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. 2015 Dec 30;90(6):3253-6.
doi: 10.1128/JVI.02582-15.

Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus

Xing-Lou Yang  1 , Ben Hu  1 , Bo Wang  1 , Mei-Niang Wang  1 , Qian Zhang  1 , Wei Zhang  1 , Li-Jun Wu  1 , Xing-Yi Ge  1 , Yun-Zhi Zhang  2 , Peter Daszak  3 , Lin-Fa Wang  4 , Zheng-Li Shi  5
Affiliations

Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus

Xing-Lou Yang et al. J Virol. .

Abstract

We report the isolation and characterization of a novel bat coronavirus which is much closer to the severe acute respiratory syndrome coronavirus (SARS-CoV) in genomic sequence than others previously reported, particularly in its S gene. Cell entry and susceptibility studies indicated that this virus can use ACE2 as a receptor and infect animal and human cell lines. Our results provide further evidence of the bat origin of the SARS-CoV and highlight the likelihood of future bat coronavirus emergence in humans.

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Figures

FIG 1
FIG 1
Similarity plot based on the nucleotide sequence of the S gene of bat SL-CoV WIV16. S genes of human/civet SARS-CoVs and bat SL-CoV WIV1 were used as reference sequences, with a window of 200 bp and a step size of 20 bp under the Kimura model. CenterPos, center position.
FIG 2
FIG 2
Receptor analysis (A) and susceptibility test (B) results for bat SL-CoV WIV16. (A) HeLa cells with and without the expression of ACE2. ACE2 expression was detected with goat anti-human ACE2 antibody, followed by fluorescein isothiocyanate (FITC)-conjugated donkey anti-goat IgG. Virus replication was detected with rabbit antibody against the SL-CoV Rp3 nucleocapsid protein, followed by cyanine 3 (Cy3)-conjugated mouse anti-rabbit IgG. Nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole). The columns (from left to right) show staining of nuclei (blue), ACE2 expression (green), virus replication (red), and all three (merged triple-stain images). b, bat; c, civet; h, human. (B) Virus infection in A549, LLC-MK2, RSKT, PK15, H292, and Vero-E6 cells. The columns (from left to right) show staining of nuclei (blue), virus replication (red), and both nuclei and virus replication (merged double-stain images). A549 and H292, human lung cells; LLC-MK2, macaque kidney cells; RSKT, Chinese horseshoe bat kidney cells; PK15, pig kidney cells; VeroE6, African green monkey kidney cells.
FIG 3
FIG 3
One-step growth curve of bat SL-CoV WIV16 compared with that of WIV1. Vero E6 cells were infected with WIV16 or WIV1 at an MOI of 1. Supernatants were collected at 0, 12, 24, and 48 h postinfection. The viruses in the supernatant were determined by one-step reverse real-time PCR (n = 3). Virus RNA that had been extracted from a virus with a known titer was used to set up the standard curve. Error bars represent standard deviations. TCID50, 50% tissue culture infective doses.
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