A Rapid and Specific Assay for the Detection of MERS-CoV

A Rapid and Specific Assay for the Detection of MERS-CoV

Pei Huang, Hualei Wang, Zengguo Cao, Hong-li Jin, Hang Chi, Jincun Zhao, Beibei Yu, Feihu Yan, Xingxing Hu, Fangfang Wu, Cuicui Jiao, Pengfei Hou, Shengnan Xu, Yongkun Zhao, Na Feng, Jian-Zhong Wang, Weiyang Sun, Tiecheng Wang, Yuwei Gao, Songtao Yang, Xianzhu Xia

Middle East respiratory syndrome coronavirus (MERS-CoV) is a novel human coronavirus that can cause human respiratory disease. The development of a detection method for this virus that can lead to rapid and accurate diagnosis would be significant. In this study, we established a nucleic acid visualization technique that combines the reverse transcription loop-mediated isothermal amplification technique and a vertical flow visualization strip (RT-LAMP-VF) to detect the N gene of MERS-CoV. The RT-LAMP-VF assay was performed in a constant temperature water bath for 30 min, and the result was visible by the naked eye within 5 min. The RT-LAMP-VF assay was capable of detecting 2 × 101 copies/μl of synthesized RNA transcript and 1 × 101 copies/μl of MERS-CoV RNA. The method exhibits no cross-reactivities with multiple CoVs including SARS-related (SARSr)-CoV, HKU4, HKU1, OC43 and 229E, and thus exhibits high specificity. Compared to the real-time RT-PCR (rRT-PCR) method recommended by the World Health Organization (WHO), the RT-LAMP-VF assay is easy to handle, does not require expensive equipment and can rapidly complete detection within 35 min.

2018

Middle East respiratory syndrome coronavirus, reverse transcription loop-mediated isothermal amplification, nucleic acid visualization, visual detection, RT-LAMP-VF

DOI: 10.3389/fmicb.2018.01101

Frontiers in Microbiology

Frontiers Media S.A.

Microbiology

EN