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. 2018 Mar 7;7(1):18.
doi: 10.1038/s41426-017-0016-7.

Rapid detection of MERS coronavirus-like viruses in bats: pote1ntial for tracking MERS coronavirus transmission and animal origin

Patrick C Y Woo  1   2   3   4 , Susanna K P Lau  5   6   7   8 , Yixin Chen  9 , Emily Y M Wong  10 , Kwok-Hung Chan  10 , Honglin Chen  11   12   13   10 , Libiao Zhang  14 , Ningshao Xia  9 , Kwok-Yung Yuen  11   12   13   10
Affiliations

Rapid detection of MERS coronavirus-like viruses in bats: pote1ntial for tracking MERS coronavirus transmission and animal origin

Patrick C Y Woo et al. Emerg Microbes Infect. .

Abstract

Recently, we developed a monoclonal antibody-based rapid nucleocapsid protein detection assay for diagnosis of MERS coronavirus (MERS-CoV) in humans and dromedary camels. In this study, we examined the usefulness of this assay to detect other lineage C betacoronaviruses closely related to MERS-CoV in bats. The rapid MERS-CoV nucleocapsid protein detection assay was tested positive in 24 (88.9%) of 27 Tylonycteris bat CoV HKU4 (Ty-BatCoV-HKU4) RNA-positive alimentary samples of Tylonycteris pachypus and 4 (19.0%) of 21 Pipistrellus bat CoV HKU5 (Pi-BatCoV-HKU5) RNA-positive alimentary samples of Pipistrellus abramus. There was significantly more Ty-BatCoV-HKU4 RNA-positive alimentary samples than Pi-BatCoV-HKU5 RNA-positive alimentary samples that were tested positive by the rapid MERS-CoV nucleocapsid protein detection assay (P < 0.001 by Chi-square test). The rapid assay was tested negative in all 51 alimentary samples RNA-positive for alphacoronaviruses (Rhinolophus bat CoV HKU2, Myotis bat CoV HKU6, Miniopterus bat CoV HKU8 and Hipposideros batCoV HKU10) and 32 alimentary samples positive for lineage B (SARS-related Rhinolophus bat CoV HKU3) and lineage D (Rousettus bat CoV HKU9) betacoronaviruses. No significant difference was observed between the viral loads of Ty-BatCoV-HKU4/Pi-BatCoV-HKU5 RNA-positive alimentary samples that were tested positive and negative by the rapid test (Mann-Witney U test). The rapid MERS-CoV nucleocapsid protein detection assay is able to rapidly detect lineage C betacoronaviruses in bats. It detected significantly more Ty-BatCoV-HKU4 than Pi-BatCoV-HKU5 because MERS-CoV is more closely related to Ty-BatCoV-HKU4 than Pi-BatCoV-HKU5. This assay will facilitate rapid on-site mass screening of animal samples for ancestors of MERS-CoV and tracking transmission in the related bat species.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1. Rapid MERS-CoV nucleocapsid protein detection assay showing.
a positive, weakly positive and negative results for selected Ty-BatCoV-HKU4 positive and Pi-BatCoV-HKU5 positive samples and negative results for Tylonycteris pachypus and Pipistrellus abramus CoV RNA-negative samples; and (b) results for serial dilutions of recombinant nucleocapsid protein of Ty-BatCoV-HKU4 strain SM2A
Fig. 2
Fig. 2
Phylogenetic analysis of (a) nucleocapsid, (b) spike and (c) RdRp protein of Ty-BatCoV-HKU4, Pi-BatCoV-HKU5, and MERS-CoV. The trees were constructed by maximum-likelihood method in MEGA 6 using substitution model WAG+G (nucleocapsid), WAG+F+I+G (spike) and LG+G (RdRp) with SARS-CoV as the outgroup. The percentage of trees in which the associated taxa clustered together next to the branches with bootstrap values was calculated from 1000 trees. The scale bars indicate the number of substitutions per 10, 5, and 20 amino acids, respectively
Fig. 3
Fig. 3. Multiple alignment of the amino acid sequences of the nucleocapsid proteins of MERS-CoV, Ty-BatCoV-HKU4, and Pi-BatCoV-HKU5.
Red color shows the amino acid residues that were identical between the nucleocapsid proteins of MERS-CoV and Ty-BatCoV-HKU4 but not with that of Pi-BatCoV-HKU5. Blue color shows the amino acid residues that were identical between nucleocapsid proteins of MERS-CoV and Pi-BatCoV-HKU5 but not with that of Ty-BatCoV-HKU4
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