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Dr. Serena Marchi
University of Siena

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Communication
Published: 12 July 2021 in Viruses
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The recent spreading of new SARS-CoV-2 variants, carrying several mutations in the spike protein, could impact immune protection elicited by natural infection or conferred by vaccination. In this study, we evaluated the neutralizing activity against the viral variants that emerged in the United Kingdom (B.1.1.7), Brazil (P.1), and South Africa (B.1.351) in human serum samples from hospitalized patients infected by SARS-CoV-2 during the first pandemic wave in Italy in 2020. Of the patients studied, 59.5% showed a decrease (≥2 fold) in neutralizing antibody titer against B.1.1.7, 83.3% against P.1, and 90.5% against B.1.351 with respect to the original strain. The reduction in antibody titers against all analyzed variants, and in particular P.1 and B.1.351, suggests that previous symptomatic infection might be not fully protective against exposure to SARS-CoV-2 variants carrying a set of relevant spike mutations.

ACS Style

Claudia Trombetta; Serena Marchi; Simonetta Viviani; Alessandro Manenti; Linda Benincasa; Antonella Ruello; Emilio Bombardieri; Ilaria Vicenti; Maurizio Zazzi; Emanuele Montomoli. Serum Neutralizing Activity against B.1.1.7, B.1.351, and P.1 SARS-CoV-2 Variants of Concern in Hospitalized COVID-19 Patients. Viruses 2021, 13, 1347 .

AMA Style

Claudia Trombetta, Serena Marchi, Simonetta Viviani, Alessandro Manenti, Linda Benincasa, Antonella Ruello, Emilio Bombardieri, Ilaria Vicenti, Maurizio Zazzi, Emanuele Montomoli. Serum Neutralizing Activity against B.1.1.7, B.1.351, and P.1 SARS-CoV-2 Variants of Concern in Hospitalized COVID-19 Patients. Viruses. 2021; 13 (7):1347.

Chicago/Turabian Style

Claudia Trombetta; Serena Marchi; Simonetta Viviani; Alessandro Manenti; Linda Benincasa; Antonella Ruello; Emilio Bombardieri; Ilaria Vicenti; Maurizio Zazzi; Emanuele Montomoli. 2021. "Serum Neutralizing Activity against B.1.1.7, B.1.351, and P.1 SARS-CoV-2 Variants of Concern in Hospitalized COVID-19 Patients." Viruses 13, no. 7: 1347.

Journal article
Published: 05 July 2021 in Pathogens
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Although in humans West Nile virus is mainly the cause of mild or sub-clinical infections, in some cases a neuroinvasive disease may occur predominantly in the elderly. In Italy, several cases of West Nile virus infection are reported every year. Tuscany was the first Italian region where the virus was identified; however, to date only two cases of infection have been reported in humans. This study aimed at evaluating the prevalence of antibodies against West Nile virus in the area of Siena Province to estimate the recent circulation of the virus. Human serum samples collected in Siena between 2016 and 2019 were tested for the presence of antibodies against West Nile virus by ELISA. ELISA positive samples were further evaluated using immunofluorescence, micro neutralization, and plaque reduction neutralization assays. In total, 1.9% (95% CI 1.2–3.1) and 1.4% (95% CI 0.8–2.4) of samples collected in 2016–2017 were positive by ELISA and immunofluorescence assay, respectively. Neutralizing antibodies were found in 0.7% (95% CI 0.3–1.5) of samples. Additionally, 0.9% (95% CI 0.4–1.7) and 0.65% (95% CI 0.3–1.45) of samples collected in 2018–2019 were positive by ELISA and immunofluorescence assay, respectively. The prevalence of neutralizing antibodies was 0.5% (95% CI 0.2–1.3). Although no human cases of West Nile infection were reported in the area between 2016 and 2019 and virus prevalence in the area of Siena Province was as low as less than 1%, the active asymptomatic circulation confirms the potential concern of this emergent virus for human health.

ACS Style

Serena Marchi; Emanuele Montomoli; Simonetta Viviani; Simone Giannecchini; Maria Stincarelli; Gianvito Lanave; Michele Camero; Caterina Alessio; Rosa Coluccio; Claudia Trombetta. West Nile Virus Seroprevalence in the Italian Tuscany Region from 2016 to 2019. Pathogens 2021, 10, 844 .

