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INSTITUTE OF PUBLIC HEALTH INSTITUTE OF PUBLIC HEALTH Institute for Public Health of Montenegro – Centre For Disease Control and Prevention

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Center for Medical Microbiology Center for Medical Microbiology Department for Department for Department for Department for Molecular Diagnostics Molecular Diagnostics Virology and Serology Virology and Serology Molecular diagnostic of Serodiagnostic of most important medical most important medical microorganisms microorganisms pathogens Preparing for cell culture and virus isolation

• Assoc. Prof. Assoc. Prof. Boban Boban Mugo Mugoš ša, a, responsible responsible person person person person • Assoc. Prof. Assoc. Prof. Zoran Zoran Vratnica Vratnica, key researcher , key researcher • Dr. Dr. Danijela Danijela Vujo Vujoš ševi ević ć, researcher , researcher • Dr. Dr. Sanja Sanja Medenica Medenica, researcher , researcher • Dr. Dr. Neboj Nebojš ša a Sekuli Sekulić ć, researcher , researcher

University of Montenegro Biotechnical faculty WP1c Detection of pathogens in mosquitoes and humans

WP1c - Realized activities VISITS OF IPH RESEARCHERS TO INSTITUTE "ISTITUTO SPERIMENTALE ZOOPROFILATTICO REGGIO EMILIA" (IZSLER), 14 JULY - 01 AUGUST 2014

Preparation of mosquitoes pools for RNA extraction; Random primer Retro-Transcription reaction protocol assessment; Real Time PCR protocols and reactions set-up; Real Time PCR protocols and reactions set-up; Real Time PCR protocols and results interpretation; RNA extraction with semi-automatic RNA/DNA extractor.

RNA extraction Semi-automatic extractor Manualy (TRIzol LS Reagent)

Random primer Real Time PCR reactions set-up and results interpretation Retro-Transcription reaction

Results of molecular analyses from Montenegrian mosquites

Pathogen IgM IgG Microscopy Real-Time PCR ELISA ELISA WNV 48 tests 48 tests / / (all negative) (all negative) Borrelia burgdorferi Borrelia burgdorferi 838 tests 838 tests 840 tests 840 tests / / 76 tests 76 tests (79 positive) (59 positive) (all negative) Leishmania spp. / / 22 / (all negative) Leishmania donovani / 143 tests / / (10 positive) Plasmodium spp / / 14 / (all negative)

INSTITUTE OF PUBLIC HEALTH MONTENEGRO INSTITUTE OF PUBLIC HEALTH MONTENEGRO Centre for Disease Control and Prevention Mandatory reporting diseases - electronic database 2014 Lyme boreliosis – six reports of disease Leishmaniasis – three reports of disease Infectio West Nile – no registered cases Malaria – no registered cases

Equipment for IPH ELISA Analyzer I-2P QuantStudio™ 7 Flex Real-Time EUROIMMUN PCR System, Applied Biosystems

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