Preparedness for the detection of Preparedness for the detection of emerging-and re-emerging pathogens emerging-and re-emerging pathogens

in Croatiain CroatiaAlemka Markotić, MD, PhD

Associate professor

University Hospital for Infectious DiseasesDepartment for Research

Department for Cinical ImmunologyZagrebCroatia

HistoryHistory• The University Hospital for Infectious Diseases was founded in the spring was founded in the spring

of 1893of 1893, under the name of the City Hospital for Contagious DiseasesCity Hospital for Contagious Diseases

Central Diagnostic Laboratory

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HantavirusesHantaviruses

POZITIVAN

GRANIČAN

NEGATIVAN

Serology – acute case detection and Serology – acute case detection and sero-surveysero-survey

• Puumala and Dobrava viruses

• ELISA (IgM, IgG – reagents provided by CDC)

• POC

Markotić et al. Nature Med 1995Markotić et al. Nature Med 1996Markotić et al. J Med Virol 2002Markotić et al Scand Infect Dis 2002Miletić Medved M et al. Lancet 2002Markotić et al. Croat Med J 2002

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Markotić et al. J Med Virol 2002Cvetko et al. J Med Virol 2005Cvetko et al. Am J Trop Med Hyg 2006 Plyusnina A et al. J Med Virol 2011Tadin A et al. VBZD (under review)

Molecular detection – human and Molecular detection – human and rodents samplesrodents samples

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Puumala, Dobrava, Tula, Saarema viruses

Markotić A et al. J Gen Virol 2003

Markotić A et al. Coll Antropol 2007

Culture based Culture based techniquestechniques

VeroVero E6MRC-5

293-HEKTHP-1

Prim. monocytes

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Merdić E. et al., Infektološki glasnik (Croatian Journal of Infection) 2008; 1: 17-21.Out of 50 mosquito species recorded in Croatia, 15 have potential medical significance. Anopheles atroparvus, An. plumbeus, An. sacharovi, Culex pipiens, and Aedes albopictus are among the most significant species that are potential vectors in Croatia. Climatic changes play an important role in changing the condition of habitats, making it possible for new species to enter certain areas. The tiger mosquito was recorded in Croatia in 2004, and it can now be found in Istria, and more increasingly in Zadar, Šibenik, Split and Dubrovnik, with new sites found on the islands of Cres and Lošinj, which are very attractive tourist destinations.

• Combined Ag/Ab POC test

• OneStep RT-PCR

• Nested PCR

• Sequencing

• Virus isolation

1 2 3 4 1 2 3 4 tip DENVPozi

tivna

kon

trola

Paci

jent

ica

1

Paci

jent

ica

2

Mar

ker

AF311958/Brazil 1997

AF513110/Brazil 2001

AF311957/Brazil 1997

AF311956/Brazil 1997

AF514883/Paragvaj 2000

AF514885/Argentina 2000

AF226685/Brazil 1990

AF226687/Francuska Gujana 1989

AF514878/Paragvaj 2000

Pacijent 1

Pacijent 2

Pacijent 3

AY732474/Tajland 1980

EU179860/Bahrain 2006

AY762084/Singapur 1993

FJ225446/Sri Lanka 1997

FJ225448/Sri Lanka 2004

AF298807/Obala Bjelokosti 1999

DENV1 Genotip V

DENV1 Genotip II AF180817/Tajland 1964

AY858983/Indonezija 2004

U88535/Otok Nauru 1974

AB189121/Indonezija 1998

AB189120/Indonezija 1998

AB074761/Indonezija 1988

AB204803/Japan 2004

DQ285561/Sejseli 2004

AB195673/Sejseli 2003

DQ285560/Otok Reunion 2004

DENV1 Genotip IV

AB074760/Japan 1943

AF298808/Djibuti 1998

AF309641/Kambodza 1998

AF350498/Kina 1980

DENV1 Genotip I

DENV2 U87411/Tajland 1984

97

92

72

85

65

80

92

69

75

98

98

88

91

66

68

78

60

Dengue virus 1-4Dengue virus 1-4(human samples, mosquitos)(human samples, mosquitos)

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Tick-borne encephalitisTick-borne encephalitis(human samples, ticks)(human samples, ticks)

• ELISA• PCR• Sequencing• Virus isolation (in the pipeline)

• PCR• ELISA (temporary)

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West Nile VirusWest Nile Virus(human samples, mosquitos)(human samples, mosquitos)

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Chikungunya virusChikungunya virus(human samples, mosquitos)(human samples, mosquitos)

• PCR

• H1, H3, H1N1 2009• POC• PCR• Sequencing

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Influenza AInfluenza A(human samples)(human samples)

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In the pipeline – molecular dgIn the pipeline – molecular dg

• CCHF

• Other hemorrhagic fevers

• Poxvirus

• Rickettsia

• Virus isolation and immunopathogenesis research – ntl and intl research resources?

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Greetings from Croatia and thank you for the attention !

alemka.markotic@gmail.com