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 amplification-based 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 ® 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 cross-reactivity 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.
【저자키워드】 coronavirus, MERS-CoV, PCR, diagnostic assay, Middle East respiratory syndrome, 【초록키워드】 Epidemiology, Diagnosis, virus, respiratory diseases, cross-reactivity, amplification, nucleic acid, Internal control, Germany, management, Respiratory disease, Patient, envelope, WHO, molecular, real-time RT-PCR, RT-PCR assay, Diagnostic method, analytical sensitivity, Munich, Middle East, World Health Organization, Frame, genomic region, respiratory pathogen, respiratory syndrome coronavirus, MERS-CoV infection, RT-PCR kit, Hamburg, upstream, highly pathogenic, objective, respiratory sample, IMPROVE, Altona Diagnostics, clinical sample, Result, assays, ORF, comparable, target region, MERS-CoV RNA, 【제목키워드】 RNA, performance, Middle East, respiratory syndrome coronavirus,