#Isolation of #SARS-CoV-2-related #coronavirus from #Malayan #pangolins (Nature, abstract)

[Source: Nature, full page: (LINK). Abstract, edited.]

This is an unedited manuscript that has been accepted for publication. Nature Research are providing this early version of the manuscript as a service to our authors and readers. The manuscript will undergo copyediting, typesetting and a proof review before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers apply.

Isolation of SARS-CoV-2-related coronavirus from Malayan pangolins

Kangpeng Xiao, Junqiong Zhai, Yaoyu Feng, Niu Zhou, Xu Zhang, Jie-Jian Zou, Na Li, Yaqiong Guo, Xiaobing Li, Xuejuan Shen, Zhipeng Zhang, Fanfan Shu, Wanyi Huang, Yu Li, Ziding Zhang, Rui-Ai Chen, Ya-Jiang Wu, Shi-Ming Peng, Mian Huang, Wei-Jun Xie, Qin-Hui Cai, Fang-Hui Hou, Wu Chen, Lihua Xiao & Yongyi Shen

Nature (2020)

 

Abstract

The outbreak of COVID-19 poses unprecedent challenges to global health1. The new coronavirus, SARS-CoV-2, shares high sequence identity to SARS-CoV and a bat coronavirus RaTG132. While bats may be the reservoir host for various coronaviruses3,4, whether SARS-CoV-2 has other hosts remains ambiguous. In this study, one coronavirus isolated from a Malayan pangolin showed 100%, 98.6%, 97.8% and 90.7% amino acid identity with SARS-CoV-2 in the E, M, N and S genes, respectively. In particular, the receptor-binding domain within the S protein of the Pangolin-CoV is virtually identical to that of SARS-CoV-2, with one noncritical amino acid difference. Results of comparative genomic analysis suggest that SARS-CoV-2 might have originated from the recombination of a Pangolin-CoV-like virus with a Bat-CoV-RaTG13-like virus. The Pangolin-CoV was detected in 17 of 25 Malayan pangolins analyzed. Infected pangolins showed clinical signs and histological changes, and circulating antibodies against Pangolin-CoV reacted with the S protein of SARS-CoV-2. The isolation of a coronavirus that is highly related to SARS-CoV-2 in pangolins suggests that they have the potential to act as the intermediate host of SARS-CoV-2. The newly identified coronavirus in the most-trafficked mammal could represent a future threat to public health if wildlife trade is not effectively controlled.

Author notes: These authors contributed equally: Kangpeng Xiao, Junqiong Zhai

Keywords: SARS-CoV-2; COVID-19; Pangolins; Bats; Wildlife.

—–

#Evolutionary #Trajectory for the Emergence of Novel #Coronavirus #SARS-CoV-2 (Pathogens, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Pathogens, 9 (3) 2020 Mar 23

Evolutionary Trajectory for the Emergence of Novel Coronavirus SARS-CoV-2

Saif Ur Rehman 1, Laiba Shafique 1, Awais Ihsan 2 3, Qingyou Liu 1

Affiliations: 1 State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources of Guangxi University, Nanning 530005, China. 2 Department of Biosciences, COMSATS University Islamabad, Sahiwal Campus 57000, Pakistan. 3 College of Pharmacy, South Central University for Nationalities, Wuhan 430074, China.

PMID: 32210130 DOI: 10.3390/pathogens9030240

 

