Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, De Pintelaan 185, 2P8, 9000, Ghent, Belgium.
Arch Virol. 2010 Feb;155(2):207-15. doi: 10.1007/s00705-009-0573-8. Epub 2009 Dec 15.
Human metapneumovirus (hMPV) is responsible for outbreaks of bronchiolitis in winter and early spring in young children. Due to the relatively recent discovery of hMPV, the diagnostic opportunities are limited, while differential diagnosis with respiratory syncytial virus (RSV) remains important. We validated the RT-PCR by comparing various methods of RNA extraction, one-step RT-PCR kits and primer-probe combinations. The optimized RT-PCR was evaluated using 47 nasopharyngeal aspirates (NPAs) collected from children younger than 5 years, with clinically suspected RSV infection. The evaluated RT-PCRs were also compared to a commercially available hMPV enzyme immunoassay (EIA). We found 8.5% hMPV positivity with both RT-PCRs, in agreement with published literature. hMPV EIA showed positive and indeterminate results in 17% and 8.5%, respectively, of the tested NPAs. Positive RT-PCR samples were positive or indeterminate by hMPV EIA. Samples that were positive for RSV and influenza A virus interfered with the hMPV EIA. In conclusion, although RT-PCR is already a valuable tool for diagnosing hMPV infections, further optimization of the RT-PCR method is recommended. The hMPV EIA kit shows poor specificity and therefore needs further improvement.
人偏肺病毒（hMPV）是导致婴幼儿冬春季节毛细支气管炎流行的病原体。由于 hMPV 是相对较新发现的病原体，因此诊断机会有限，而与呼吸道合胞病毒（RSV）的鉴别诊断仍然很重要。我们通过比较不同的 RNA 提取方法、一步法 RT-PCR 试剂盒和引物探针组合，对 RT-PCR 进行了验证。使用从 5 岁以下有临床疑似 RSV 感染的儿童中收集的 47 份鼻咽抽吸物（NPA）评估了优化的 RT-PCR。评估的 RT-PCR 也与市售的 hMPV 酶免疫测定（EIA）进行了比较。我们发现，两种 RT-PCR 的 hMPV 阳性率均为 8.5%，与已发表的文献一致。hMPV EIA 在 17%和 8.5%的测试 NPA 中分别显示阳性和不确定结果。阳性 RT-PCR 样本的 hMPV EIA 结果为阳性或不确定。RSV 和流感 A 病毒阳性样本会干扰 hMPV EIA。总之，尽管 RT-PCR 已经是诊断 hMPV 感染的有价值的工具，但建议进一步优化 RT-PCR 方法。hMPV EIA 试剂盒特异性差，因此需要进一步改进。
Zotero 插件