...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>BMC Pediatrics >Simultaneous detection of respiratory syncytial virus and human metapneumovirus by one-step multiplex real-time RT-PCR in patients with respiratory symptoms
【24h】

Simultaneous detection of respiratory syncytial virus and human metapneumovirus by one-step multiplex real-time RT-PCR in patients with respiratory symptoms

机译：一站式多重实时RT-PCR同时检测呼吸道症状患者的呼吸道合胞病毒和人类偏肺病毒

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Background Both respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) are important viral pathogens causing respiratory tract infection (RTI) in the pediatric population. However, the clinical manifestations of RSV and hMPV infections are similar. Therefore, a reliable and rapid diagnostic tool is needed for diagnostic performance. Methods In order to optimize diagnosis efficiency of RTI, the aim of this study is to establish a rapid and advanced method for simultaneous detecting RSV and hMPV in nasopharyngeal aspirates specimens from patients. We designed a one-step triplex real-time RT-PCR (qRT-PCR) protocol using TaqMan probes for detecting RSV and hMPV. The plasmid clones containing RSV nucleoprotein gene and hMPV fusion gene were established as reference standards. We used virus culture supernatants from 86 known pediatric RTI patient to test the specificity and sensitivity of our assay. Then we used total 222 nasopharyngeal aspirates specimens from pediatric patients hospitalized with respiratory symptoms to evaluate our assay. Results Our one-step triplex qRT-PCR assay showed 100% sensitivity and specificity in testing RSV and hMPV in 86 known virus culture supernatants, with excellent linearity (R2?>?0.99) and reliable reproducibility (CV lower than 1.04%). This assay has a wide dynamic range 102-109copies/reaction (limit of detection; LOD?=?100 copies/reaction). A total of 222 patients hospitalized with respiratory symptoms were enrolled for clinical evaluation. In these samples, our qRT-PCR assay detected 68 RSV positive and 18 hMPV positive cases. However, standard virus culture only detected 8 RSV positive cases and 0 hMPV cases. Based on this improved triplex qRT-PCR assay, we found that RSV infection was associated with severe inflammation by chest X-ray and occurrence of pneumonia which were not observed previously. Conclusions In summary, we have developed a highly specific and sensitive one-step triplex qRT-PCR assay to detect hMPV and RSV simultaneously. This assay offers a valuable tool for routine diagnosis.
机译：背景技术呼吸道合胞病毒（RSV）和人间质肺病毒（hMPV）都是引起儿科人群呼吸道感染（RTI）的重要病毒病原体。但是，RSV和hMPV感染的临床表现相似。因此，需要可靠且快速的诊断工具来提高诊断性能。方法为了优化RTI的诊断效率，本研究的目的是建立一种快速，先进的方法，用于同时检测患者鼻咽抽吸物中的RSV和hMPV。我们使用TaqMan探针设计了一步一步三重实时RT-PCR（qRT-PCR）协议，用于检测RSV和hMPV。建立含有RSV核蛋白基因和hMPV融合基因的质粒克隆作为参考标准。我们使用来自86名已知RTI患儿的病毒培养上清液来测试我们测定的特异性和敏感性。然后，我们使用来自呼吸道症状住院的小儿患者的总共222例鼻咽抽吸物来评估我们的测定。结果我们的一步三重qRT-PCR分析显示出在86种已知病毒培养上清液中检测RSV和hMPV的敏感性和特异性为100％，具有出色的线性（R 2 →> 0.99）和可靠的重现性（ CV低于1.04％）。该测定法具有10 2 -10 9 拷贝/反应的宽动态范围（检测限;LOD≥100拷贝/反应）。共有222例因呼吸道症状住院的患者入选临床评估。在这些样品中，我们的qRT-PCR分析检测到68例RSV阳性和18例hMPV阳性。但是，标准病毒培养仅检测到8例RSV阳性病例和0例hMPV病例。基于这种改进的三重qRT-PCR分析，我们发现RSV感染与胸部X射线引起的严重炎症和肺炎的发生有关，这是以前未曾观察到的。结论总之，我们已经开发了一种高度特异性和灵敏的一步三重qRT-PCR检测方法，可同时检测hMPV和RSV。该测定法为常规诊断提供了有价值的工具。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号