Abstract

Over the last two decades, the world experienced three outbreaks of coronaviruses with elevated morbidity rates. Currently, the global community is facing emerging virus SARS-CoV-2 belonging to Betacoronavirus, which appears to be more transmissible but less deadly than SARS-CoV. The current study aimed to track the evolutionary ancestors and different evolutionary strategies that were genetically adapted by SARS-CoV-2. Our whole-genome analysis revealed that SARS-CoV-2 was the descendant of Bat SARS/SARS-like CoVs and bats served as a natural reservoir. SARS-CoV-2 used mutations and recombination as crucial strategies in different genomic regions including the envelop, membrane, nucleocapsid, and spike glycoproteins to become a novel infectious agent. We confirmed that mutations in different genomic regions of SARS-CoV-2 have specific influence on virus reproductive adaptability, allowing for genotype adjustment and adaptations in rapidly changing environments. Moreover, for the first time we identified nine putative recombination patterns in SARS-CoV-2, which encompass spike glycoprotein, RdRp, helicase and ORF3a. Six recombination regions were spotted in the S gene and are undoubtedly important for evolutionary survival, meanwhile this permitted the virus to modify superficial antigenicity to find a way from immune reconnaissance in animals and adapt to a human host. With these combined natural selected strategies, SARS-CoV-2 emerged as a novel virus in human society.

Keywords: SARS-CoV; SARS-CoV-2; evolutionary strategies; genomic structure; mutations; phylogeny; recombination or reassortment.

Keywords: SARS-CoV-2; COVID-19; Evolution; Recombination.

——

Full- #genome #evolutionary #analysis of the novel #corona virus (2019-nCoV) rejects the hypothesis of emergence as a result of a recent #recombination event (Infect Genet Evol., abstract)

[Source: Infection, Genetics & Evolution, full page: (LINK). Abstract, edited.]

Infection, Genetics and Evolution / Volume 79, April 2020, 104212 / Short communication

Full-genome evolutionary analysis of the novel corona virus (2019-nCoV) rejects the hypothesis of emergence as a result of a recent recombination event

D. Paraskevis a, E.G. Kostaki a, G. Magiorkinis a, G. Panayiotakopoulos b, G. Sourvinos c, S. Tsiodras d

{a} Department of Hygiene Epidemiology and Medical Statistics, Medical School, National and Kapodistrian University of Athens, Athens, Greece; {b} National Public Health Organization (NPHO), Athens, Greece; {c} Laboratory of Clinical Virology, School of Medicine, University of Crete, Heraklion, Greece; {d} Medical School, National and Kapodistrian University of Athens, Athens, Greece

Received 27 January 2020, Accepted 28 January 2020, Available online 29 January 2020. DOI: https://doi.org/10.1016/j.meegid.2020.104212

 

Highlights

  • Full-genomic sequence analysis of the novel corona virus (2019-nCoV).
  • Phylogenetic and recombination analysis within the subgenus of sarbecovirus.
  • Evidence that the 2019-nCoV shows discordant clustering with the Bat_SARS-like coronavirus sequences.
  • Evidence that the hypothesis of emergence of 2019-nCoV as a result of a recent recombination event is rejected.

 

Abstract

Background

A novel coronavirus (2019-nCoV) associated with human to human transmission and severe human infection has been recently reported from the city of Wuhan in China. Our objectives were to characterize the genetic relationships of the 2019-nCoV and to search for putative recombination within the subgenus of sarbecovirus.

Methods

Putative recombination was investigated by RDP4 and Simplot v3.5.1 and discordant phylogenetic clustering in individual genomic fragments was confirmed by phylogenetic analysis using maximum likelihood and Bayesian methods.

Results

Our analysis suggests that the 2019-nCoV although closely related to BatCoV RaTG13 sequence throughout the genome (sequence similarity 96.3%), shows discordant clustering with the Bat_SARS-like coronavirus sequences. Specifically, in the 5′-part spanning the first 11,498 nucleotides and the last 3′-part spanning 24,341–30,696 positions, 2019-nCoV and RaTG13 formed a single cluster with Bat_SARS-like coronavirus sequences, whereas in the middle region spanning the 3′-end of ORF1a, the ORF1b and almost half of the spike regions, 2019-nCoV and RaTG13 grouped in a separate distant lineage within the sarbecovirus branch.

Conclusions

The levels of genetic similarity between the 2019-nCoV and RaTG13 suggest that the latter does not provide the exact variant that caused the outbreak in humans, but the hypothesis that 2019-nCoV has originated from bats is very likely. We show evidence that the novel coronavirus (2019-nCov) is not-mosaic consisting in almost half of its genome of a distinct lineage within the betacoronavirus. These genomic features and their potential association with virus characteristics and virulence in humans need further attention.

