The Department of Emerging and Re-emerging Infections,

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The Department of Emerging and Re-emerging
Infections, Central Research Institute of
Epidemiology Moscow, Russia 2000-2004
Contacts platonov_at_pcr.ru
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West Nile fever Crimean-Congo hemorrhagic
fever Tick borne encephalitis Omsk hemorrhagic
fever Japanese encephalitis other flavivirus
infections hantaviral infection
are subjects we deal with
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Borreliosis Rickettsiosis Enteroviral
infection Meningococcal infection Haemophilus
influenzae type b Pneumococcal infection
are subjects we deal with
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treatment and control
epidemiology and surveillance
genodiagnostics and genotyping
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Russia Annual sum of effective temperatures (t
gt 10oC)
Moscow
200-500 500-1000
1000-1500 1500-2000 2000-2500
2500-3000 3000-3500
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West Nile fever classical and molecular
epidemiology, diagnosis, pathogenesis, treatment
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Molecular investigation of the simultaneous
outbreaks of West Nile fever in the USA and
Russia in 1999. (Bioterrorist's attack? No, the
attack of nature)
The beginning last week of July, The peak of
outbreak first week of September
?
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Genetic distance of West Nile virus
isolates based on complete nucleotide
sequence
"New York group" equine, flamingo, human, and
mosquito isolates in 1999, as well as the USA
strains from 2000-2002 and Israel strains from
1999-2001
Astrakhan strain, 1999
Italian strain, 1998 Kenyan strain, 1998
Mosquito isolate, Romania 1996 and human clinical
isolate, Volgograd 1999, (as well as the
Volgograd strains from humans, crows and
C.pipiens, 1999-2002)
Eg-101
Kunjin strains
Genetic distance
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The nearest relative of the Volgograd WN isolate
was the mosquito isolate from Romania-1996. The
strains differed from each other in 46 nucleotide
positions, 43 of them were located in coding
region. 35 mutations were silent so the strains
differed in 8 amino acid only. So the homology of
WN-Volgograd-4 and WN-Romania-1996M strains was
99.77 for aa sequence and 99.58 for nt
sequence.
Is the difference of 0.42 really small? Do these
strains belong to the same clone?
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Problem the origin of "Volgograd" variant of
West Nile virus. Is this variant endemic or are
several variants of West Nile virus introduced in
Volgograd region every year by migrating birds?
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West Nile fever in Russia in 1999-2003
WNF outbreak in 1999 Volgograd - 38 deaths
Astrakhan - 5 deaths Krasnodar - 3 deaths.
Astrakhan region
Volgograd
Krasnodar
Astrakhan
Number of WNF cases
95
49
?
33
24
12
?
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Genetic tree based on envelope gene fragment, 512
nt, of WN virus isolates from Volgograd
1999-2002
Volgograd WN clone appear to be endemic in
Volgograd region because the same isolates were
found in 1999-2002
patient-1999
C.pipiens pool-2001
two patients in 2000, one in 2002
patient-1999
patient-1999
9 of 1999
two crows in 2001
0.002 p-distance (one substitution)
crow-2001
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Selected publications and presentations
Platonov AE. West Nile encephalitis in Russia
1999-2001 Were we ready? Are we ready? In West
Nile virus Detection, surveillance, and control.
D.J.White and D.L.Morse, eds. New York, Annals of
the NYAS, 2001 V.951, pp.102-116. Platonov
A.E., Shipulin G.A., Shipulina O.Yu., et al.
Outbreak of West Nile infection - Volgograd
region, Russia, 1999. Emer. Infect. Dis. 2001 7
128-132 Platonov A.E., Karan L.S., Yazyshina
S.B., et al. Microheterogenicity of the Volgograd
clone of West Nile virus. International
Conference on Emerging Infectious Diseases.
Atlanta, March 24-27, 2002, p. 112. Platonov
A.E., Zhuravlev V.I., Lazorenko V.V. The weather
and the mosquito-borne infections What was wrong
in 1999 in Southern Russia? International
Conference on Emerging Infectious Diseases.
Atlanta, February 29 - March 3, 2004, p.63-64.
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Crimean-Congo hemorrhagic fever early pathogen
detection, viral load measurement, genotyping
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Crimean hemorrhagic fever (CHF) was diagnosed in
the Russian Federation since 1944. 334 human
cases were registered in the Rostov region from
1963 to 1971 years, 51 cases (15) were fatal.
CHF outbreaks have been reported also in the
Republics of Kalmykya and Dagestan, the
Stavropol, Astrakhan and Krasnodar regions
(colored in red) of the Southern Russia
Crimean
In 2000-2003 more than 200 CHF cases with 18
deaths were laboratory confirmed in Russia. Both
old endemic regions and new region (Volgograd -
colored in yellow) were affected. In the latter
region there were 18 CHF cases in 2000, 9 cases
in 2001, 4 cases in 2002-2003 the Crimean-Congo
hemorrhagic fever virus (CCHFV) RNA was found in
the ticks. The most affected region was
Stavropol region, where we have done our studies
on viral load at CCHF.
Congo
Volgograd
Stavropol
After the period of relative composure the CHF
epidemic situation has become worse last years.
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Crimean-Congo hemorrhagic fever
in Stavropol Region, population 2,655,000
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Laboratory diagnosis of Crimean-Congo hemorrhagic
fever
IgG diagnosis is possible
IgM diagnosis is possible
PCR diagnosis is possible
Critical time for treat- ment and prevention
-- diagnostic methods
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The decrease of viral load in patients
with Crimean-Congo hemorrhagic fever
Viral load, arbitrary units, logarithmic scale
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Viral load correlates with the severity of
Crimean-Congo hemorrhagic fever (day 3 day 5 of
hospitalization)
Viral load, arbitrary units, logarithmic scale
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Phylogenetic tree based on partial sequences of
the small S segment of CCHF isolates
South Europe Rostov, Volgograd,
Stavropol, Astrakhan, Kosovo, Albania
0.05 p-distance
Africa South Africa, Senegal, Nigeria, Mauritani
a, Burkina Faso, C.A.R. U.A.E.
South Africa, Uganda
Greece
U.A.E. - United Arab Emirates, Pakistan, Madagask
ar
Senegal, Maurinatia, Iran
China, Kazakhstan
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CCHF PCR-positive result in tick pools from
Volgograd
H.marginatum 962 ticks 155 pools 13
CHF-positive
H.scupense 220 ticks 22 pools 2
CHF-positive
D.marginatus 17 ticks 2 pools 0 CHF-positive
R.rossicus 26 ticks 2 pools 0 CHF-positive
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Selected publications and presentations.
Platonov A.E., Karan L.S., Yazyshina S.B., et
al. Molecular identification of Crimean-Congo
hemorrhagic fever virus in human clinical cases
in Southern Russia. International Conference on
Emerging Infectious Diseases. Atlanta, March
24-27, 2002, p. 83. Karan L.S., Sannikova I.V.,
Platonov A.E., et al. Viral load at Crimean-Congo
hemorrhagic fever and its clinical significance.
International Conference on Emerging Infectious
Diseases. Atlanta, February 29 - March 3, 2004,
p.125. Karan L.S., Shipulin G.A., Platonov A.E.
PCR-based laboratory diagnosis of Crimean-Congo
hemorrhagic fever. Clinical Laboratory
Diagnostics, 2003, No 10, p.50-54. PCR-based
assay for CCHF RNA detection is approved by the
State Institute for Biological Standards and
Control
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The advantages for physicians and
epidomiologists the diagnostics of unusual or
new pathogen
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Birds mortality was observed in 1997-2001 in
Southern Europe. The WNF infection was suspected
due to concurrent West Nile fever equine enzootic

