Institute of Virology, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany.
J Med Microbiol. 2010 Jun;59(Pt 6):713-717. doi: 10.1099/jmm.0.017244-0. Epub 2010 Mar 4.
Although infections with the novel pandemic 2009 influenza A (H1N1) virus (A/H1N1/2009) appeared to be relatively mild during the first summer of circulation ('off season'), there has been significant morbidity and hospitalization and several fatal cases. Thus, rapid detection of A/H1N1/2009 is crucial for efficient treatment and infection control measures. In contrast to seasonal influenza, where point-of-care (POC) rapid antigen tests and direct fluorescent antibody (DFA) staining ensure rapid detection, diagnosis of A/H1N1/2009 has so far been based on RT-PCR. This study retrospectively compared the performance of the Quidel QuickVue POC test, DFA staining and virus isolation with that of RT-PCR for A/H1N1/2009 detection in 526 respiratory specimens collected during the first wave of the outbreak from May to September 2009. A/H1N1/2009 was detected in 9.1% (48/526) of samples. One hundred and thirty-seven of the A/H1N1/2009 PCR-negative samples were additionally tested using a RealAccurate Respiratory RT-PCR panel, revealing other respiratory viruses (mainly entero/rhino- and adenoviruses) in 42.3% (58/137). All methods analysed detected A/H1N1/2009 with excellent specificity but different sensitivities (POC test: 18.2%; DFA staining: 38.7%; virus isolation: 45.7%). Therefore, the POC test was not suitable for diagnosis, detecting A/H1N1/2009 only if present in high concentrations (corresponding median Ct value=19.0; range=16.5-21.4). DFA staining was also able to detect A/H1N1/2009 in specimens with a lower virus concentration (median Ct value=24.0; range=16.5-29.8). Virus isolation, which was positive after a median time of 7.5 days, was too time-consuming. In summary, DFA staining is superior to POC testing and may be appropriate for patients expected to have a rather high level of virus replication. Nevertheless, in DFA-negative specimens, A/H1N1/2009 should be excluded by RT-PCR.
虽然新型大流行的 2009 年甲型流感（H1N1）病毒（A/H1N1/2009）在首次传播的夏季（“淡季”）期间似乎相对较轻，但仍有大量发病率和住院率，并有几例死亡病例。因此，快速检测 A/H1N1/2009 对于有效治疗和感染控制措施至关重要。与季节性流感不同，即时检测（POC）快速抗原检测和直接荧光抗体（DFA）染色可确保快速检测，而 A/H1N1/2009 的诊断迄今一直基于 RT-PCR。本研究回顾性比较了 Quidel QuickVue POC 检测、DFA 染色和病毒分离在 2009 年 5 月至 9 月大流行第一波期间收集的 526 份呼吸道标本中检测 A/H1N1/2009 的性能。在 9.1%（48/526）的样本中检测到 A/H1N1/2009。对 137 份 A/H1N1/2009 PCR 阴性样本使用 RealAccurate Respiratory RT-PCR 试剂盒进行了进一步检测，结果在 42.3%（58/137）的样本中检测到其他呼吸道病毒（主要是肠病毒/鼻病毒和腺病毒）。所有分析方法均具有出色的特异性，但敏感性不同（POC 检测：18.2%；DFA 染色：38.7%；病毒分离：45.7%）。因此，POC 检测不适合诊断，仅在病毒浓度较高时（对应中位 Ct 值=19.0；范围=16.5-21.4）才能检测到 A/H1N1/2009。DFA 染色也能够检测到病毒浓度较低的 A/H1N1/2009（中位 Ct 值=24.0；范围=16.5-29.8）。病毒分离在中位时间为 7.5 天后才呈阳性，耗时太长。总之，DFA 染色优于 POC 检测，可能适用于预期病毒复制水平较高的患者。然而，在 DFA 阴性标本中，应通过 RT-PCR 排除 A/H1N1/2009。
