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Appropriate diagnosis of Zika virus infection: Italy North-West experience
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
particular to the area and this reveal it on the clinical features which increases its important in the studying. http://dx.doi.org/10.1016/j.jcv.2016.08.107 Abstract no: 142 Presentation at ESCV 2016: Poster 68 Appropriate diagnosis of Zika virus infection: Italy North-West experience E. Burdino 1,∗ , T. Allice 1 , M.G. Milia 1 , G. Gregori 1 , T. Ruggiero 1 , G. Calleri 2 , F. Lipani 3 , A. Lucchini 3 , G. Venturi 4 , V. Ghisetti 1 1
Laboratory of Microbiology and Virology, Amedeo di Savoia Hospital, Turin, Italy 2 Unit A of Infectious and Tropical Diseases, Amedeo di Savoia Hospital, Turin, Italy 3 Department of Infectious Diseases, Amedeo di Savoia Hospital, Turin, Italy 4 National Reference Laboratory for Arboviruses, Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy Background: After large outbreaks occurred in Micronesia, 2007, and in the Pacific Area, 2013, Zika virus (ZIKV) was reported in Brazil in early 2015 and subsequently in the Americas and Caribbean. Autochthonous cases of ZIKV infections have been worldwide reported from at least 45 countries and increasing number of imported cases has been observed in Europe and United States. The aim of the study was to evaluated ZIKV epidemiology in travelers recently returning from endemic areas to Piemonte, Italy North-West (4.2 million inhabitants) from January to May 2016. Methods: 68 samples (49 sera, 19 urine) were collected from 41 travelers returning from ZIKV endemic areas and referring to the regional Centre for Infectious Diseases, Amedeo di Savoia Hospital, Turin. Patients underwent laboratory examinations to rule out a tropical fever. Specific IgG and IgM antibodies to ZIKV were detected with ELISA IgM and IgG assay (Euroimmun, AG). Confirmatory Plaque Reduction Neutralization Tests (PRNTs) were performed at the Istituto Superiore di Sanità, Rome, Italy. Real Time Polymerase Chain Reaction (RT-PCR) for ZIKV was performed on serum/urine with two commercial assays: RealStar Zika virus RT-PC Kit (Altona Diagnostics) and Genesig Standard Kit (Primerdesign) validated with a ZIKV PCR standard (MR766 Zika virus strain) kindly provided by the Robert Koch Institute using 10-fold serial dilutions from 106 to 1 copy/l. Results: Recent ZIKV infection was identified in 3 out of 41 (7.3%) travelers. Patient 1 (male, 29 years old) reported arthralgia, retroorbital pain, severe itching and mild burning maculopapular rash 5 days after returning from Venezuela. Leucopenia was present. Serology for ZIKV was IgM positive and IgG negative. ZIKV RNA was not detected in blood (urine not available). A second serologic test performed 2 months later showed IgG seroconversion (169 RU/mL) and undetectable IgM. Patient 2 (female, 31 years old) 2 days after returning from Venezuela, reported chest and limbs papular rash with arthralgia but no fever. ZIKV serologic tests showed a low IgG titer (50 RU/mL) with undetectable IgM. ZIKV RNA was negative in blood (urine not available). A second serum sample was withdrawn 2 months later and showed increasing IgG titer to 250 RU/mL. PRNTs for Patient 1 and 2 were positive for ZIKV neutralizing antibodies (titer ≥ 1:10). Patient 3 (female, 40 years old) 3 days after returning from the Dominican Republic presented with chest and limbs pruriginous rash and fever lasting from 8 days before, with leucopenia. ZIKV RNA was detected in
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urine (5913 copies/ml); in serum a high IgG titer (106 RU/mL) with undetectable IgM was reported. Nine days later, ZIKV RNA was still positive in urine (310 copies/ml); IgG titer increased to 265 RU/mL. All patients tested negative for Dengue and Chikungunya viruses and completely recovered after few days. ZIKV RT-PCR detection limit was 46 copies/L for the Altona assay and 23 for Genesig [mean cycle threshold (Ct) values 37.3 and 37.5, respectively] according to the ZIKV MR766 standard. Conclusions: Returning travellers are sentinels of a rapidly changing epidemiology and require a prompt diagnosis and a careful surveillance for their implications in subsequent autochthonous transmission of the disease. In this contest, standardized molecular and serologic tests are mandatory for the appropriate diagnosis. http://dx.doi.org/10.1016/j.jcv.2016.08.108 Abstract no: 164 Presentation at ESCV 2016: Poster 69 Screening of emerging viral infections among risk groups Judit Deak 1,∗ , G. Kemenesi 2 , F. Jakab 2 1 Department of Clinical Microbiology, University of Szeged, Hungary 2 Virological Research Group, János Szentágothai Research Center, University of Pécs, Institute of Biology, Faculty of Sciences, University of Pécs, Pécs, Hungary