AMA Style

Serena Marchi, Emanuele Montomoli, Simonetta Viviani, Simone Giannecchini, Maria Stincarelli, Gianvito Lanave, Michele Camero, Caterina Alessio, Rosa Coluccio, Claudia Trombetta. West Nile Virus Seroprevalence in the Italian Tuscany Region from 2016 to 2019. Pathogens. 2021; 10 (7):844.

Chicago/Turabian Style

Serena Marchi; Emanuele Montomoli; Simonetta Viviani; Simone Giannecchini; Maria Stincarelli; Gianvito Lanave; Michele Camero; Caterina Alessio; Rosa Coluccio; Claudia Trombetta. 2021. "West Nile Virus Seroprevalence in the Italian Tuscany Region from 2016 to 2019." Pathogens 10, no. 7: 844.

Journal article
Published: 05 July 2021 in European Journal of Medicinal Chemistry
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The worldwide circulation of different viruses coupled with the increased frequency and diversity of new outbreaks, strongly highlight the need for new antiviral drugs to quickly react against potential pandemic pathogens. Broad-spectrum antiviral agents (BSAAs) represent the ideal option for a prompt response against multiple viruses, new and re-emerging. Starting from previously identified anti-flavivirus hits, we report herein the identification of promising BSAAs by submitting the multi-target 2,6-diaminopurine chemotype to a system-oriented optimization based on phenotypic screening on cell cultures infected with different viruses. Among the synthesized compounds, 6i showed low micromolar potency against Dengue, Zika, West Nile and Influenza A viruses (IC50 = 0.5–5.3 μM) with high selectivity index. Interestingly, 6i also inhibited SARS-CoV-2 replication in different cell lines, with higher potency on Calu-3 cells that better mimic the SARS-CoV-2 infection in vivo (IC50 = 0.5 μM, SI = 240). The multi-target effect of 6i on flavivirus replication was also analyzed in whole cell studies (in vitro selection and immunofluorescence) and against isolated host/viral targets.

ACS Style

Ilaria Vicenti; Maria Grazia Martina; Adele Boccuto; Marta De Angelis; Giorgia Giavarini; Filippo Dragoni; Serena Marchi; Claudia Maria Trombetta; Emmanuele Crespan; Giovanni Maga; Cecilia Eydoux; Etienne Decroly; Emanuele Montomoli; Lucia Nencioni; Maurizio Zazzi; Marco Radi. System-oriented optimization of multi-target 2,6-diaminopurine derivatives: Easily accessible broad-spectrum antivirals active against flaviviruses, influenza virus and SARS-CoV-2. European Journal of Medicinal Chemistry 2021, 224, 113683 .

AMA Style

Ilaria Vicenti, Maria Grazia Martina, Adele Boccuto, Marta De Angelis, Giorgia Giavarini, Filippo Dragoni, Serena Marchi, Claudia Maria Trombetta, Emmanuele Crespan, Giovanni Maga, Cecilia Eydoux, Etienne Decroly, Emanuele Montomoli, Lucia Nencioni, Maurizio Zazzi, Marco Radi. System-oriented optimization of multi-target 2,6-diaminopurine derivatives: Easily accessible broad-spectrum antivirals active against flaviviruses, influenza virus and SARS-CoV-2. European Journal of Medicinal Chemistry. 2021; 224 ():113683.

Chicago/Turabian Style

Ilaria Vicenti; Maria Grazia Martina; Adele Boccuto; Marta De Angelis; Giorgia Giavarini; Filippo Dragoni; Serena Marchi; Claudia Maria Trombetta; Emmanuele Crespan; Giovanni Maga; Cecilia Eydoux; Etienne Decroly; Emanuele Montomoli; Lucia Nencioni; Maurizio Zazzi; Marco Radi. 2021. "System-oriented optimization of multi-target 2,6-diaminopurine derivatives: Easily accessible broad-spectrum antivirals active against flaviviruses, influenza virus and SARS-CoV-2." European Journal of Medicinal Chemistry 224, no. : 113683.

Journal article
Published: 07 April 2021 in International Journal of Environmental Research and Public Health
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In Italy, the influenza season lasts from October until April of the following year. Influenza A and B viruses are the two viral types that cocirculate during seasonal epidemics and are the main causes of respiratory infections. We analyzed influenza A and B viruses in samples from hospitalized patients at Le Scotte University Hospital in Siena (Central Italy). From 2015 to 2020, 182 patients with Severe Acute Respiratory Infections were enrolled. Oropharyngeal swabs were collected from patients and tested by means of reverse transcriptase-polymerase chain reaction to identify influenza A(H3N2), A(H1N1)pdm09 and B. Epidemiological and virological surveillance remain an essential tool for monitoring circulating viruses and possible mismatches with seasonal vaccine strains, and provide information that can be used to improve the composition of influenza vaccines.

