China CDC Weekly

Notes from the Field

Isolation of 2019-nCoV from a Stool Specimen of a Laboratory-

Confirmed Case of the Coronavirus Disease 2019 (COVID-19)
Yong Zhang1,&;  Cao Chen1,&;  Shuangli Zhu1;  Chang Shu2;  Dongyan Wang1;  Jingdong Song1; 
Yang Song1;  Wei Zhen1;  Zijian Feng3;  Guizhen Wu1;  Jun Xu2,#;  Wenbo Xu1,#

The  novel  coronavirus  (2019-nCoV)  is  spreading
very  fast  in  Hubei  Province  of  China.  As  of  February
14,  2020,  51,986  confirmed  cases  (including
laboratory-confirmed  cases  and  clinically-confirmed
cases)  were  reported  in  Hubei  Province,  and  1,318  of
them  died.  Respiratory  droplets  and  contact
transmission  are  considered  to  be  the  most  important
routes of transmission of 2019-nCoV, but do not fully
account  for  the  occurrence  of  all  coronavirus  disease
2019  (COVID-19)  cases,  previously  known  as  novel
coronavirus pneumonia (NCP), and the reasons for the
rapid spread of this virus (1).
In  Biosafety  Level  3  (BSL-3)  Laboratory  of  the
National  Institute  for  Viral  Disease  Control  and
Prevention,  Vero  cells  were  used  for  viral  isolation
from  stool  samples  of  COVID-19  patients  sent  by
Heilongjiang  CDC.  A  2019-nCoV  strain  was  isolated
from  a  stool  specimen  of  a  laboratory-confirmed
COVID-19  severe  pneumonia  case,  who  experienced
onset  on  January  16,  2020  and  was  sampled  on
February 1, 2020. The interval between sampling and
onset  was  15  days.  The  full-length  genome  sequence
indicated  that  the  virus  had  high-nucleotide  similarity
(99.98%) to that of the first isolated novel coronavirus
isolated  from  Wuhan,  China  (Figure 1).  In  the  Vero
cells,  viral  particles  with  typical  morphology  of  a

LC522975.1-2019-nCoV/Japan/TY/WK-521/2020
69
LC522973.1-2019-nCoV/Japan/TY/WK-012/2020
71 LC522974.1-2019-nCoV/Japan/TY/WK-501/2020
MN997409.1-2019-nCoV/USA-AZ1/2020
MN975262.1-2019-nCoV HKU-SZ-005b/2020
MN938384.1-HKU-SZ-002a
MT019533.1-BetaCoV/Wuhan/IPBCAMS-WH-05/2020
MT019531.1-BetaCoV/Wuhan/IPBCAMS-WH-03/2019
MN994467.1-2019-nCoV/USA-CA1/2020
MN988669.1-2019-nCoV WHU02
MN996529.1-WIV05
MT019532.1-BetaCoV/Wuhan/IPBCAMS-WH-04/2019
LC522972.1-2019-nCoV/Japan/KY/V-029/2020
MT027064.1-2019-nCoV/USA-CA5/2020
MN996531.1-WIV07
MN908947.3-Wuhan-Hu-1
MT019529.1-BetaCoV/Wuhan/IPBCAMS-WH-01/2019
MN988713.1-2019-nCoV/USA-IL1/2020
79 MT027062.1-2019-nCoV/USA-CA3/2020
92 MT027063.1-2019-nCoV/USA-CA4/2020
2019-nCoV-HL002/HLJ/CHN/2020
NC 045512.2-Wuhan-Hu-1
MN996527.1-WIV02
MN996528.1-iWIV04
MN996530.1-WIV06
MT019530.1-BetaCoV/Wuhan/IPBCAMS-WH-02/2019
MT020781.1-nCoV-FIN-29-Jan-2020
MT039888.1-2019-nCoV/USA-MA1/2020
MT020881.1-2019-nCoV/USA-WA1-F6/2020
MT020880.1-2019-nCoV/USA-WA1-A12/2020
61 MN985325.1-2019-nCoV/USA-WA1/2020
MN988668.1-2019-nCoV WHU01
LC521925.1-2019-nCoV/Japan/AI/I-004/2020
MT039887.1-2019-nCoV/USA-WI1/2020
MT039873.1-HZ-1
MT007544.1-Australia/VIC01/2020
MN994468.1-2019-nCoV/USA-CA2/2020
MT039890.1-SNU01

0.000050

FIGURE 1. Phylogenetic tree based on the full-length genome sequences of 2019-nCoV. The sequence represented by the
red circle is the Heilongjiang sequence in this study.

Chinese Center for Disease Control and Prevention CCDC Weekly / Vol. 2 / No. 8 123
 China CDC Weekly
coronavirus  could  be  observed  under  the  electron
microscope (Figure 2).
100 nm
FIGURE 2. A  virus  particle  with  typical  morphology  of
coronavirus was observed using electron microscopy after
inoculating stool suspension into Vero cells.
These results confirm that COVID-19 patients have
live virus in stool specimens, which is a new finding in
the transmission routes of 2019-nCoV. In addition to
close contact and contact with respiratory secretions of
patients, the virus can also be transmitted through the
potential  fecal-oral  route.  This  means  that  stool
samples may contaminate hands, food, water, etc., and
may  cause  infection  by  invading  the  oral  cavity,
respiratory  mucosa,  conjunctiva,  etc.  This  virus  has
many  routes  of  transmission,  which  can  partially
explain  its  strong  transmission  and  fast  transmission
speed. This study also verified that the nucleic acids of
2019-nCoV can be detected from stool samples (2).
This  finding  has  important  public  health
124 CCDC Weekly / Vol. 2 / No. 8
significance.  Suggestions  to  strengthen  the  control  of
fecal  oral  transmission  of  2019-nCoV  include
strengthening  health  publicity  and  education;
maintaining  environmental  health  and  personal
hygiene;  drinking  boiled  water,  avoiding  raw  food
consumption, and implementing separate meal systems
in  epidemic  areas;  frequently  washing  hands  and
disinfecting of surfaces of objects in households, toilets,
public  places,  and  transportation  vehicles;  and
disinfecting the excreta and environment of patients in
medical  facilities  to  prevent  water  and  food
contamination from patients’ stool samples.
Acknowledgements
This  study  was  supported  by  the  National  Key
Technology  R&D  Programs  of  China  (Project  Nos.
2018ZX10713002 and 2017ZX10104001).
#
 Corresponding authors: Jun Xu, 1359865696@ qq.com; Wenbo Xu,
xuwb@ivdc.chinacdc.cn.
 
1
  National  Health  Commission  Key  Laboratory  for  Medical  Virology,
National  Institute  for  Viral  Disease  Control  and  Prevention,  Chinese
Center  for  Disease  Control  and  Prevention,  Beijing,  China;  2
Heilongjiang  Center  for  Disease  Control  and  Prevention,  Haerbin,
China;  3 Chinese Center for Disease Control and Prevention, Beijing,
China.
&
 Joint first authors.
Submitted: February 14, 2020; Accepted: February 14, 2020
References
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January  2020.  Euro  Surveill  2020;25(4).  http://dx.doi.org/10.2807/
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Chinese Center for Disease Control and Prevention