Performance and clinical validation of the RealStar® MERS-CoV Kit for detection of Middle East respiratory syndrome coronavirus RNA
Performance and clinical validation of the RealStar® MERS-CoV Kit for detection of Middle East respiratory syndrome coronavirus RNA
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DOI：
10.1016/j.jcv.2014.03.012
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发表时间：
2014-06-01
影响因子：
8.8
通讯作者：
Drosten, Christian
中科院分区：
文献类型：
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作者：
Corman, Victor Max;Oelschlaeger, Stephan;Drosten, Christian
背景：2012年，中东地区出现了一种引起呼吸道疾病的高致病性人类冠状病毒。这种病毒的内部分子诊断方法称为中东呼吸综合征冠状病毒（MERS-CoV），可以在患者样本中进行灵敏的MERS-CoV RNA检测。快速诊断是重要的，以管理人类病例和跟踪可能的contacts.Objectives：本研究的目的是提高现有的核酸amplificationbased诊断方法的可用性MERS冠状病毒infections通过提供实时RT-PCR试剂盒，包括内部控制和两个目标区域推荐的世界卫生组织（WHO）。为了验证该试剂盒（RealStar（R）MERS-CoV RT-PCR试剂盒1.0，Altona Diagnostics GmbH，Hamburg，德国），使用来自慕尼黑的一个MERS-CoV病例的临床样本和患有其他呼吸系统疾病的患者的呼吸道样本。基于这些检测，一个即用型实时RT-PCR试剂盒，具有upE和ORF 1A检测开发，验证和比较，以建立在内部versions.Results：包括在试剂盒中的两个RT-PCR检测的性能是可比的内部检测。它们显示出高的分析灵敏度（upE：5.3拷贝/反应; ORF 1A：9.3拷贝/反应），与其他呼吸道病原体无交叉反应性，并在患者样本中检测到MERS-CoV RNA，其检测方式与内部版本几乎相同。结论：该试剂盒是一种有价值的工具，可用于辅助MERS-CoV疑似病例的快速诊断、患者管理和流行病学研究。(C)2014爱思唯尔有限公司版权所有。
Background: A highly pathogenic human coronavirus causing respiratory disease emerged in the Middle East region in 2012. In-house molecular diagnostic methods for this virus termed Middle East respiratory syndrome coronavirus (MERS-CoV) allowed sensitive MERS-CoV RNA detection in patient samples. Fast diagnosis is important to manage human cases and trace possible contacts.Objectives: The aim of this study was to improve the availability of existing nucleic acid amplificationbased diagnostic methods for MERS-CoV infections by providing a real-time RT-PCR kit, including an internal control and two target regions recommended by the World Health Organization (WHO). And to validate this kit (RealStar (R) MERS-CoV RT-PCR kit 1.0, Altona Diagnostics GmbH, Hamburg, Germany) using clinical samples of one MERS-CoV case from Munich and respiratory samples of patients with other respiratory diseases.Study design: An internal amplification control was included into the RT-PCR assays targeting the genomic region upstream of the Envelope gene (upE) and within open reading frame (ORF) 1A. Based on these assays, a ready-to-use real-time RT-PCR kit featuring both the upE and ORF1A assays was developed, validated and compared to the established in-house versions.Results: The performance of both RT-PCR assays included in the kit is comparable to the in-house assays. They show high analytical sensitivity (upE: 5.3 copies/reaction; ORF1A: 9.3 copies/reaction), no crossreactivity with other respiratory pathogens and detected MERS-CoV RNA in patient samples in almost the same manner as the in-house versions.Conclusion: The kit is a valuable tool for assisting in the rapid diagnosis, patient management and epidemiology of suspected MERS-CoV cases. (C) 2014 Elsevier B.V. All rights reserved.