ACS Style

Ilaria Manini; Andrea Camarri; Serena Marchi; Claudia Trombetta; Ilaria Vicenti; Filippo Dragoni; Giacomo Lazzeri; Giovanni Bova; Emanuele Montomoli; Pier Capecchi. Surveillance for Severe Acute Respiratory Infections among Hospitalized Subjects from 2015/2016 to 2019/2020 Seasons in Tuscany, Italy. International Journal of Environmental Research and Public Health 2021, 18, 3875 .

AMA Style

Ilaria Manini, Andrea Camarri, Serena Marchi, Claudia Trombetta, Ilaria Vicenti, Filippo Dragoni, Giacomo Lazzeri, Giovanni Bova, Emanuele Montomoli, Pier Capecchi. Surveillance for Severe Acute Respiratory Infections among Hospitalized Subjects from 2015/2016 to 2019/2020 Seasons in Tuscany, Italy. International Journal of Environmental Research and Public Health. 2021; 18 (8):3875.

Chicago/Turabian Style

Ilaria Manini; Andrea Camarri; Serena Marchi; Claudia Trombetta; Ilaria Vicenti; Filippo Dragoni; Giacomo Lazzeri; Giovanni Bova; Emanuele Montomoli; Pier Capecchi. 2021. "Surveillance for Severe Acute Respiratory Infections among Hospitalized Subjects from 2015/2016 to 2019/2020 Seasons in Tuscany, Italy." International Journal of Environmental Research and Public Health 18, no. 8: 3875.

Communication
Published: 17 February 2021 in Vaccines
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Equine influenza (EI) is a highly contagious acute respiratory disease of equines that is caused mainly by the H3N8 subtype of influenza A virus. Vaccinating horses against EI is the most effective strategy to prevent the infection. The current study aimed to compare the kinetics of EI-specific humoral- and cell-mediated immunity (CMI) in horses receiving either identical or mixed vaccinations. Two groups of horses were previously (six months prior) vaccinated with either Calvenza 03 EIV EHV® (G1) or Fluvac Innovator® (G2) vaccine. Subsequently, both groups received a booster single dose of Calvenza 03 EIV EHV®. Immune responses were assessed after 10 weeks using single radial hemolysis (SRH), virus neutralization (VN), and EliSpot assays. Our results revealed that Calvenza-03 EIV/EHV®-immunized horses had significantly higher protective EI-specific SRH antibodies and VN antibodies. Booster immunization with Calvenza-03 EIV/EHV® vaccine significantly stimulated cell-mediated immune response as evidenced by significant increase in interferon-γ-secreting peripheral blood mononuclear cells. In conclusion, Calvenza-03 EIV/EHV® vaccine can be safely and effectively used for booster immunization to elicit optimal long persisting humoral and CMI responses even if the horses were previously immunized with a heterogeneous vaccine.

ACS Style

Selvaraj Pavulraj; Tobias Bergmann; Claudia Trombetta; Serena Marchi; Emanuele Montomoli; Sidi Alami; Roberto Ragni-Alunni; Nikolaus Osterrieder; Walid Azab. Immunogenicity of Calvenza-03 EIV/EHV® Vaccine in Horses: Comparative In Vivo Study. Vaccines 2021, 9, 166 .

AMA Style

Selvaraj Pavulraj, Tobias Bergmann, Claudia Trombetta, Serena Marchi, Emanuele Montomoli, Sidi Alami, Roberto Ragni-Alunni, Nikolaus Osterrieder, Walid Azab. Immunogenicity of Calvenza-03 EIV/EHV® Vaccine in Horses: Comparative In Vivo Study. Vaccines. 2021; 9 (2):166.

Chicago/Turabian Style

Selvaraj Pavulraj; Tobias Bergmann; Claudia Trombetta; Serena Marchi; Emanuele Montomoli; Sidi Alami; Roberto Ragni-Alunni; Nikolaus Osterrieder; Walid Azab. 2021. "Immunogenicity of Calvenza-03 EIV/EHV® Vaccine in Horses: Comparative In Vivo Study." Vaccines 9, no. 2: 166.

