Hopkins Mark J, Redmond Catherine, Shaw Janet M, Hart Ian J, Hart C Anthony, Smyth Rosalind L, Semple Malcolm G
Liverpool Specialist Virology Centre, Royal Liverpool University Hospital, Liverpool, UK.
J Clin Virol. 2008 Jul;42(3):273-9. doi: 10.1016/j.jcv.2008.03.011. Epub 2008 May 2.
Human metapneumovirus (hMPV) causes a spectrum of respiratory disease ranging from trivial coryzal symptoms to fatal pneumonia, with a predilection for the very young, the immune suppressed and the frail elderly. Five distinct lineages of the virus genome have been described.
To develop and evaluate a sensitive, real-time PCR (RT-PCR) assay capable of detecting all lineages of hMPV, suitable for use in a diagnostic laboratory.
An RT-PCR assay was developed using novel primers and dual-labelled minor-groove-binding (MGB) probes complementary to consensus sequences. The assay and two alternative assays were tested against external quality assurance (EQA) panels. 221 respiratory samples collected during 2003-2004 were screened using the new assay. hMPV positive samples were sequenced and phylogenetically analysed.
Three genetic lineages of hMPV were detected during 2003-2004. Incidence was low (2.3%) compared to previous years. All five lineages had been present in the same community within the past 3 years.
The new assay correctly identified more EQA samples, including those at greatest dilution, than the alternative assays and detected all five lineages. Seasonal circulation of hMPV in paediatric patients with acute respiratory symptoms is dynamic with respect to incidence and viral genotype.
人偏肺病毒(hMPV)可引发一系列呼吸道疾病,症状从轻微的鼻卡他症状到致命性肺炎不等,该病毒尤其容易感染婴幼儿、免疫功能低下者和体弱的老年人。已描述了该病毒基因组的五个不同谱系。
开发并评估一种灵敏的实时聚合酶链反应(RT-PCR)检测方法,该方法能够检测hMPV的所有谱系,适用于诊断实验室。
使用新型引物和与共有序列互补的双标记小沟结合(MGB)探针开发了一种RT-PCR检测方法。该检测方法和另外两种替代检测方法针对外部质量保证(EQA)样本进行了测试。使用新的检测方法对2003年至2004年期间收集的221份呼吸道样本进行了筛查。对hMPV阳性样本进行测序并进行系统发育分析。
在2003年至2004年期间检测到hMPV的三个基因谱系。与前几年相比,发病率较低(2.3%)。在过去3年中,所有五个谱系都曾出现在同一社区。
与替代检测方法相比,新的检测方法能够正确鉴定更多的EQA样本,包括稀释度最高的样本,并且检测到了所有五个谱系。患有急性呼吸道症状的儿科患者中hMPV的季节性流行在发病率和病毒基因型方面具有动态变化。