RespiFinder: a new multiparameter test to differentially identify fifteen respiratory viruses
RespiFinder: a new multiparameter test to differentially identify fifteen respiratory viruses
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DOI：
10.1128/jcm.02294-07
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发表时间：
2008-04-01
影响因子：
9.4
通讯作者：
Simons, Guus
中科院分区：
文献类型：
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作者：
Reijans, Martin;Dingemans, Gijs;Simons, Guus
由于潜在传染源的数量仍在增加，对呼吸道感染患者的病原体进行广谱分析变得越来越重要。在这里，我们描述了新的多参数呼吸测定，这是基于多重连接依赖的探针扩增（MLPA）技术。该检测试剂盒可在一次反应中检测15种呼吸道病毒。MLPA反应之前是预扩增步骤，其确保以与单个单重实时逆转录PCR相同的特异性和灵敏度检测RNA和DNA病毒。使用144份临床样本对呼吸道合胞病毒检测试剂盒进行了验证，并将检测结果与细胞培养和呼吸道合胞病毒（RSV）特异性免疫层析检测试剂盒（伊卡）的结果进行了比较。与细胞培养结果相比，Respiratory试验对腺病毒的特异性和灵敏度分别为98.2%和100%;对人偏肺病毒的特异性和灵敏度分别为96.4%和100%;对甲型流感病毒（InfA）的特异性和灵敏度分别为98.2%和100%;对I型副流感病毒（PIV-1）的特异性和灵敏度分别为99.1%和100%; PIV-3分别为99.1%和80%;鼻病毒分别为90.1%和100%; RSV分别为94.6%和100%。与RSV特异性伊卡的结果相比，呼吸道感染试验的特异性和敏感性分别为82.4%和80%。在检测的临床标本中未检出PIV-2、PIV-4、B型流感病毒、InfA H5 N1和冠状病毒229 E。与通过细胞培养获得的诊断产率相比，使用呼吸道试验导致诊断产率增加（诊断产率分别为60%和35.5%）。总之，Respiratory assay提供了一种用户友好的高通量工具，可同时检测15种呼吸道病毒，具有出色的总体性能统计数据。
Broad-spectrum analysis for pathogens in patients with respiratory tract infections is becoming more relevant as the number of potential infectious agents is still increasing. Here we describe the new multiparameter RespiFinder assay, which is based on the multiplex ligation-dependent probe amplification (MLPA) technology. This assay detects 15 respiratory viruses in one reaction. The MLPA reaction is preceded by a preamplification step which ensures the detection of both RNA and DNA viruses with the same specificity and sensitivity as individual monoplex real-time reverse transcription-PCRs. The RespiFinder assay was validated with 144 clinical samples, and the results of the assay were compared to those of cell culture and a respiratory syncytial virus (RSV)-specific immunochromatography assay (ICA). Compared to the cell culture results, the RespiFinder assay showed specificities and sensitivities of 98.2% and 100%, respectively, for adenovirus; 96.4% and 100%, respectively, for human metapneumovirus; 98.2% and 100%, respectively, for influenza A virus (InfA); 99.1% and 100%, respectively, for parainfluenza virus type I (PIV-1); 99.1% and 80%, respectively, for PIV-3; 90.1% and 100%, respectively, for rhinovirus; and 94.6%-and 100%, respectively, for RSV. Compared to the results of the RSV-specific ICA, the RespiFinder assay gave a specificity and a sensitivity of 82.4% and 80%, respectively. PIV-2, PIV-4, influenza B virus, InfA H5N1, and coronavirus 229E were not detected in the clinical specimens tested. The use of the RespiFinder assay resulted in an increase in the diagnostic yield compared to that obtained by cell culture (diagnostic yields, 60% and 35.5%, respectively). In conclusion, the RespiFinder assay provides a user-friendly and high-throughput tool for the simultaneous detection of 15 respiratory viruses with excellent overall performance statistics.