Journal article
Published: 05 November 2020 in Vaccines
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After the influenza H1N1 pandemic of 2009, the seasonal A/Brisbane/59/2007 strain was replaced by the A/California/07/2009 strain for the influenza virus vaccine composition. After several seasons with no indications on the occurrence of antigenic drift, A/Michigan/45/2015 was chosen as the H1N1 vaccine strain for the 2017/2018 season. Since the immune response to influenza is shaped by the history of exposure to antigenically similar strains, the potential cross-protection between seasonal human influenza vaccine strains and the emerging pandemic strains was investigated. Human serum samples were tested by hemagglutination inhibition and single radial hemolysis assays against A/Brisbane/59/2007, A/California/07/2009, and A/Michigan/45/2015 strains. Strong cross-reactions between A/California/07/2009 and A/Michigan/45/2015 strains were observed in 2009/2010, most likely induced by the start of the 2009 pandemic, and the subsequent post-pandemic seasons from 2010/2011 onward when A/California/07/2009 became the predominant strain. In the 2014/2015 season, population immunity against A/California/07/2009 and A/Michigan/45/2015 strains increased again, associated with strong cross-reactions. Whereas hemagglutination inhibition assay has a higher sensitivity for detection of new seasonal drift, the single radial hemolysis assay is an excellent tool for determining the presence of pre-existing immunity, allowing a potential prediction on the booster potential of influenza vaccines against newly emerging drifted strains.

ACS Style

Serena Marchi; Ilaria Manini; Otfried Kistner; Pietro Piu; Edmond J. Remarque; Alessandro Manenti; Fabrizio Biuso; Tommaso Carli; Giacomo Lazzeri; Emanuele Montomoli; Claudia Maria Trombetta. Serologically-Based Evaluation of Cross-Protection Antibody Responses among Different A(H1N1) Influenza Strains. Vaccines 2020, 8, 656 .

AMA Style

Serena Marchi, Ilaria Manini, Otfried Kistner, Pietro Piu, Edmond J. Remarque, Alessandro Manenti, Fabrizio Biuso, Tommaso Carli, Giacomo Lazzeri, Emanuele Montomoli, Claudia Maria Trombetta. Serologically-Based Evaluation of Cross-Protection Antibody Responses among Different A(H1N1) Influenza Strains. Vaccines. 2020; 8 (4):656.

Chicago/Turabian Style

Serena Marchi; Ilaria Manini; Otfried Kistner; Pietro Piu; Edmond J. Remarque; Alessandro Manenti; Fabrizio Biuso; Tommaso Carli; Giacomo Lazzeri; Emanuele Montomoli; Claudia Maria Trombetta. 2020. "Serologically-Based Evaluation of Cross-Protection Antibody Responses among Different A(H1N1) Influenza Strains." Vaccines 8, no. 4: 656.

Preprint
Published: 05 October 2020
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After the influenza H1N1 pandemic of 2009, the seasonal A/Brisbane/59/2007 strain was replaced by the A/California/07/2009 strain for the influenza virus vaccine composition. After several seasons with no indications on the occurrence of antigenic drift, A/Michigan/45/2015 was chosen as the H1N1 vaccine strain for season 2017/2018. Since the immune response to influenza is shaped by the history of exposure to antigenically similar strains, the potential cross-protection between seasonal human influenza vaccine strains and the emerging pandemic strains was investigated. Human serum samples were tested by haemagglutination inhibition and single radial haemolysis assays against A/Brisbane/59/2007, A/California/07/2009, and A/Michigan/45/2015 strains. Strong cross-reactions between A/California/07/2009 and A/Michigan/45/2015 strains were observed in 2009/2010, most likely induced by the start of the 2009 pandemic, and the subsequent post-pandemic seasons from 2010/2011 onwards when A/California/07/2009 becomes the predominant strain. In 2014/2015 season, population immunity against A/California/07/2009 and A/Michigan/45/2015 strains increased again, associated with strong cross-reactions. While haemagglutination inhibition assay has a higher sensitivity for detection of new seasonal drift, the single radial haemolysis assay is an excellent tool to determine the presence of pre-existing immunity, allowing a potential prediction on the booster potential of influenza vaccines against newly emerging drifted strains.

ACS Style

Serena Marchi; Ilaria Manini; Otfried Kistner; Pietro Piu; Edmond J. Remarque; Alessandro Manenti; Fabrizio Biuso; Tommaso Carli; Giacomo Lazzeri; Emanuele Montomoli; Claudia Maria Trombetta. Serologically-based Evaluation of Cross-protection Antibody Responses Among Different A(H1N1) Influenza Strains. 2020, 1 .