Keywords: Novel coronavirus  – Genomic sequence analysis – Phylogenetic analysis – Recombination – Origin – Molecular epidemiology

Keywords: 2019-nCoV; Genetics; Bats; Evolution; Betacoronavirus; Sarbecovirus.

——

Full-genome #Evolutionary #Analysis of the Novel #Coronavirus (2019-nCoV) Rejects the #Hypothesis of Emergence as a Result of a Recent #Recombination Event (Infect Genet Evol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Infect Genet Evol, 104212 2020 Jan 28 [Online ahead of print]

Full-genome Evolutionary Analysis of the Novel Corona Virus (2019-nCoV) Rejects the Hypothesis of Emergence as a Result of a Recent Recombination Event

D Paraskevis 1, E G Kostaki 2, G Magiorkinis 2, G Panayiotakopoulos 3, G Sourvinos 4, S Tsiodras 5

Affiliations: 1 Department of Hygiene Epidemiology and Medical Statistics, Medical School, National and Kapodistrian University of Athens, Athens, Greece. Electronic address: dparask@med.uoa.gr. 2 Department of Hygiene Epidemiology and Medical Statistics, Medical School, National and Kapodistrian University of Athens, Athens, Greece. 3 National Public Health Organization (NPHO), Athens, Greece. 4 Laboratory of Clinical Virology, School of Medicine, University of Crete, Heraklion, Greece. 5 Medical School, National and Kapodistrian University of Athens, Athens, Greece.

PMID: 32004758 DOI: 10.1016/j.meegid.2020.104212

 

Abstract

Background:

A novel coronavirus (2019-nCoV) associated with human to human transmission and severe human infection has been recently reported from the city of Wuhan in China. Our objectives were to characterize the genetic relationships of the 2019-nCoV and to search for putative recombination within the subgenus of sarbecovirus.

Methods:

Putative recombination was investigated by RDP4 and Simplot v3.5.1 and discordant phylogenetic clustering in individual genomic fragments was confirmed by phylogenetic analysis using maximum likelihood and Bayesian methods.

Results:

Our analysis suggests that the 2019-nCoV although closely related to BatCoV RaTG13 sequence throughout the genome (sequence similarity 96.3%), shows discordant clustering with the Bat_SARS-like coronavirus sequences. Specifically, in the 5′-part spanning the first 11,498 nucleotides and the last 3′-part spanning 24,341-30,696 positions, 2019-nCoV and RaTG13 formed a single cluster with Bat_SARS-like coronavirus sequences, whereas in the middle region spanning the 3′-end of ORF1a, the ORF1b and almost half of the spike regions, 2019-nCoV and RaTG13 grouped in a separate distant lineage within the sarbecovirus branch.

Conclusions:

The levels of genetic similarity between the 2019-nCoV and RaTG13 suggest that the latter does not provide the exact variant that caused the outbreak in humans, but the hypothesis that 2019-nCoV has originated from bats is very likely. We show evidence that the novel coronavirus (2019-nCov) is not-mosaic consisting in almost half of its genome of a distinct lineage within the betacoronavirus. These genomic features and their potential association with virus characteristics and virulence in humans need further attention.

Keywords: Analysis; Genomic sequence analysis; Molecular epidemiology; Novel coronavirus; Origin; Phylogenetic; Recombination.

Copyright © 2019. Published by Elsevier B.V.

Conflict of interest statement: Declaration of Competing Interest All authors report no conflict of interest related to the submitted work.

Keywords: 2019-nCoV; Bats; Recombination.

——-

#Enzootic #patterns of #MERS #coronavirus in imported #African and local #Arabian dromedary #camels: a prospective genomic study (Lancet Planet Health, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Lancet Planet Health. 2019 Dec 11. pii: S2542-5196(19)30243-8. doi: 10.1016/S2542-5196(19)30243-8. [Epub ahead of print]

Enzootic patterns of Middle East respiratory syndrome coronavirus in imported African and local Arabian dromedary camels: a prospective genomic study.

