Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays
Simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-PCR assays
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DOI：
10.1002/jmv.20008
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发表时间：
2004-03-01
影响因子：
12.7
通讯作者：
Pérez-Breña, P
中科院分区：
文献类型：
--
作者：
Coiras, MT;Aguilar, JC;Pérez-Breña, P
有必要对儿童、老年人和免疫功能低下患者的下呼吸道感染进行快速、敏感和准确的诊断，这些患者容易出现严重的并发症。多重RT-巢式聚合酶链式反应因其高度的特异性和敏感性而被广泛应用于传染病无关病毒的同时检测。本文介绍了一种新的多重RT-PCR方法。这种方法包括针对人类副流感病毒血凝素、人类冠状病毒刺突蛋白以及人类肠道病毒和鼻病毒多蛋白基因区域的嵌套引物集。它允许快速、灵敏和同时检测和分型四种类型的副流感病毒(1、2、3、4AB)、人类冠状病毒229E和OC43，以及对肠道病毒和鼻病毒的通用检测。用这种多重检测方法与我们小组以前描述的另一种方法一起对201份临床标本进行检测，以同时检测和分型流感病毒、呼吸道合胞病毒和所有血清型的腺病毒，涵盖了对大多数引起人类呼吸道传染病的病毒的检测。将获得的结果与之前描述的所有人类副流感病毒类型的常规病毒培养、免疫荧光检测和第三次多重RT-PCR检测进行比较。综上所述，这两种多重RT-PCR方法都提供了一个能够同时检测和鉴定临床标本中14种不同呼吸道病毒的系统，具有很高的敏感性和特异性，对于人群中这些病毒的常规诊断和调查是有用的。(C)2004年Wiley-Liss公司
There is a need for rapid, sensitive, and accurate diagnosis of lower respiratory tract infections in children, elderly, and immunocompromised patients, who are susceptible to serious complications. The multiplex RT-nested PCR assay has been used widely for simultaneous detection of non-related viruses involved in infectious diseases because of its high specificity and sensitivity. A new multiplex RT-PCR assay is described in this report. This approach includes nested primer sets targeted to conserve regions of human parainfluenza virus haemagglutinin, human coronavirus spike protein, and human enterovirus and rhinovirus polyprotein genes. It permits rapid, sensitive, and simultaneous detection and typing of the four types of parainfluenza viruses (1, 2, 3, 4AB), human coronavirus 229E and OC43, and the generic detection of enteroviruses and rhinoviruses. The testing of 201 clinical specimens with this multiplex assay along with other one formerly described by our group to simultaneously detect and type the influenza viruses, respiratory syncytial viruses, and a generic detection of all serotypes of adenovirus, covers the detection of most viruses causing respiratory infectious disease in humans. The results obtained were compared with conventional viral culture, immunofluorescence assay, and a third multiplex RT-PCR assay for all human parainfluenza viruses types described previously. In conclusion, both multiplex RT-PCR assays provide a system capable of detecting and identifying simultaneously 14 different respiratory viruses in clinical specimens with high sensitivity and specificity, being useful for routine diagnosis and survey of these viruses within the population. (C) 2004 Wiley-Liss, Inc.