Prsentation PowerPoint

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Crimean-Congo Haemorrhagic Fever DIAGNOSIS
Hervé Zeller National Reference Center - WHO
Collaborating Centre for Arboviruses and Viral
Haemorrhagic Fevers, Institut Pasteur, Lyon
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Viral Haemorrhagic Fevers
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Viral Haemorrhagic Fevers
Arenaviridae (Lassa, Junin, Machupo, Guanarito)
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Viral Haemorrhagic Fevers
Arenaviridae (Lassa, Junin, Machupo, Guanarito)
Filoviridae (Ebola, Marburg)
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Viral Haemorrhagic Fevers
Arenaviridae (Lassa, Junin, Machupo, Guanarito)
Envelopped RNA viruses
Bunyaviridae (CCHF, RVF, Hantaviruses)
Filoviridae (Ebola, Marburg)
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Family Genus VIRUS DISTRIBUTION Flaviviridae
Flavivirus Yellow Fever Africa South
America Dengue 1,2,3,4. Tropical
areas Omsk HF Russia Alkhurma Saudi
Arabia Kyasanur Forest HF India Bunyavirida
e Phlebovirus Rift Valley Fever Africa, Saudi
Arabia Nairovirus Crimean-Congo HF Africa,
Eurasia Hantavirus Hantan Dobrava Puumala
Eurasia Sin Nombre, Andes Americas
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Hemorrhages are inconstant Emerging part of the
iceberg Most frequently asymptomatic
infections
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Biosafety Issues Related to Haemorrhagic Fever
Viruses

• Criteria for classification
• Disease severity
• Transmissibility to laboratory workers
• Availability of treatment
• Availability of vaccine
• Classification BSL 1 to BSL4

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CCHF VIABILITY SENSITIVITY TO DESINFECTANTS
sodium hypochlorite 2, glutaraldehyde 2,
formaldehyde SENSITIVE TO DESSICATION INACTIVA
TION IRRADIATION UV TEMPERATURE 1 hour
60C not complete inactivation beta
propiolactone 4C not complete inactivation
Triton X100
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(No Transcript)
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CCHF Nairovirus genome   _________________________
____________________ Segment Nucleotides Amino
acids Protein __________________________________
___________ S 1659-1712 442-482
N M 4888 1551 G1 G2 NSm? L 12255 4036
L? _____________________________________________
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VHF SUSPECT CASE
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VHF SUSPECT CASE
Malaria Hepatitis Typhoidis
Toxicosis Septicemia Leptospirosis Rickettsiosis
 
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VHF SUSPECT CASE
Malaria Hepatitis Typhoidis
Toxicosis Septicemia Leptospirosis Rickettsiosis
 
Epidemiological data, risk evaluation biological
analysis, differential diagnostic
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VHF SUSPECT CASE
Malaria Hepatitis Typhoidis
Toxicosis Septicemia Leptospirosis Rickettsiosis
 
Epidemiological data, risk evaluation biological
analysis, differential diagnostic
Contact between clinicians and biologists
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CCHF laboratory data
LEUCOPENIA, particularly neutropenia THROMBOCYTOPE
NIA Hematocrite increases early then falls
down ASL, AST levels increases Proteinuria and
hematuria Mild azotemia, bilirubine increase
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CCHF laboratory diagnosis

• Viral detection blood specimen
• RT-PCR (nested)
• Cell culture (Vero E6 cells)

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CCHF laboratory diagnosis

• Viral detection (blood specimen)
• RT-PCR (nested)
• Cell culture (Vero E6 cells)
• Antibody detection (serum sample)
• IFA
• ELISA IgM (immuno-capture) IgG
• NT

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CCHF laboratory diagnosis
Viremia 10-12 days (although afebrile). Can be
detected by PCR up to day 16 By day 9 all
patients will have IgM or IgG antibody
Information needed DATE OF ONSET OF FEVER
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CCHF viral/antibody kinetics
IgM
IgG
viremia
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0
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RT-PCR
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Viral isolation
ELISA IgM IgG
IFA
IgM duration 2-3 months up to 6 months
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Primers for RT-PCR on S segment
From J. Smith, 1990
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RT-PCR /Southern blot hybridization/antibody
retrospective study
From Burt et al, J Virol Methods 1998, 70129-137
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RT-PCR /Southern blot hybridization/IFA antibody
26 samples from 19 patients from day 3-12 of
illness
From Burt et al, J Virol Methods 1998,
70129-137
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Hyalomma sp.
ticks
RT-PCR Viral isolation
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536 pb amplicons of the S fragment of CCHF genome
using primers CSDR3/CSDF2. RFLP with Hinf I, Hae
III, and Alu I endonucleases
Rapport IP Dakar 1993
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Turkey 2003
Phylogenetic analysis of 46 partial sequences
(219 bp) of the S segment of CCHF virus
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Phylogenetic analysis of 46 partial sequences
(219 bp) of the S segment of CCHF virus. Seven
major genetic groups. Strains from the Middle
and Far East and from different African regions
cluster in clearly separated groups. TURKEY
2003 Preliminary data 96-98 homology with
strains from the Balck Sea area and
Kosovo KOSOVO AF404507 STAVROPOL AF481802
DROSDOV U88412 ROSTOV AY277672
Drostein et al, J Clin Microbiol 2002, 40 1122
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National Reference Center - WHO Collaborating
Centre for Arboviruses and Viral Haemorrhagic
Fevers, Institut Pasteur, Lyon Marie-Claude
Georges Isabelle Schuffenecker Ingrid
Marendat Séverine Murri Hervé Zeller
BSL 3
BSL 4