El-Kafrawy SA1, Corman VM2, Tolah AM3, Al Masaudi SB4, Hassan AM5, Müller MA6, Bleicker T7, Harakeh SM1, Alzahrani AA8, Alsaaidi GA8, Alagili AN9, Hashem AM10, Zumla A11, Drosten C12, Azhar EI13.

Author information: 1 Special Infectious Agent Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia. 2 Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Germany; Berlin Institute of Health, Institute of Virology, Berlin, Germany; German Centre for Infection Research, associated partner Charité, Berlin, Germany. 3 Special Infectious Agent Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; Department of Biological Science, Division of Microbiology, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia. 4 Department of Biological Science, Division of Microbiology, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia. 5 Special Infectious Agent Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia. 6 Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Germany; Berlin Institute of Health, Institute of Virology, Berlin, Germany; German Centre for Infection Research, associated partner Charité, Berlin, Germany; Martsinovsky Institute of Medical Parasitology, Tropical and Vector Borne Diseases, Sechenov University, Moscow, Russia. 7 Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Germany; Berlin Institute of Health, Institute of Virology, Berlin, Germany. 8 Directorate of Agriculture, Ministry of Environment Water and Agriculture, Makkah Region, Saudi Arabia. 9 Mammals Research Chair, Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia. 10 Special Infectious Agent Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia. 11 Department of Infection, Division of Infection and Immunity, Centre for Clinical Microbiology, University College London, London, UK; NIHR Biomedical Research Centre, University College London Hospitals, London, UK. 12 Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Germany; Berlin Institute of Health, Institute of Virology, Berlin, Germany; German Centre for Infection Research, associated partner Charité, Berlin, Germany. Electronic address: christian.drosten@charite.de. 13 Special Infectious Agent Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia; Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia. Electronic address: eazhar@kau.edu.sa.

 

Abstract

BACKGROUND:

The Middle East respiratory syndrome coronavirus (MERS-CoV) is a lethal zoonotic pathogen endemic to the Arabian Peninsula. Dromedary camels are a likely source of infection and the virus probably originated in Africa. We studied the genetic diversity, geographical structure, infection prevalence, and age-associated prevalence among camels at the largest entry port of camels from Africa into the Arabian Peninsula.

METHODS:

In this prospective genomic study, we took nasal samples from camels imported from Sudan and Djibouti into the Port of Jeddah in Jeddah, Saudi Arabia, over an almost 2-year period and local Arabian camels over 2 months in the year after surveillance of the port. We determined the prevalence of MERS-CoV infection, age-associated patterns of infection, and undertook phylogeographical and migration analyses to determine intercountry virus transmission after local lineage establishment. We compared all virological characteristics between the local and imported cohorts. We compared major gene deletions between African and Arabian strains of the virus. Reproductive numbers were inferred with Bayesian birth death skyline analyses.

FINDINGS:

Between Aug 10, 2016, and May 3, 2018, we collected samples from 1196 imported camels, of which 868 originated from Sudan and 328 from Djibouti, and between May 1, and June 25, 2018, we collected samples from 472 local camels, of which 189 were from Riyadh and 283 were from Jeddah, Saudi Arabia. Virus prevalence was higher in local camels than in imported camels (224 [47·5%] of 472 vs 157 [13·1%] of 1196; p<0·0001). Infection prevalence peaked among camels older than 1 year and aged up to 2 years in both groups, with 255 (66·9%) of 381 positive cases in this age group. Although the overall geographical distribution of the virus corresponded with the phylogenetic tree topology, some virus exchange was observed between countries corresponding with trade routes in the region. East and west African strains of the virus appear to be geographically separated, with an origin of west African strains in east Africa. African strains of the virus were not re-sampled in Saudi Arabia despite sampling approximately 1 year after importation from Africa. All local Arabian samples contained strains of the virus that belong to a novel recombinant clade (NRC) first detected in 2014 in Saudi Arabia. Reproduction number estimates informed by the sequences suggest sustained endemicity of NRC, with a mean Re of 1·16.

