Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses.
Development of three multiplex RT-PCR assays for the detection of 12 respiratory RNA viruses.
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DOI：
10.1016/j.jviromet.2005.01.020
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发表时间：
2005-06
影响因子：
3.1
通讯作者：
Freymuth F
中科院分区：
文献类型：
--
作者：
Bellau-Pujol S;Vabret A;Legrand L;Dina J;Gouarin S;Petitjean-Lecherbonnier J;Pozzetto B;Ginevra C;Freymuth F
建立了3种多重半巢式RT-PCR方法，可同时检测流感病毒A、B、C、人呼吸道合胞病毒（hRSV）、人偏肺病毒（hMPV）、副流感病毒1-4型（PIV-1、PIV-2、PIV-3、PIV-4）、人冠状病毒OC 43、229 E（HCoV）和鼻病毒（hRV）等12种呼吸道病毒RNA。在其中一项RT-PCR试验中包括内部扩增对照。多重RT-PCR 1和半巢式多重RT-PCR 1分别检测到1和0.1 TCID 50的RSV A，以及0.01和0.001 TCID 50的流感病毒A/H3 N2。对203例住院儿童的鼻吸出物进行了回顾性检测，并与两种常规方法进行了比较：直接免疫荧光检测和病毒分离技术。几乎所有免疫荧光法和/或病毒分离技术检测阳性的样本（89/91）都被多重检测法检测到。该方法还检测到另外85种病毒和33种合并感染。这些多重半巢式RT-PCR检测的总体灵敏度（98%）、快速性和增强的效率表明，与用于检测广谱呼吸道病毒的常规方法相比，它们将是一个显著的改进。
Three multiplex hemi-nested RT-PCR assays were developed to detect simultaneously 12 RNA respiratory viruses: influenza viruses A, B and C, human respiratory syncytial virus (hRSV), human metapneumovirus (hMPV), parainfluenza virus types 1–4 (PIV-1, -2, -3 and -4), human coronavirus OC43 and 229E (HCoV) and rhinovirus (hRV). An internal amplification control was included in one of the RT-PCR assays. The RT-PCR multiplex 1 and the hemi-nested multiplex 1 detected 1 and 0.1 TCID50 of RSV A, respectively, and 0.01 and 0.001 TCID50 of influenza virus A/H3N2, respectively. Two hundred and three nasal aspirates from hospitalised children were retrospectively tested in comparison with two conventional methods: direct immunofluorescence assay and viral isolation technique. Almost all samples (89/91) that were positive by immunofluorescence assay and/or viral isolation technique were detected by the multiplex assay. This method also detected an additional 85 viruses and 33 co-infections. The overall sensitivity (98%), rapidity and enhanced efficiency of these multiplex hemi-nested RT-PCR assays suggest that they would be a significant improvement over conventional methods for the detection of a broad spectrum of respiratory viruses.
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影响因子：
3.8
作者：
CANE, PA;PRINGLE, CR
通讯作者：
PRINGLE, CR
影响因子：
9.4
作者：
Aguilar, JC;Pérez-Breña, MP;Echevarria, JE
通讯作者：
Echevarria, JE
影响因子：
8.8
作者：
Casiano-Colón, AE;Hulbert, BB;Falsey, AR
通讯作者：
Falsey, AR
影响因子：
12.7
作者：
Coiras, MT;Aguilar, JC;Pérez-Breña, P
通讯作者：
Pérez-Breña, P
影响因子：
9.4
作者：
Dingle, KE;Crook, D;Jeffery, K
通讯作者：
Jeffery, K