J Korean Med Sci.  2016 Feb;31(2):315-320. 10.3346/jkms.2016.31.2.315.

Isolation of Middle East Respiratory Syndrome Coronavirus from a Patient of the 2015 Korean Outbreak

• Park WB
• Kwon NJ
• Choe PG
• Choi SJ
• Oh HS
• Lee SM
• Chong H
• Kim JI
• Song KH
• Bang JH
• Kim ES
• Kim HB
• Park SW
• Kim NJ
• Oh MD

• KMID: 2360059
• DOI: http://doi.org/10.3346/jkms.2016.31.2.315

Abstract

During the 2015 outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV) in Korea, 186 persons were infected, resulting in 38 fatalities. We isolated MERS-CoV from the oropharyngeal sample obtained from a patient of the outbreak. Cytopathic effects showing detachment and rounding of cells were observed in Vero cell cultures 3 days after inoculation of the sample. Spherical virus particles were observed by transmission electron microscopy. Full-length genome sequence of the virus isolate was obtained and phylogenetic analyses showed that it clustered with clade B of MERS-CoV.

Keyword

Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Microscopy, Electron; Phylogeny; Korea

MeSH Terms

Animals
Cercopithecus aethiops
Coronavirus Infections/*diagnosis/epidemiology/*virology
Disease Outbreaks
Humans
Microscopy, Electron
Middle East Respiratory Syndrome Coronavirus/classification/genetics/*isolation & purification/ultrastructure
Phylogeny
Polymerase Chain Reaction
RNA, Viral/analysis/chemistry/metabolism
Republic of Korea/epidemiology
Sequence Analysis, RNA
Vero Cells
RNA, Viral

Figure

• 

  Fig. 1 Cytopathic effects of MERS-CoV in Vero cell cultures and Electron microscopy image of MERS-CoV. Vero cells were inoculated with oropharyngeal swab sample. (A) Vero cell cultures in negative control. (B) Cytopathic effects (rounding and detachment of cells) in Vero cell cultures 3 days after inoculation of the sample. (C, D) Transmission electron microscopy image of Vero cells infected with MERS-CoV. White arrow denotes nuclear membrane. Brown scale bar indicates 100 μm (A and B). Black scale bar indicates 500 nm (C) and white scale bar does 200 nm (D).

• 

  Fig. 2 Molecular phylogenetic analysis. Phylogenetic tree on complete genome (A), S genes (B), and ORF1ab genes (C) for the 101 gene sequences of MERS-CoV. The evolutionary history was inferred by using the maximum likelihood method based on the Tamura-Nei model (16). Evolutionary analyses were conducted in MEGA6 (17). Red box indicates our virus isolate.

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