Emerg Infect Dis. 2022 Sep;28(9):1765-1769. doi: 10.3201/eid2809.220917. Epub 2022 Jul 29.
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.
自 2022 年 5 月以来，北半球的非猴痘流行国家报告了越来越多的猴痘病例。我们改编了 2 种已发表的实时定量 PCR 方法，用于在广泛使用的自动化高通量 PCR 系统上进行双靶点猴痘病毒检测。我们通过数字 PCR 对猴痘病毒参考物质进行连续稀释来确定分析性能，定量检测结果表明，联合检测的最低检测限为 4.795（95%CI3.6-8.6）拷贝/ml。我们使用临床剩余拭子样本，将我们的检测方法与一种商业的手动正痘病毒研究用 PCR 试剂盒进行了临床性能比较。我们的检测方法对 11 份阳性临床样本（n=11）和 56 份阴性临床样本（n=56）的检测结果均为阳性和阴性，完全符合。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。我们提供的检测方法简化了现有的广泛建立的 SARS-CoV-2 诊断检测平台上的猴痘病毒高通量分子检测。