Background: Among viral infections, Hantaviruses and West Nile viruses (WNV) have been detected. Chikungunya virus, which can cause outbreaks in the temperate region, has been found in Italy. The members of Sandfly fever viruses (Genus: Phlebovirus): Sicilian, Naples, Toscana and Cyprus types have been detected in Italy, Portugal, Spain, France, Greece, Austria, Croatia and Turkey. Crimean-Congo hemorrhagic fever has been detected in the Balkan states, and dengue fever in Croatia, France and Norway. Aedes albopictus, the vector of yellow fever, is widespread among the European coastal regions and islands. The history of yellow fever and dengue fever in temperate regions confirms that the transmission of both diseases could recur. Patients and Methods: PCR and RT-PCR (TIB Molbiol, Berlin, Germany and Roche, Mannheim, Germany) methods have been introduced for the screening of the nucleic acids of Chikungunya, Crimean-Congo hemorrhagic fever, Dengue, Hanta, Sandfly fever, and West Nile viruses in healthy risk groups (hunters, fishers, gardeners and keepers in zoological garden) and blood donors in South Hungary. Indirect immunofluorescence (IIF) methods were performed with BIOCHIP slides (Euroimmun Med. Lab. AG., Lübeck, Germany). Results: PCR examinations proved negative both in risk groups and controls. Hantavirus Seul, Dobrava, and Puumala IgG antibodies proved positive in the cases of 5, 4 and 1 individuals, respectively. Sandfly fever viruses: Sicilian, Naples, Toscana and Cypres IgM were positive in 6, 2, 5 and 1,and IgG antibodies in 21, 17, 16 and 11 individuals. WNV IgM was positive in 3, and IgG in 22 cases. Chikungunya and Crimean-Congo IgM and IgG were negative. Dengue virus IgM was positive in 10 cases, while IgG was negative. Conclusions: The intensification of migration, the growth in the density of the population, the susceptibility to infectious diseases, the decline of human immunity in consequence of insolation (UV effect), and malnutrition all tend to make humanity sensitive to infectious illnesses. Prevention, recognition, early diagnosis and treatment are very important to counter local endemics and epidemics.
particular to the area and this reveal it on the clinical features which increases its important in the studying. http://dx.doi.org/10.1016/j.jcv.2016.08.107 Abstract no: 142 Presentation at ESCV 2016: Poster 68 Appropriate diagnosis of Zika virus infection: Italy North-West experience E. Burdino 1,∗ , T. Allice 1 , M.G. Milia 1 , G. Gregori 1 , T. Ruggiero 1 , G. Calleri 2 , F. Lipani 3 , A. Lucchini 3 , G. Venturi 4 , V. Ghisetti 1 1
Laboratory of Microbiology and Virology, Amedeo di Savoia Hospital, Turin, Italy 2 Unit A of Infectious and Tropical Diseases, Amedeo di Savoia Hospital, Turin, Italy 3 Department of Infectious Diseases, Amedeo di Savoia Hospital, Turin, Italy 4 National Reference Laboratory for Arboviruses, Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy Background: After large outbreaks occurred in Micronesia, 2007, and in the Pacific Area, 2013, Zika virus (ZIKV) was reported in Brazil in early 2015 and subsequently in the Americas and Caribbean. Autochthonous cases of ZIKV infections have been worldwide reported from at least 45 countries and increasing number of imported cases has been observed in Europe and United States. The aim of the study was to evaluated ZIKV epidemiology in travelers recently returning from endemic areas to Piemonte, Italy North-West (4.2 million inhabitants) from January to May 2016. Methods: 68 samples (49 sera, 19 urine) were collected from 41 travelers returning from ZIKV endemic areas and referring to the regional Centre for Infectious Diseases, Amedeo di Savoia Hospital, Turin. Patients underwent laboratory examinations to rule out a tropical fever. Specific IgG and IgM antibodies to ZIKV were detected with ELISA IgM and IgG assay (Euroimmun, AG). Confirmatory Plaque Reduction Neutralization Tests (PRNTs) were performed at the Istituto Superiore di Sanità, Rome, Italy. Real Time Polymerase Chain Reaction (RT-PCR) for ZIKV was performed on serum/urine with two commercial assays: RealStar Zika virus RT-PC Kit (Altona Diagnostics) and Genesig Standard Kit (Primerdesign) validated with a ZIKV PCR standard (MR766 Zika virus strain) kindly provided by the Robert Koch Institute using 10-fold serial