Johns Hopkins University, Department of Medicine, Baltimore, MD 21205, USA.
J Virol Methods. 2011 Apr;173(1):60-6. doi: 10.1016/j.jviromet.2011.01.007. Epub 2011 Jan 21.
Diagnosis of the etiologic agent of respiratory viral infection relies traditionally on culture or antigen detection. This pilot evaluation compared performance characteristics of the RT-PCR and electrospray ionization mass spectrometry (RT-PCR/ESI-MS) platform to conventional virologic methods for identifying multiple clinically relevant respiratory viruses in nasopharyngeal aspirates. The RT-PCR/ESI-MS respiratory virus surveillance kit was designed to detect respiratory syncytial virus, influenza A and B, parainfluenza types 1-4, adenoviridae types A-F, coronaviridae, human bocavirus, and human metapneumovirus. Patients (N=192) attending an emergency department during the 2007-2008 respiratory season consented, and "excess" frozen archived nasopharyngeal aspirates were analysed; 46 were positive by conventional virology and 69 by RT-PCR/ESI-MS, among which there were six samples with multiple viral pathogens detected. The sensitivity and specificity of the assay were 89.1% and 80.3%, respectively. Additional viruses that were not identified by conventional virology assays were detected (4 human bocaviruses and 7 coronaviruses). Samples in which the RT-PCR/ESI-MS results disagreed with conventional virology were sent for analysis by a third method using a commercial RT-PCR-based assay, which can identify viruses not detectable by conventional virologic procedures. Time to first result of RT-PCR/ESI-MS was 8h. RT-PCR/ESI-MS demonstrated capacity to detect respiratory viruses identifiable and unidentifiable by conventional methods rapidly.
呼吸道病毒感染的病因诊断传统上依赖于培养或抗原检测。本研究评估了 RT-PCR 和电喷雾电离质谱(RT-PCR/ESI-MS)平台与传统病毒学方法在鉴定鼻咽抽吸物中多种临床相关呼吸道病毒方面的性能特征。RT-PCR/ESI-MS 呼吸道病毒监测试剂盒旨在检测呼吸道合胞病毒、流感 A 和 B、副流感病毒 1-4 型、腺病毒 A-F 型、冠状病毒科、人博卡病毒和人偏肺病毒。在 2007-2008 年呼吸道季节期间,参加急诊科的患者同意,并分析了“多余”的冷冻存档鼻咽抽吸物;46 份经传统病毒学检测阳性,69 份经 RT-PCR/ESI-MS 检测阳性,其中有 6 份样本检测到多种病毒病原体。该检测的灵敏度和特异性分别为 89.1%和 80.3%。还检测到传统病毒学检测未鉴定的其他病毒(4 株人博卡病毒和 7 株冠状病毒)。与传统病毒学结果不一致的样本被送往使用商业基于 RT-PCR 的检测方法进行分析,该方法可鉴定传统病毒学方法不可检测的病毒。RT-PCR/ESI-MS 的第一个结果时间为 8 小时。RT-PCR/ESI-MS 能够快速检测传统方法可鉴定和不可鉴定的呼吸道病毒。