INTERPRETATION:

Despite frequent imports of MERS-CoV with camels from Africa, African lineages of MERS-CoV do not establish themselves in Saudi Arabia. Arabian strains of the virus should be tested for changes in virulence and transmissibility.

FUNDING:

German Ministry of Research and Education, EU Horizon 2020, and Emerging Diseases Clinical Trials Partnership.

Copyright © 2019 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license. Published by Elsevier Ltd.. All rights reserved.

PMID: 31843456 DOI: 10.1016/S2542-5196(19)30243-8

Keywords: MERS-CoV; Camels; Africa Region; Saudi Arabia; Recombination.

——

#Genetic #diversity of #MERS-CoV spike protein gene in #Saudi Arabia (J Infect Public Health, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

J Infect Public Health. 2019 Dec 9. pii: S1876-0341(19)30345-4. doi: 10.1016/j.jiph.2019.11.007. [Epub ahead of print]

Genetic diversity of MERS-CoV spike protein gene in Saudi Arabia.

Sohrab SS1, Azhar EI2.

Author information: 1 Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, 21589, Saudi Arabia. Electronic address: ssohrab@kau.edu.sa. 2 Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, 21589, Saudi Arabia; Medical Laboratory Technology Department, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, 21589, Saudi Arabia.

 

Abstract

BACKGROUND:

Middle East respiratory syndrome coronavirus (MERS-CoV) was primarily detected in 2012 and still causing disease in human and camel. Camel and bats have been identified as a potential source of virus for disease spread to human. Although, significant information related to MERS-CoV disease, spread, infection, epidemiology, clinical features have been published, A little information is available on the sequence diversity of Spike protein gene. The Spike protein gene plays a significant role in virus attachment to host cells. Recently, the information about recombinant MERS-CoV has been published. So, this work was designed to identify the emergence of any another recombinant virus in Jeddah, Saudi Arabia.

METHODS:

In this study samples were collected from both human and camels and the Spike protein gene was amplified and sequenced. The nucleotide and amino acid sequences of MERS-CoV Spike protein gene were used to analyze the recombination, genetic diversity and phylogenetic relationship with selected sequences from Saudi Arabia.

RESULTS:

The nucleotide sequence identity ranged from 65.7% to 99.8% among all the samples collected from human and camels from various locations in the Kingdom. The lowest similarity (65.7%) was observed in samples from Madinah and Dammam. The phylogenetic relationship formed different clusters with multiple isolates from various locations. The sample collected from human in Jeddah hospital formed a closed cluster with human samples collected from Buraydah, while camel sample formed a closed cluster with Hufuf isolates. The phylogenetic tree by using Aminoacid sequences formed closed cluster with Dammam, Makkah and Duba isolates. The amino acid sequences variations were observed in 28/35 samples and two unique amino acid sequences variations were observed in all samples analyzed while total 19 nucleotides sequences variations were observed in the Spike protein gene. The minor recombination events were identified in eight different sequences at various hotspots in both human and camel samples using recombination detection programme.

CONCLUSION:

The generated information from this study is very valuable and it will be used to design and develop therapeutic compounds and vaccine to control the MERS-CoV disease spread in not only in the Kingdom but also globally.

Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.

KEYWORDS: Camel; Genetic diversity; Human; MERS-CoV; Saudi Arabia

PMID: 31831395 DOI: 10.1016/j.jiph.2019.11.007

Keywords: MERS-CoV; Human; Camels; Recombination; Saudi Arabia.

——

#Serological evidence of #MERS-CoV and #HKU8-related #Coronavirus #coinfection in #Kenyan #camels (Emerg Microbes Infect., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Emerg Microbes Infect. 2019;8(1):1528-1534. doi: 10.1080/22221751.2019.1679610.

Serological evidence of MERS-CoV and HKU8-related CoV co-infection in Kenyan camels.

