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Detection and full genome characterization of two beta CoV viruses related to Middle East respiratory syndrome from bats in Italy.
Virol J. 2017 12 19; 14(1):239.VJ

Abstract

BACKGROUND

Middle East respiratory syndrome coronavirus (MERS-CoV), which belongs to beta group of coronavirus, can infect multiple host species and causes severe diseases in humans. Multiple surveillance and phylogenetic studies suggest a bat origin. In this study, we describe the detection and full genome characterization of two CoVs closely related to MERS-CoV from two Italian bats, Pipistrellus kuhlii and Hypsugo savii.

METHODS

Pool of viscera were tested by a pan-coronavirus RT-PCR. Virus isolation was attempted by inoculation in different cell lines. Full genome sequencing was performed using the Ion Torrent platform and phylogenetic trees were performed using IQtree software. Similarity plots of CoV clade c genomes were generated by using SSE v1.2. The three dimensional macromolecular structure (3DMMS) of the receptor binding domain (RBD) in the S protein was predicted by sequence-homology method using the protein data bank (PDB).

RESULTS

Both samples resulted positive to the pan-coronavirus RT-PCR (IT-batCoVs) and their genome organization showed identical pattern of MERS CoV. Phylogenetic analysis showed a monophyletic group placed in the Beta2c clade formed by MERS-CoV sequences originating from humans and camels and bat-related sequences from Africa, Italy and China. The comparison of the secondary and 3DMMS of the RBD of IT-batCoVs with MERS, HKU4 and HKU5 bat sequences showed two aa deletions located in a region corresponding to the external subdomain of MERS-RBD in IT-batCoV and HKU5 RBDs.

CONCLUSIONS

This study reported two beta CoVs closely related to MERS that were obtained from two bats belonging to two commonly recorded species in Italy (P. kuhlii and H. savii). The analysis of the RBD showed similar structure in IT-batCoVs and HKU5 respect to HKU4 sequences. Since the RBD domain of HKU4 but not HKU5 can bind to the human DPP4 receptor for MERS-CoV, it is possible to suggest also for IT-batCoVs the absence of DPP4-binding potential. More surveillance studies are needed to better investigate the potential intermediate hosts that may play a role in the interspecies transmission of known and currently unknown coronaviruses with particular attention to the S protein and the receptor specificity and binding affinity.

Authors+Show Affiliations

Department of Virology, Istituto Zooprofilattico Sperimentale Lombardia ed Emilia Romagna, Via Antonio Bianchi 9, 25124, Brescia, Italy. anamaria.morenomartin@izsler.it.Department of Virology, Istituto Zooprofilattico Sperimentale Lombardia ed Emilia Romagna, Via Antonio Bianchi 9, 25124, Brescia, Italy.Department of Food Safety, Istituto Superiore di Sanità. Nutrition and Veterinary Public Health, Viale Regina Elena 299, 00161, Rome, Italy. Dept. of Sciences, University Roma Tre, Viale Guglielmo Marconi 446, 00146, Rome, Italy.Department of Food Safety, Istituto Superiore di Sanità. Nutrition and Veterinary Public Health, Viale Regina Elena 299, 00161, Rome, Italy.Department of Food Safety, Istituto Superiore di Sanità. Nutrition and Veterinary Public Health, Viale Regina Elena 299, 00161, Rome, Italy.Department of Virology, Istituto Zooprofilattico Sperimentale Lombardia ed Emilia Romagna, Via Antonio Bianchi 9, 25124, Brescia, Italy.Department of Virology, Istituto Zooprofilattico Sperimentale Lombardia ed Emilia Romagna, Via Antonio Bianchi 9, 25124, Brescia, Italy.Department of Virology, Istituto Zooprofilattico Sperimentale Lombardia ed Emilia Romagna, Via Antonio Bianchi 9, 25124, Brescia, Italy.University Campus Bio-Medico of Rome, Via Álvaro del Portillo, 21, 00128, Rome, Italy.Department of Food Safety, Istituto Superiore di Sanità. Nutrition and Veterinary Public Health, Viale Regina Elena 299, 00161, Rome, Italy.University Campus Bio-Medico of Rome, Via Álvaro del Portillo, 21, 00128, Rome, Italy.Department of Food Safety, Istituto Superiore di Sanità. Nutrition and Veterinary Public Health, Viale Regina Elena 299, 00161, Rome, Italy.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29258555

