title: Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor.
abstract: The 2002-3 pandemic caused by severe acute respiratory syndrome coronavirus (SARS-CoV) was one of the most significant public health events in recent history. An ongoing outbreak of Middle East respiratory syndrome coronavirus suggests that this group of viruses remains a key threat and that their distribution is wider than previously recognized. Although bats have been suggested to be the natural reservoirs of both viruses, attempts to isolate the progenitor virus of SARS-CoV from bats have been unsuccessful. Diverse SARS-like coronaviruses (SL-CoVs) have now been reported from bats in China, Europe and Africa, but none is considered a direct progenitor of SARS-CoV because of their phylogenetic disparity from this virus and the inability of their spike proteins to use the SARS-CoV cellular receptor molecule, the human angiotensin converting enzyme II (ACE2). Here we report whole-genome sequences of two novel bat coronaviruses from Chinese horseshoe bats (family: Rhinolophidae) in Yunnan, China: RsSHC014 and Rs3367. These viruses are far more closely related to SARS-CoV than any previously identified bat coronaviruses, particularly in the receptor binding domain of the spike protein. Most importantly, we report the first recorded isolation of a live SL-CoV (bat SL-CoV-WIV1) from bat faecal samples in Vero E6 cells, which has typical coronavirus morphology, 99.9% sequence identity to Rs3367 and uses ACE2 from humans, civets and Chinese horseshoe bats for cell entry. Preliminary in vitro testing indicates that WIV1 also has a broad species tropism. Our results provide the strongest evidence to date that Chinese horseshoe bats are natural reservoirs of SARS-CoV, and that intermediate hosts may not be necessary for direct human infection by some bat SL-CoVs. They also highlight the importance of pathogen-discovery programs targeting high-risk wildlife groups in emerging disease hotspots as a strategy for pandemic preparedness.
authors.affiliation: 1] Center for Emerging Infectious Diseases, State Key Laboratory of Virology, Wuhan Institute of Virology of the Chinese Academy of Sciences, Wuhan 430071, China [2].
authors.forename: Xing-Yi
authors.initials: XY
authors.lastname: Ge
authors.forename: Jia-Lu
authors.initials: JL
authors.lastname: Li
authors.forename: Xing-Lou
authors.initials: XL
authors.lastname: Yang
authors.forename: Aleksei A
authors.initials: AA
authors.lastname: Chmura
authors.forename: Guangjian
authors.initials: G
authors.lastname: Zhu
authors.forename: Jonathan H
authors.initials: JH
authors.lastname: Epstein
authors.forename: Jonna K
authors.initials: JK
authors.lastname: Mazet
authors.forename: Ben
authors.initials: B
authors.lastname: Hu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Cheng
authors.initials: C
authors.lastname: Peng
authors.forename: Yu-Ji
authors.initials: YJ
authors.lastname: Zhang
authors.forename: Chu-Ming
authors.initials: CM
authors.lastname: Luo
authors.forename: Bing
authors.initials: B
authors.lastname: Tan
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhu
authors.forename: Gary
authors.initials: G
authors.lastname: Crameri
authors.forename: Shu-Yi
authors.initials: SY
authors.lastname: Zhang
authors.forename: Lin-Fa
authors.initials: LF
authors.lastname: Wang
authors.forename: Peter
authors.initials: P
authors.lastname: Daszak
authors.forename: Zheng-Li
authors.initials: ZL
authors.lastname: Shi
mesh_terms: D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D002681:China; D002685:Chiroptera / Q000821:virology*; D002522:Chlorocebus aethiops; D004197:Disease Reservoirs / Q000821:virology; D005243:Feces / Q000821:virology; D005455:Fluorescent Antibody Technique; D016679:Genome, Viral / Q000235:genetics; D058507:Host Specificity; D006801:Humans; D008969:Molecular Sequence Data; D058873:Pandemics / Q000517:prevention & control / Q000662:veterinary; D007703:Peptidyl-Dipeptidase A / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000648:ultrastructure; D045169:Severe Acute Respiratory Syndrome / Q000517:prevention & control / Q000635:transmission / Q000662:veterinary / Q000821:virology; D013045:Species Specificity; D064370:Spike Glycoprotein, Coronavirus / Q000737:chemistry / Q000378:metabolism; D014709:Vero Cells; D014771:Virion / Q000302:isolation & purification / Q000648:ultrastructure; D053586:Virus Internalization; D045949:Viverridae / Q000378:metabolism
chemical_list: D011991:Receptors, Virus; D064370:Spike Glycoprotein, Coronavirus; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; D000085962:Angiotensin-Converting Enzyme 2