???? CORONA VIRUSES

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???? CORONA VIRUSES

• ??
• ??????
• SARS-??????????
• ??????
• ????????????
• SARS?????????
• ?????????
• ????SARS?????SARS????
• ?????????????????
• ????

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• ?????? 33-35?
• ???????????
• 10-15?????????
• ?????(?????)
• ????????????
• HCoV-OC43, HCoV-229E
• ????RNA???? 80-160 nm
• ???????????
• ??? RNA 27,000-30,000 nt
• (???? 460?bp)
• (?? 30? bp)

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• SARS Severe Acute Respiratory Syndrome
• (??????????)
• Atypical pneumonia(?????)
• SARS coronavirus (WHO 3/16, 2003)
• Mortality rate 3-6(???)
• 43-55, older than 60 years
• African green monkey kidney (Vero E6) cells
• (??????????)
• SARS-HCoV, purifired, RNA genome was
  extracted(??????????RNA??)
• whole genome was sequenced(??????)

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????(??????)?????(?????) ?? ?? ?? ?? ????
?? ?? ??? ????? ????? ???????? ????????
X????????? X??????? ?????
????? ????????? ?????? ????? ?? ???????
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???RNA
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??RNA
???
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• ??????????
• SARS is a novel coronavirus
• Define a fourth class of coronavirus (group 4)
• Virus arose?
• harmless human coronavirus?
• animal?
• Development-
• PCR-based assay
• antiviral treatments
• neutralizing antibodies
• vaccine

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PCR-based assays ?PCR??DNA?????? ???????? (??????
???) (?????) ?????????primer
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HPC ??? HB ? B ?? DC ?? CH ?? CM ? CFB ? RD ??
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PCR- polymerase chain reaction ???????
?DNA????????? RT-PCR-?????????? reverse
transcription- polymerase chain reaction
?RNA????DNA,????DNA????
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Western blot Protein (antigen VS
antibody) Southern blot DNA hybridization Norther
n blot RNA hybridization
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• ????????
• Markets provide(?????)
• SCoV-like viruses amplify (??????)
• transmit to new hosts(??????)
• Further- surveillance on animals
• (???????????)

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????????? ??????????????????? ?????????SA
RS?????????? ?????????????????
???????,??????? ???
?????????,?????? ???????????????
?????,?????
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???(level 1, P1) ???????????????????? ???,?????-
12?????????1?4? (1)????????,???????? (2)?????????
? (3)???????????? (4)????????????? (5)????????????
? (6)????????????? (7)??????????
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???(level 2, P2) ?????????????????????, ????????
????? ??level 1???????? (1)???????????????? (2)??
?????????????????? (3)???????????????????? (4)????
????,??????????? ???????? (5)??1??2????????????
(6)????????????????????? (7)???????????? (8)?????
???? (9)??????????,?????? (10)???????????
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???(level 3, P3)  ???????????????,???????????
??level 1?level 2????,??? (1)???????,???????? (2
)?????????? (3)?????? (4)?????????????? (5)???????
??????? (6)??????????????????? (7)????????????????
???? ???,??????????? (8)???????????,??????????
(9)?????????
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???(level 4, P4)  ???????????????,???????????
??level 1?2?3????,??? (1)????????????????? (2)??
????????,??????? (3)??????????????? (4)???????????
???,?????????
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???? 03/08/03 ????????????? 03/14/03
???SARS 04/02/03 ??SARS????????,??P3??????
??????????? 04/03/03 ??SARS???????(hemop
hagocytosis) ?? 04/09/03 ??????,??????RT-PCR?????
????????? 04/11/03
????????(cytokine profile)????????
,??SARS????????(cytokine storm) 04/14/03
???????????(virus particle) 04/21/03
SARS??????????????????? 04/28/03
??SARS??????????????????? 05/04/03
?????SARS??????29714????????
,???312???????TW1
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• Treatment protocol (NTUH)????????
• Antibiotics ???
• Ribavirin (oral) ?????
• Corticosteroid ???
• Immunoglobulin ?????

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• SARS Diagnosis (??)
• Routine and Biochemistry ???????
• lymphopenia, elevated LDH..
• Lung pathology ?????
• Virus isolation ?????
• Vero E6 cells
• Nucleic acid detection ?????
• RT-PCR, Real-Time RT-PCR
• Serology ???(?????)
• Immunofluorescence, ELISA

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?? PCR ?????? ??????(??10?) ???? ???? ???? ?
???? ?????? ???PCR????? Real-Time PCR????????
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???????????lt?gt,??????????????,?????????lt?gt,???????
???,??????????? ???/????? (2003.04.28)
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Molecular epidemiology of SCoV in
Taiwan ?????? 11?
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? ? 1
? ? ? ?
? ?
? ? ?
? ? ?
? ? ? ? ?
? ? ? ? ?
? ? ?
? ? TW1
? ? ?
? ? 2
? ? ? ?
? ? ? ?
? ? ? ? ?
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The Big Question NowWill It Be Back?
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• SARS vaccine developmentEmerg Infect Dis. 2005
  Jul11(7) Jiang S, He Y, Liu S.
• Developing effective and safe vaccines is
  urgently needed to prevent infection by severe
  acute respiratory syndrome (SARS)-associated
  coronavirus (SARS-CoV). The inactivated SARS-CoV
  vaccine may be the first one available for
  clinical use because it is easy to generate
  however, safety is the main concern. The spike
  (S) protein of SARS-CoV is the major inducer of
  neutralizing antibodies, and the receptor-binding
  domain (RBD) in the S1 subunit of S protein
  contains multiple conformational neutralizing
  epitopes. This suggests that recombinant proteins
  containing RBD and vectors encoding the RBD
  sequence can be used to develop safe and
  effective SARS vaccines.

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The causative agent of severe acute respiratory
syndrome (SARS) has been identified as a new type
of coronavirus. Here, we have investigated the
ability of adenoviral delivery of codonoptimised
SARS-CoV strain Urbani structural antigens spike
protein S1 fragment, membrane protein, and
nucleocapsid protein to induce virus-specific
broad immunity in rhesus macaques. We immunised
rhesus macaques intramuscularly with a
combination of the three Ad5-SARS-CoV vectors or
a control vector and gave a booster vaccination
on day 28. The vaccinated animals all had
antibody responses against spike protein S1
fragment and T-cell responses against the
nucleocapsid protein. All vaccinated animals
showed strong neutralising antibody responses to
SARS-CoV infection in vitro. These results show
that an adenoviral-based vaccine can induce
strong SARS-CoV-specific immune responses in the
monkey, and hold promise for development of a
protective vaccine against the SARS causal
agent. Lancet 2003 362 189596
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• ??
• SARS????????????
• SARS???????????????
• ????????????????
• ??????????RT-PCR
• ???????????
• ?????????
• SARS??????