Zhang W1, Zheng XS1,2, Agwanda B3, Ommeh S4, Zhao K1,2, Lichoti J5, Wang N1, Chen J1,2, Li B1, Yang XL1, Mani S6, Ngeiywa KJ5,7, Zhu Y1, Hu B1, Onyuok SO1, Yan B1, Anderson DE6, Wang LF6, Zhou P1, Shi ZL1.

Author information: 1 CAS Key Laboratory of Special Pathogens, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences , Wuhan , People’s Republic of China. 2 University of Chinese Academy of Sciences , Beijing , People’s Republic of China. 3 Department of Zoology, National Museums of Kenya , Nairobi , Kenya. 4 Institute for Biotechnology Research, Jomo Kenyatta University of Agriculture and Technology , Nairobi , Kenya. 5 Directorate of Veterinary Services, State Department of Livestock, Ministry of Agriculture , Livestock and Fisheries , Kenya. 6 Programme in Emerging Infectious Diseases Duke-NUS Medical School , Singapore , Singapore. 7 Kenya Camel Association , Nairobi , Kenya.

 

Abstract

Dromedary camels are important reservoir hosts of various coronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV) that cause human infections. CoV genomes regularly undergo recombination during infection as observed in bat SARS-related CoVs. Here we report for the first time that only a small proportion of MERS-CoV receptor-binding domain positive (RBD) of spike protein positive camel sera in Kenya were also seropositive to MERS-CoV nucleocapsid (NP). In contrast, many of them contain antibodies against bat HKU8-related (HKU8r)-CoVs. Among 584 camel samples that were positive against MERS-CoV RBD, we found only 0.48 (8.22%) samples were also positive for NP. Furthermore, we found bat HKU8r-CoV NP antibody in 73 (12.5%) of the MERS-CoV RBD positive and NP negative samples, yet found only 3 (0.43%) of the HKU8r-CoV S1 antibody in the same samples. These findings may indicate co-infection with MERS-CoV and a HKU8r-CoV in camels. It may also raise the possibility of the circulation of a recombinant coronavirus virus with the spike of MERS-CoV and the NP of a HKU8r-CoV in Kenya. We failed to find molecular evidence of an HKU8r-CoV or a putative recombinant virus. Our findings should alert other investigators to look for molecular evidence of HKU8r-CoV or recombinants.

KEYWORDS: HKU8; MERS; bat; camel; coronavirus

PMID: 31645223 DOI: 10.1080/22221751.2019.1679610

Keywords: MERS-CoV; Coronavirus; Bats; Camels; Recombination; Kenya.

——

#Genetic and #phenotypic characterization of recently discovered #EV-D111 (PLoS Negl Trop Dis., abstract)

[Source: PLoS Neglected Tropical Diseases, full page: (LINK). Abstract, edited.]

OPEN ACCESS /  PEER-REVIEWED / RESEARCH ARTICLE

Genetic and phenotypic characterization of recently discovered enterovirus D type 111

Serge Alain Sadeuh-Mba , Marie-Line Joffret , Arthur Mazitchi, Marie-Claire Endegue-Zanga, Richard Njouom, Francis Delpeyroux, Ionela Gouandjika-Vasilache , Maël Bessaud

Published: October 17, 2019 / DOI: https://doi.org/10.1371/journal.pntd.0007797 / This is an uncorrected proof.

 

