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 history1. An ongoing outbreak of Middle East respiratory syndrome coronavirus (MERS-CoV)2 suggests that this group of viruses remains a major threat and that their distribution is wider than previously recognized. Although bats have been suggested as the natural reservoirs of both viruses3–5, 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 Africa5–8, but none are considered a direct progenitor of SARS-CoV because of their phylogenetic disparity from this virus and the inability of their spike proteins (S) to use the SARS-CoV cellular receptor molecule, the human angiotensin converting enzyme II (ACE2)9,10. Here, we report whole genome sequences of two novel bat CoVs 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 CoVs, particularly in the receptor binding domain (RDB) of the S protein. Most importantly, we report the first recorded isolation of a live SL-CoV (bat SL-CoV-WIV1) from bat fecal samples in Vero E6 cells, which has typical coronavirus morphology, 99.9% sequence identity to Rs3367 and uses the ACE2s from human, civet and Chinese horseshoe bat 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.

A meta-analysis to evaluate the effectiveness of real-time PCR for diagnosing novel coronavirus infections

“…Of these, one patient was hospitalized in an intensive care unit and another patient died; they both appeared to have severe acute respiratory infection combined with renal failure (Assiri et al, 2013;Scobey et al, 2013;Martinez et al, 2014). World health organization (WHO) determined that the pathogen is a novel coronavirus with many similarities to SARS virus (Ge et al, 2013). A British scientist released its complete sequence recently (Corman et al, 2012).…”

Bats: carriers of zoonotic viral and emerging infectious diseases

“…In mouse, chimeric virus developed severe pathogenesis which was found untreatable with anti-SARS immunotherapeutics. Moreover, chimeric virus replicated in primary human airway cell line and produced an equivalent titer of SARS-CoV outbreak from human (Ge et al, 2013;Menachery et al, 2015) which indicates a vital threat of reemergence of human SARS-CoV from wild bat population.…”

Middle East Respiratory Syndrome

“…SARS-CoV most likely originated in bats 21,2223 and adapted to non-bat ACE2 variants as it crossed species to infect humans 24,25 . Most notably, there were changes in the surface (S) glycoproteinthat enhanced binding to the human ACE2 receptor.…”