Background In 2022, the global dissemination of mpox virus (MPXV) outside endemic regions prompted the expansion of diagnostic testing worldwide. This study assesses the performance characteristics of 5 real-time polymerase chain reaction (PCR) assays in detecting MPXV during the 2022 outbreak. Methods Clinical specimens collected from patients across Ontario, Canada, were tested on the following assays: RealStar Orthopoxyvirus PCR and FlexStar Monkeypox virus PCR (Altona Diagnostics), Novaplex MPXV (Seegene), VIASURE Monkeypox virus Real Time PCR Reagents (CerTest Biotec), and a laboratory-developed test. Positive percent agreement (PPA), negative percent agreement (NPA), relative limit of detection (LOD), and precision were evaluated and MPXV lineages were determined using an amplicon-based whole-genome sequencing (WGS) assay. Results Swabs were collected from various anatomic sites (65 positive and 30 negative). All assays demonstrated 100% NPA (95% confidence interval, 88.4%/88.1%–100.0%), with PPA ranging from 92.2% (82.7%–97.4%) to 96.9% (89.3%–99.6%). LOD and precision were comparable across assays, with coefficient of variations <3%. WGS analysis identified 6 lineages, all belonging to subclade IIb. Conclusions The assays exhibited excellent PPA, NPA, LOD, and precision. Ongoing performance monitoring is essential to detect assay escape mutants and ensure universal detection of evolving MPXV strains.

中文翻译：

---

5 种 Mpox 病毒检测聚合酶链式反应检测方法的评价

背景 2022 年，mpox 病毒 (MPXV) 在流行地区以外的全球传播促使诊断检测在全球范围内扩大。本研究评估了 2022 年爆发期间检测 MPXV 的 5 种实​​时聚合酶链反应 (PCR) 检测的性能特征。方法 对从加拿大安大略省各地患者采集的临床标本进行以下检测：RealStar Orthopoxyvirus PCR 和 FlexStar 猴痘病毒 PCR (Altona Diagnostics)、Novaplex MPXV (Seegene)、VIASURE 猴痘病毒实时 PCR 试剂 (CerTest Biotec) 和实验室开发的测试。评估了阳性一致性百分比 (PPA)、阴性一致性百分比 (NPA)、相对检测限 (LOD) 和精确度，并使用基于扩增子的全基因组测序 (WGS) 测定法确定了 MPXV 谱系。结果 从不同解剖部位采集拭子（65 个阳性，30 个阴性）。所有检测均显示 100% NPA（95% 置信区间，88.4%/88.1%–100.0%），PPA 范围为 92.2%（82.7%–97.4%）至 96.9%（89.3%–99.6%）。 LOD和精确度在测定中具有可比性，变异系数＜3％。全基因组测序分析确定了 6 个谱系，全部属于亚进化枝 IIb。结论 该检测显示出优异的 PPA、NPA、LOD 和精密度。持续的性能监测对于检测检测逃逸突变体并确保普遍检测不断进化的 MPXV 毒株至关重要。