Department of Virology III, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan; Management Department of Biosafety, Laboratory Animals, and Pathogen Bank, National Institute of Infectious Disease, Musashimurayama, Tokyo, Japan.
Department of Virology III, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan.
J Virol Methods. 2023 Dec;322:114812. doi: 10.1016/j.jviromet.2023.114812. Epub 2023 Sep 21.
Human metapneumovirus (hMPV) is a common cause of respiratory infections in children. Many genetic diagnostic assays have been developed, but most detect hMPV regardless of the subgroup. In this study, we developed a real-time RT-PCR assay that can detect and identify the two major subgroups of hMPV (A and B) in one tube. Primers and probes were designed based on the sequences of recent clinical isolates in Japan. The assay showed comparable analytical sensitivity to a previously reported real-time RT-PCR assay and specific reactions to hMPV subgroups. The assay also showed no cross-reactivity to clinical isolates of 19 species of other respiratory viruses. In a validation assay using post-diagnosed clinical specimens, 98% (167/170) positivity was confirmed for the duplex assay, and the three specimens not detected were of low copy number. The duplex assay also successfully distinguished the two major subgroups for all 12 clinical specimens, for which the subgroup had already been determined by genomic sequencing analysis. The duplex assay described here will contribute to the rapid and accurate identification and surveillance of hMPV infections.
人偏肺病毒（hMPV）是儿童呼吸道感染的常见病因。已经开发出许多基因诊断检测方法，但大多数方法都可以检测到 hMPV，而不管亚群如何。在这项研究中，我们开发了一种实时 RT-PCR 检测方法，可以在一个管中同时检测和鉴定 hMPV 的两个主要亚群（A 和 B）。引物和探针是基于日本最近临床分离株的序列设计的。该检测方法与之前报道的实时 RT-PCR 检测方法具有相当的分析灵敏度，并且对 hMPV 亚群具有特异性反应。该检测方法也与其他 19 种呼吸道病毒的临床分离株无交叉反应。在使用确诊后的临床标本进行的验证检测中，双检测法的阳性率为 98%（167/170），未检测到的三个标本的拷贝数较低。该双检测法还成功区分了 12 个临床标本中的两个主要亚群，这些标本的亚群已经通过基因组测序分析确定。本文描述的双检测法将有助于快速准确地鉴定和监测 hMPV 感染。
