Probert William S, Espinosa Alex, Hacker Jill K
Viral and Rickettsial Disease Laboratory, Center for Laboratory Sciences, California Department of Public Health, Richmond, CA, United States.
Front Public Health. 2025 Aug 1;13:1626030. doi: 10.3389/fpubh.2025.1626030. eCollection 2025.
The increasing incidence and global spread of mpox have prompted the World Health Organization to twice declare a Public Health Emergency of International Concern. Sustained human-to-human transmission, largely through sexual contact, and waning population immunity to smallpox have accelerated monkeypox virus evolution and driven the emergence of variants that can adversely affect the performance of existing molecular diagnostic tests. To minimize the risk of PCR target drop out and better detect and monitor emerging monkeypox virus variants, we have developed and validated a multiplex real-time PCR (MpoxEG4-plex rPCR) targeting highly conserved and essential orthopoxvirus genes for the detection of four analytes: orthopoxviruses, monkeypox virus, clade I monkeypox virus, and clade II monkeypox virus. The assay limit of detection was ≤ 9 genome copies per reaction, and the clinical accuracy, sensitivity, and specificity were > 96% for each analyte. The new assay was implemented to help confirm the first case of clade I mpox in the United States. The MpoxEG4-plex rPCR offers an accurate, informative, and reliable molecular diagnostic test for identifying cases and tracking case contacts in support of public health efforts to prevent and control the spread of mpox.
猴痘发病率的上升及其在全球的传播促使世界卫生组织两次宣布其为国际关注的突发公共卫生事件。主要通过性接触实现的持续人际传播以及人群对天花免疫力的下降加速了猴痘病毒的进化，并推动了可能对现有分子诊断检测性能产生不利影响的变体的出现。为了将聚合酶链反应（PCR）靶点缺失的风险降至最低，并更好地检测和监测新出现的猴痘病毒变体，我们开发并验证了一种多重实时PCR（MpoxEG4-plex rPCR），该方法针对高度保守且至关重要的正痘病毒基因，用于检测四种分析物：正痘病毒、猴痘病毒、I分支猴痘病毒和II分支猴痘病毒。该检测方法的检测限为每个反应≤9个基因组拷贝，每种分析物的临床准确性、敏感性和特异性均>96%。该新检测方法已用于协助确认美国首例I分支猴痘病例。MpoxEG4-plex rPCR为识别病例和追踪病例接触者提供了一种准确、信息丰富且可靠的分子诊断检测方法，以支持预防和控制猴痘传播的公共卫生工作。