Abstract

Members of the species Enterovirus D (EV-D) remain poorly studied. The two first EV-D types (EV-D68 and EV-D70) have regularly caused outbreaks in humans since their discovery five decades ago but have been neglected until the recent occurrence of severe respiratory diseases due to EV-D68. The three other known EV-D types (EV-D94, EV-D111 and EV-D120) were discovered in the 2000s-2010s in Africa and have never been observed elsewhere. One strain of EV-D111 and all known EV-D120s were detected in stool samples of wild non-human primates, suggesting that these viruses could be zoonotic viruses. To date, EV-D111s are only known through partial genetic sequences of the few strains that have been identified so far. In an attempt to bring new pieces to the puzzle, we genetically characterized four EV-D111 strains (among the seven that have been reported until now). We observed that the EV-D111 strains from human samples and the unique simian EV-D111 strain were not phylogenetically distinct, thus suggesting a recent zoonotic transmission. We also discovered evidences of probable intertypic genetic recombination events between EV-D111s and EV-D94s. As recombination can only happen in co-infected cells, this suggests that EV-D94s and EV-D111s share common replication sites in the infected hosts. These sites could be located in the gut since the phenotypic analysis we performed showed that, contrary to EV-D68s and like EV-D94s, EV-D111s are resistant to acid pHs. We also found that EV-D111s induce strong cytopathic effects on L20B cells, a cell line routinely used to specifically detect polioviruses. An active circulation of EV-D111s among humans could then induce a high number of false-positive detection of polioviruses, which could be particularly problematic in Central Africa, where EV-D111 circulates and which is a key region for poliovirus eradication.

 

Author summary

Many examples of emergence of viruses that trigger severe diseases in humans are known. Emergence can be due to the sudden increase of the pathogenic power of a virus that had silently circulated into human populations for a long period; it can also be due to the cross-species transmission of a virus from its animal host to humans. The recent outbreaks of severe respiratory diseases due to enteroviruses D68 (EV-D68) brought to the light to potency of members of the species Enterovirus D (EV-D) to emerge as severe human pathogens.

By many aspects, EV-Ds are still mysterious: their natural history and epidemiology are poorly studied and even their main hosts remain unknown. For decades, EV-Ds were believed to infect mainly humans but recent data about EV-Ds identified in sub-Saharan Africa support their zoonotic origin. In an attempt to increase our knowledge about EV-Ds, we undertook genetic and phenotypic characterization of four EV-D111 isolates, a virus type that was recently uncovered in humans and in non-human primates in Central Africa. Our results show that EV-D111s are probably enteric viruses and evolve by exchanging genetic sequences with EV-D94.

___

Citation: Sadeuh-Mba SA, Joffret M-L, Mazitchi A, Endegue-Zanga M-C, Njouom R, Delpeyroux F, et al. (2019) Genetic and phenotypic characterization of recently discovered enterovirus D type 111. PLoS Negl Trop Dis 13(10): e0007797. https://doi.org/10.1371/journal.pntd.0007797

Editor: Abdallah M. Samy, Faculty of Science, Ain Shams University (ASU), EGYPT

Received: December 12, 2018; Accepted: September 18, 2019; Published: October 17, 2019

Copyright: © 2019 Sadeuh-Mba et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All sequence data are available at GenBank (accession numbers: MK032892-MK032898).

Funding: We are grateful for the financial support of the Institut Pasteur (PTR-276) and for the financial support of the World Health Organization. This work was supported by the European Virus Archive goes Global project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.

Keywords: Enterovirus; EV-D111; EV-D94; Recombination.

——

#Influenza Virus #Polymerase #Mutation Stabilizes a Foreign Gene Inserted into the Virus Genome by Enhancing the Transcription/Replication Efficiency of the Modified Segment (mBio, abstract)

[Source: mBio, full page: (LINK). Abstract, edited.]

Influenza Virus Polymerase Mutation Stabilizes a Foreign Gene Inserted into the Virus Genome by Enhancing the Transcription/Replication Efficiency of the Modified Segment

Yuri Furusawa, Shinya Yamada, Tiago Jose da Silva Lopes, Jayeeta Dutta, Zenab Khan, Divya Kriti, Harm van Bakel, Yoshihiro Kawaoka

J. S. Malik Peiris, Editor

DOI: 10.1128/mBio.01794-19

 

