘、骆驼痘和牛痘病毒中的至少一种具有相同的序列。临床验证显示，病变拭子样本的敏感性为 99.21%（95%置信区间：95.66-99.98%），特异性为 96.64%（95%置信区间：91.62-99.08%）。NeuMoDx MPXV 检测具有可接受的分析和临床性能。该检测通过整合核酸提取、PCR、数据分析和解释，并且可以进行接口，改善了实验室的工作流程。该检测能够区分 I 型和 II 型分支，这对实验室安全具有重要意义。
在本文中，我们使用来自欧洲和美国三个学术中心的大量临床样本对该检测进行了详细的分析和临床评估。由于该检测可区分 MPXV 分支 I 和 II，因此由于目前需要由诊断临床实验室排除受监管的分支 I，因此本文具有及时性。2023 年 12 月，刚果民主共和国首次报告了性传播的 I 型分支感染病例，当使用不能区分分支的通用检测时，这些样本被视为受监管的。该检测符合全自动化的需求，并对实验室工作流程产生了显著的积极影响。
