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Emerging Microbes & Infections Volume 7, 2018 - Issue 1
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Original Articles

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

Patrick C. Y. WooState Key Laboratory of Emerging Infectious DiseasesThe University of Hong KongHong KongChina;Research Centre of Infection and ImmunologyThe University of Hong KongHong KongChina;Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesThe University of Hong KongHong KongChina;Department of MicrobiologyThe University of Hong KongHong KongChinaCorrespondence[email protected]
,
Susanna K. P. LauState Key Laboratory of Emerging Infectious DiseasesThe University of Hong KongHong KongChina;Research Centre of Infection and ImmunologyThe University of Hong KongHong KongChina;Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesThe University of Hong KongHong KongChina;Department of MicrobiologyThe University of Hong KongHong KongChinaCorrespondence[email protected]
,
Yixin ChenState Key Laboratory of Molecular Vaccinology and Molecular Diagnostics & National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, School of Public HealthXiamen University, Xiamen361102FujianChina
,
Emily Y. M. WongDepartment of MicrobiologyThe University of Hong KongHong KongChina
,
Kwok-Hung ChanDepartment of MicrobiologyThe University of Hong KongHong KongChina
,
Honglin ChenState Key Laboratory of Emerging Infectious DiseasesThe University of Hong KongHong KongChina;Research Centre of Infection and ImmunologyThe University of Hong KongHong KongChina;Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesThe University of Hong KongHong KongChina;Department of MicrobiologyThe University of Hong KongHong KongChinaORCID Iconhttp://orcid.org/0000-0001-5108-8338
ORCID Icon,
Libiao ZhangGuangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and UtilizationGuangdong Institute of Applied Biological ResourcesGuangzhouChina
,
Ningshao XiaState Key Laboratory of Molecular Vaccinology and Molecular Diagnostics & National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, School of Public HealthXiamen University, Xiamen361102FujianChina
&
Kwok-Yung YuenState Key Laboratory of Emerging Infectious DiseasesThe University of Hong KongHong KongChina;Research Centre of Infection and ImmunologyThe University of Hong KongHong KongChina;Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseasesThe University of Hong KongHong KongChina;Department of MicrobiologyThe University of Hong KongHong KongChina
 show all
Pages 1-7 | Received 14 Jul 2017, Accepted 14 Dec 2017, Published online: 07 Mar 2018

Figures & data

Alimentary samples tested in the present study

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. 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

Rapid test results and viral loads of Ty-BatCoV-HKU4 and Pi-BatCoV-HKU5 positive samples

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. 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 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

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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