ABSTRACT

We previously attempted to establish a reporter influenza virus by inserting the gene for the Venus fluorescent protein into the NS segment of influenza A/Puerto Rico/8/34 (PR8, H1N1) virus to yield WT-Venus-PR8. Although the inserted Venus gene was deleted during serial passages of WT-Venus-PR8, we discovered that the PB2-E712D mutation stabilizes the Venus gene. Here, we explored the mechanisms by which Venus gene deletion occurs and how the polymerase mutation stabilizes the Venus gene. Deep sequencing analysis revealed that PB2-E712D does not cause an appreciable change in the mutation rate, suggesting that the stability of the Venus gene is not affected by polymerase fidelity. We found by using quantitative real-time PCR that WT-Venus-PR8 induces high-level interferon beta (IFN-β) expression. The induction of IFN-β expression seemed to result from the reduced transcription/replication efficiency of the modified NS segment in WT-Venus-PR8. In contrast, the transcription/replication efficiency of the modified NS segment was enhanced by the PB2-E712D mutation. Loss of the Venus gene in WT-Venus-PR8 appeared to be caused by internal deletions in the NS segment. Moreover, to further our understanding of the Venus stabilization mechanisms, we identified additional amino acid mutations in the virus polymerase complex that stabilize the Venus gene. We found that some of these amino acids are located near the template exit or the product exit of the viral polymerase, suggesting that these amino acids contribute to the stability of the Venus gene by affecting the binding affinity between the polymerase complex and the RNA template and product.

 

IMPORTANCE

The reverse genetics method of influenza virus generation has enabled us to generate recombinant viruses bearing modified viral proteins. Recombinant influenza viruses expressing foreign genes have become useful tools in basic research, and such viruses can be utilized as efficient virus vectors or multivalent vaccines. However, the insertion of a foreign gene into the influenza virus genome often impairs virus replication, and the inserted genes are unstable. Elucidation of the mechanisms of foreign gene stabilization will help us to establish useful recombinant influenza viruses.

Keywords: Influenza A; Recombination.

——

#Diversity of dromedary #camel #coronavirus #HKU23 in #African camels revealed multiple #recombination events among closely related #Betacoronaviruses of the subgenus #Embecovirus (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

Diversity of dromedary camel coronavirus HKU23 in African camels revealed multiple recombination events among closely related Betacoronaviruses of the subgenus Embecovirus.

Ray T. Y. So, Daniel K. W. Chu, Eve Miguel, Ranawaka A. P. M. Perera, Jamiu O. Oladipo, Ouafaa Fassi-Fihri, Gelagay Aylet, Ronald L. W. Ko, Ziqi Zhou, Mo-Sheung Cheng, Sulyman A. Kuranga, François L. Roger, Veronique Chevalier, Richard J. Webby, Patrick C. Y. Woo, Leo L. M. Poon, Malik Peiris

DOI: 10.1128/JVI.01236-19

 

ABSTRACT

Genetic recombination has frequently been observed in coronaviruses. Here, we sequenced multiple complete genomes of dromedary camel coronavirus HKU23 (DcCoV-HKU23) from Nigeria, Morocco and Ethiopia and identified several genomic positions indicative of cross species virus recombination events among other Betacoronaviruses of the subgenus Embecovirus (clade A β-CoVs). Recombinant fragments of a rabbit coronavirus (RbCoV-HKU14) were identified at the hemagglutinin esterase gene position. Homolog fragments of a rodent CoV were also observed at the 8.9 kDa open reading frame 4a at the 3′ end of the spike gene. The patterns of recombination varied geographically across the African region, highlighting a mosaic structure of DcCoV-HKU23 genomes circulating in dromedaries. Our results highlighted active recombination of coronaviruses circulating in dromedaries and is also relevant to the emergence and evolution of other Betacoronaviruses including MERS-coronavirus (MERS-CoV).

 

IMPORTANCE

Genetic recombination is often demonstrated in coronaviruses and can result in host range expansion or alteration in tissue tropism. Here, we showed interspecies recombination events of an endemic dromedary camel coronavirus HKU23 with other clade A Betacoronaviruses. Our results supported the possibility that the zoonotic pathogen, MERS-CoV, which also co-circulates in the same camel species, may have undergone similar recombination events facilitating its emergence or may do so in its future evolution.

Copyright © 2019 American Society for Microbiology. All Rights Reserved.

Keywords: Coronavirus; Betacoronavirus; Embecovirus; MERS-CoV; Camels; Africa Region; Recombination.

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