Beginning in May 2022, a rising number of monkeypox cases were reported in non–monkeypox-endemic countries in the Northern Hemisphere. We adapted 2 published quantitative PCRs for use as a dual-target monkeypox virus test on widely used automated high-throughput PCR systems. We determined analytic performance by serial dilutions of monkeypox virus reference material, which we quantified by digital PCR. We found the lower limit of detection for the combined assays was 4.795 (95% CI 3.6–8.6) copies/mL. We compared clinical performance against a commercial manual orthopoxvirus research use only PCR kit by using clinical remnant swab samples. Our assay showed 100% positive (n = 11) and 100% negative (n = 56) agreement. Timely and scalable PCR tests are crucial for limiting further spread of monkeypox. The assay we provide streamlines high-throughput molecular testing for monkeypox virus on existing broadly established platforms used for SARS-CoV-2 diagnostic testing.

从 2022 年 5 月开始，北半球非猴痘流行国家报告的猴痘病例数量不断增加。我们改编了 2 个已发表的定量 PCR，用作在广泛使用的自动化高通量 PCR 系统上的双靶点猴痘病毒测试。我们通过连续稀释猴痘病毒参考材料来确定分析性能，我们通过数字 PCR 对其进行量化。我们发现组合检测的检测下限为 4.795 (95% CI 3.6–8.6) 拷贝/mL。我们通过使用临床残余拭子样本将临床性能与仅使用 PCR 试剂盒的商业手动正痘病毒研究进行了比较。我们的测定显示 100% 阳性 (n = 11) 和 100% 阴性 (n = 56) 一致。及时且可扩展的 PCR 检测对于限制猴痘的进一步传播至关重要。