Citation

Moreno, Ana, et al. "Detection and Full Genome Characterization of Two Beta CoV Viruses Related to Middle East Respiratory Syndrome From Bats in Italy." Virology Journal, vol. 14, no. 1, 2017, p. 239.
Moreno A, Lelli D, de Sabato L, et al. Detection and full genome characterization of two beta CoV viruses related to Middle East respiratory syndrome from bats in Italy. Virol J. 2017;14(1):239.
Moreno, A., Lelli, D., de Sabato, L., Zaccaria, G., Boni, A., Sozzi, E., Prosperi, A., Lavazza, A., Cella, E., Castrucci, M. R., Ciccozzi, M., & Vaccari, G. (2017). Detection and full genome characterization of two beta CoV viruses related to Middle East respiratory syndrome from bats in Italy. Virology Journal, 14(1), 239. https://doi.org/10.1186/s12985-017-0907-1
Moreno A, et al. Detection and Full Genome Characterization of Two Beta CoV Viruses Related to Middle East Respiratory Syndrome From Bats in Italy. Virol J. 2017 12 19;14(1):239. PubMed PMID: 29258555.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection and full genome characterization of two beta CoV viruses related to Middle East respiratory syndrome from bats in Italy. AU - Moreno,Ana, AU - Lelli,Davide, AU - de Sabato,Luca, AU - Zaccaria,Guendalina, AU - Boni,Arianna, AU - Sozzi,Enrica, AU - Prosperi,Alice, AU - Lavazza,Antonio, AU - Cella,Eleonora, AU - Castrucci,Maria Rita, AU - Ciccozzi,Massimo, AU - Vaccari,Gabriele, Y1 - 2017/12/19/ PY - 2017/10/20/received PY - 2017/12/06/accepted PY - 2017/12/21/entrez PY - 2017/12/21/pubmed PY - 2018/7/18/medline KW - Bats KW - Full genome sequencing KW - Italy KW - MERS-like Beta-CoV viruses KW - Phylogenetic and molecular analyses SP - 239 EP - 239 JF - Virology journal JO - Virol J VL - 14 IS - 1 N2 - BACKGROUND: Middle East respiratory syndrome coronavirus (MERS-CoV), which belongs to beta group of coronavirus, can infect multiple host species and causes severe diseases in humans. Multiple surveillance and phylogenetic studies suggest a bat origin. In this study, we describe the detection and full genome characterization of two CoVs closely related to MERS-CoV from two Italian bats, Pipistrellus kuhlii and Hypsugo savii. METHODS: Pool of viscera were tested by a pan-coronavirus RT-PCR. Virus isolation was attempted by inoculation in different cell lines. Full genome sequencing was performed using the Ion Torrent platform and phylogenetic trees were performed using IQtree software. Similarity plots of CoV clade c genomes were generated by using SSE v1.2. The three dimensional macromolecular structure (3DMMS) of the receptor binding domain (RBD) in the S protein was predicted by sequence-homology method using the protein data bank (PDB). RESULTS: Both samples resulted positive to the pan-coronavirus RT-PCR (IT-batCoVs) and their genome organization showed identical pattern of MERS CoV. Phylogenetic analysis showed a monophyletic group placed in the Beta2c clade formed by MERS-CoV sequences originating from humans and camels and bat-related sequences from Africa, Italy and China. The comparison of the secondary and 3DMMS of the RBD of IT-batCoVs with MERS, HKU4 and HKU5 bat sequences showed two aa deletions located in a region corresponding to the external subdomain of MERS-RBD in IT-batCoV and HKU5 RBDs. CONCLUSIONS: This study reported two beta CoVs closely related to MERS that were obtained from two bats belonging to two commonly recorded species in Italy (P. kuhlii and H. savii). The analysis of the RBD showed similar structure in IT-batCoVs and HKU5 respect to HKU4 sequences. Since the RBD domain of HKU4 but not HKU5 can bind to the human DPP4 receptor for MERS-CoV, it is possible to suggest also for IT-batCoVs the absence of DPP4-binding potential. More surveillance studies are needed to better investigate the potential intermediate hosts that may play a role in the interspecies transmission of known and currently unknown coronaviruses with particular attention to the S protein and the receptor specificity and binding affinity. SN - 1743-422X UR - https://www.unboundmedicine.com/medline/citation/29258555/Detection_and_full_genome_characterization_of_two_beta_CoV_viruses_related_to_Middle_East_respiratory_syndrome_from_bats_in_Italy_ L2 - https://virologyj.biomedcentral.com/articles/10.1186/s12985-017-0907-1 DB - PRIME DP - Unbound Medicine ER -