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Nucleic Acid Visualization Assay for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) by Targeting the UpE and N Gene
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  • Pei Huang,
  • Hongli Jin,
  • Yongkun Zhao,
  • Entao Li,
  • Feihu Yan,
  • Hang Chi,
  • Qi Wang,
  • Qiuxue Han,
  • Ruo Mo,
  • Yumeng Song,
  • Jinhao Bi,
  • Cui Jiao,
  • Wujian Li,
  • Hongbin He,
  • Hong-Mei Wang,
  • Aimin Ma,
  • Na Feng,
  • Jianzhong Wang,
  • Tiecheng Wang,
  • Songtao Yang,
  • Yuwei Gao,
  • Xianzhu Xia,
  • Hualei Wang
Pei Huang
Jilin Agricultural University
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Hongli Jin
Jilin University
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Yongkun Zhao
Academy of Military Medical Sciences
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Entao Li
Academy of Military Medical Sciences
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Feihu Yan
Academy of Military Medical Sciences
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Hang Chi
Academy of Military Medical Sciences
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Qi Wang
Academy of Military Medical Sciences
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Qiuxue Han
Academy of Military Medical Sciences
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Ruo Mo
Jilin Agricultural University
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Yumeng Song
Jilin University
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Jinhao Bi
Jilin Agricultural University
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Cui Jiao
Jilin University
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Wujian Li
Jilin University
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Hongbin He
Shandong Normal University
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Hong-Mei Wang
Shandong Normal University
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Aimin Ma
Changchun Medical College
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Na Feng
Academy of Military Medical Sciences
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Jianzhong Wang
Jilin Agricultural University
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Tiecheng Wang
Academy of Military Medical Sciences
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Songtao Yang
Academy of Military Medical Sciences
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Yuwei Gao
Academy of Military Medical Sciences
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Xianzhu Xia
Academy of Military Medical Sciences
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Hualei Wang
Jilin University
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Abstract

Since its first emergence in 2012, cases of infection with Middle East respiratory syndrome coronavirus (MERS-CoV) have continued to occur. In this study, we present two nucleic acid visualization assays that target the MERS-CoV UpE and N genes as a panel that combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). The limit of detection was 1.2×10^1 copies/μl for the UpE assay and 1.2 copies/μl for the N assay. The two assays exhibited no cross-reactivity with multiple CoVs, including the bat severe acute respiratory syndrome related coronavirus (SARSr-CoV), the bat coronavirus HKU4, and the human coronaviruses 229E, OC43, HKU1 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The RT-RPA-VF assay does not require sophisticated equipment and provides rapid detection within 30 min, so it has potential for use in surveillance and detection of MERS-CoV in low-resource settings.