dilutions from 106 to 1 copy/l. Results: Recent ZIKV infection was identified in 3 out of 41 (7.3%) travelers. Patient 1 (male, 29 years old) reported arthralgia, retroorbital pain, severe itching and mild burning maculopapular rash 5 days after returning from Venezuela. Leucopenia was present. Serology for ZIKV was IgM positive and IgG negative. ZIKV RNA was not detected in blood (urine not available). A second serologic test performed 2 months later showed IgG seroconversion (169 RU/mL) and undetectable IgM. Patient 2 (female, 31 years old) 2 days after returning from Venezuela, reported chest and limbs papular rash with arthralgia but no fever. ZIKV serologic tests showed a low IgG titer (50 RU/mL) with undetectable IgM. ZIKV RNA was negative in blood (urine not available). A second serum sample was withdrawn 2 months later and showed increasing IgG titer to 250 RU/mL. PRNTs for Patient 1 and 2 were positive for ZIKV neutralizing antibodies (titer ≥ 1:10). Patient 3 (female, 40 years old) 3 days after returning from the Dominican Republic presented with chest and limbs pruriginous rash and fever lasting from 8 days before, with leucopenia. ZIKV RNA was detected in
S55
urine (5913 copies/ml); in serum a high IgG titer (106 RU/mL) with undetectable IgM was reported. Nine days later, ZIKV RNA was still positive in urine (310 copies/ml); IgG titer increased to 265 RU/mL. All patients tested negative for Dengue and Chikungunya viruses and completely recovered after few days. ZIKV RT-PCR detection limit was 46 copies/L for the Altona assay and 23 for Genesig [mean cycle threshold (Ct) values 37.3 and 37.5, respectively] according to the ZIKV MR766 standard. Conclusions: Returning travellers are sentinels of a rapidly changing epidemiology and require a prompt diagnosis and a careful surveillance for their implications in subsequent autochthonous transmission of the disease. In this contest, standardized molecular and serologic tests are mandatory for the appropriate diagnosis. http://dx.doi.org/10.1016/j.jcv.2016.08.108 Abstract no: 164 Presentation at ESCV 2016: Poster 69 Screening of emerging viral infections among risk groups Judit Deak 1,∗ , G. Kemenesi 2 , F. Jakab 2 1 Department of Clinical Microbiology, University of Szeged, Hungary 2 Virological Research Group, János Szentágothai Research Center, University of Pécs, Institute of Biology, Faculty of Sciences, University of Pécs, Pécs, Hungary
Background: Among viral infections, Hantaviruses and West Nile viruses (WNV) have been detected. Chikungunya virus, which can cause outbreaks in the temperate region, has been found in Italy. The members of Sandfly fever viruses (Genus: Phlebovirus): Sicilian, Naples, Toscana and Cyprus types have been detected in Italy, Portugal, Spain, France, Greece, Austria, Croatia and Turkey. Crimean-Congo hemorrhagic fever has been detected in the Balkan states, and dengue fever in Croatia, France and Norway. Aedes albopictus, the vector of yellow fever, is widespread among the European coastal regions and islands. The history of yellow fever and dengue fever in temperate regions confirms that the transmission of both diseases could recur. Patients and Methods: PCR and RT-PCR (TIB Molbiol, Berlin, Germany and Roche, Mannheim, Germany) methods have been introduced for the screening of the nucleic acids of Chikungunya, Crimean-Congo hemorrhagic fever, Dengue, Hanta, Sandfly fever, and West Nile viruses in healthy risk groups (hunters, fishers, gardeners and keepers in zoological garden) and blood donors in South Hungary. Indirect immunofluorescence (IIF) methods were performed with BIOCHIP slides (Euroimmun Med. Lab. AG., Lübeck, Germany). Results: PCR examinations proved negative both in risk groups and controls. Hantavirus Seul, Dobrava, and Puumala IgG antibodies proved positive in the cases of 5, 4 and 1 individuals, respectively. Sandfly fever viruses: Sicilian, Naples, Toscana and Cypres IgM were positive in 6, 2, 5 and 1,and IgG antibodies in 21, 17, 16 and 11 individuals. WNV IgM was positive in 3, and IgG in 22 cases. Chikungunya and Crimean-Congo IgM and IgG were negative. Dengue virus IgM was positive in 10 cases, while IgG was negative. Conclusions: The intensification of migration, the growth in the density of the population, the susceptibility to infectious diseases, the decline of human immunity in consequence of insolation (UV effect), and malnutrition all tend to make humanity sensitive to infectious illnesses. Prevention, recognition, early diagnosis and treatment are very important to counter local endemics and epidemics.