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Genodiagnostics of infectious diseases (approach
2) non-specific probes
Possible infectious agents

direct sequencing of specific fragment CTTGGAGCAC
GGTATCTAGAGTTTGAAGCYYTGGGGTTC..
identification of pathogens both usual,
unusual, or new pathogens
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Phylogenetic tree of flaviviruses based on NS5
Denge fever
West Nile fever
Yellow fever
Japanese encephalitis
Tick-borne encephalitis
Negishi, LI
Omsk hemor- rhagic fever
Up to 100 flaviviruses may be identified using
NS5 gene
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Non-specific NS5-based assay for the detection of
genus Flavivirus RNA
Tick-borne encephalitis strains
West Nile fever strains
?-
?-
?
Omsk hemorrhagic fever strains
Japanese encephalitis strains
?-
?
?-
?
?
Also positive results with Langat, Powassan,
looping ill, Saint Louis encephalitis viruses
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Specific E gene-based assay for the detection of
Japanese encephalitis virus RNA
1 2 3 4 5 6 7 8 9 NC C- C
1 2 3 4 5 6 7 8 9 1011121314151617
C-C
Lines 1-17 avian brain samples C - positive
control
10 11 12 13 14 15 16 17 18 ?? ?- ?
Lines 1-5, 10-15 JE strains (positive) Line 6 -
SLE strains (negative) Line 7-9 - TBE strains
(negative) Line 16 - Powassan strain
(negative) Line 17 Langat strain
(negative) Line 18 Looping ill strain (negative)
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Genetic tree (E gene nucleotide sequences) of
avian JE isolates and reference strains of JEV,
WNFV, SLEV
Japanese encephalitis West Nile fever Saint
Louis encephalitis
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Birds mortality in 1997-2001 in Southern Europe
- Natural enzootic JE infection in Europe?
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West Nile Virus Shoot
Thank you