|
Objectives
1) to evaluate the specificity of 6 monoclonal
antibodies against the N protein of SARS CoV by studying the cross-reactivity
with human classical coronaviruses (HCoV-OC43, HCOV 229E), several
animal coronaviruses and other respiratory viruses,
2) to estimate age at first infection and age-specific
prevalence with classical human coronaviruses (HCoVOC43, HCOV 229E)
in the general Beijing population (the most severely affected city
with SARS patients in the world) and in France (not affected by
SARS CoV),
3) to study the potential cross-protection between
classical and SARS CoV using sera from patients with serologically
confirmed SARS disease.
Description of work
Specificity of 6 monoclonal antibodies against
the N protein of SARS CoV by studying the cross-eactivity with other
viral pathogens; indirect immunofluorescence assay (IFA) will be
performed by using anti-mouse immunoglobulins.
The tests will concern the following viral pathogens
:
• Human classical coronaviruses : HCoV-OC43, HCoV-229E
• Animal coronaviruses : BCV (Bovine CoV), MHV (Murine CoV),
TGEV (Transmissible
gastroenteritis porcine CoV)
• Others respiratory viruses : Influenza viruses A and B,
parainfluenza viruses 1, 2, 3, and 4, Respiratory syncytial viruses
A and B, human metapneumovirus, rhinoviruses (2 groups), enterovirus,
adenoviruses (A, B, C, D, E, F species), herpes viruses (HSV-1,
HSV-2, CMV, VZV), measles virus. Specimen collection for specificity
testing:
• prototype strains and wild type viral isolates of the selected
viruses (collected and stored at the Laboratory of Virology of Caen),
• fresh respiratory samples (nasal or nasopharyngeal aspirates,
pharyngeal swabs, broncho-alveolar lavage) from patients with acute
upper and lower respiratory tract infections (n = 200) collected
and stored at the Laboratory of Virology of Caen).
Study of cross-protection between classical coronaviruses
(HCoV-OC43 and HCoV-229E) and SARS-CoV:
Two types of HCoV (OC43 and 229E) antibodies will
be assessed :
• Total HCoV antibodies
- by immunofluorescence assay with prototype strains
• HCoV-OC43 isolated on HRT18 culture cells
• HCoV-229E isolated on MRC5 culture cells
- ELISA test with recombinant N protein expressed in baculovirus
system (development ongoing)
• Neutralizing antibodies by a microneutralization assay
Deliverables
D48 Specificity of monoclonal
antibodies against protein N of SARS CoV
D49 Seroprevalence study of contact
with classical human coronaviruses in Beijng and in France
D50 Potential cross-protection
between classical and SARS CoV
Milestones and expected result
M1-M6: protocol writing and collection
of sera for the seroprevalence study (from existing serabank)
M6-M24: development and validation
of tests
M18-M30: Synthesis of results
and manuscript preparation
|