AMA Style

Serena Marchi, Ilaria Manini, Otfried Kistner, Pietro Piu, Edmond J. Remarque, Alessandro Manenti, Fabrizio Biuso, Tommaso Carli, Giacomo Lazzeri, Emanuele Montomoli, Claudia Maria Trombetta. Serologically-based Evaluation of Cross-protection Antibody Responses Among Different A(H1N1) Influenza Strains. . 2020; ():1.

Chicago/Turabian Style

Serena Marchi; Ilaria Manini; Otfried Kistner; Pietro Piu; Edmond J. Remarque; Alessandro Manenti; Fabrizio Biuso; Tommaso Carli; Giacomo Lazzeri; Emanuele Montomoli; Claudia Maria Trombetta. 2020. "Serologically-based Evaluation of Cross-protection Antibody Responses Among Different A(H1N1) Influenza Strains." , no. : 1.

Journal article
Published: 11 September 2020 in Journal of Public Health
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Background Despite efforts to increase coverage by two doses of measles vaccine in Italy, measles continues to circulate, with over 13 000 cases of disease since 2013. This study aimed to evaluate immunity to measles in Italian children and adolescents. Methods A total of 378 serum samples from subjects aged 9 months–18 years were collected in Northern, Central and Southern regions of Italy between 2012 and 2016. Specific IgG antibodies against measles were measured by a commercial ELISA kit. Results The frequency of IgG-positive samples ranged from 10.5% in infants under 1 year to 98.3% in children aged 6–7 years. The frequency of IgG was 72.2% in subjects aged 1–2 years, 85.6% in those aged 3–5 years and 88.3 and 86.8% in those aged 8–10 and 11–18 years, respectively. In Northern Italy, IgG prevalence was consistent with data on vaccination coverage, whereas some differences were observed in samples from subjects aged more than 8 years in Central and Southern Italy. Conclusions Our findings confirm that a large proportion of children and adolescents in Italy are still susceptible to measles. While data on first- and second-dose measles vaccination are essential, they are not sufficient to identify susceptible population cohorts to be targeted by vaccination.

ACS Style

Serena Marchi; Martina Monti; Simonetta Viviani; Edmond J Remarque; Susanna Esposito; Nicola Principi; Luca Ruggiero; Emanuele Montomoli; Claudia Maria Trombetta. Immunity to measles in Italian children and adolescents: a persistent problem in view of measles elimination. Journal of Public Health 2020, 1 .

AMA Style

Serena Marchi, Martina Monti, Simonetta Viviani, Edmond J Remarque, Susanna Esposito, Nicola Principi, Luca Ruggiero, Emanuele Montomoli, Claudia Maria Trombetta. Immunity to measles in Italian children and adolescents: a persistent problem in view of measles elimination. Journal of Public Health. 2020; ():1.

Chicago/Turabian Style

Serena Marchi; Martina Monti; Simonetta Viviani; Edmond J Remarque; Susanna Esposito; Nicola Principi; Luca Ruggiero; Emanuele Montomoli; Claudia Maria Trombetta. 2020. "Immunity to measles in Italian children and adolescents: a persistent problem in view of measles elimination." Journal of Public Health , no. : 1.

Preprint content
Published: 10 August 2020
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A newly identified coronavirus, named SARS-CoV-2, emerged in December 2019 in Hubei Province, China, and quickly spread throughout the world; so far, it has caused more than 18 million cases of disease and 700,000 deaths. The diagnosis of SARS-CoV-2 infection is currently based on the detection of viral RNA in nasopharyngeal swabs by means of molecular-based assays, such as real-time RT-PCR. Furthermore, serological assays aimed at detecting different classes of antibodies constitute the best surveillance strategy for gathering information on the humoral immune response to infection and the spread of the virus through the population, in order to evaluate the immunogenicity of novel future vaccines and medicines for the treatment and prevention of COVID-19 disease. The aim of this study was to determine SARS-CoV-2-specific antibodies in human serum samples by means of different commercial and in-house ELISA kits, in order to evaluate and compare their results first with one another and then with those yielded by functional assays using wild-type virus. It is important to know the level of SARS-CoV-2-specific IgM, IgG and IgA antibodies in order to predict population immunity and possible cross-reactivity with other coronaviruses and to identify potentially infectious subjects. In addition, in a small sub-group of samples, we performed a subtyping Immunoglobulin G ELISA. Our data showed an excellent statistical correlation between the neutralization titer and the IgG, IgM and IgA ELISA response against the receptor-binding domain of the spike protein, confirming that antibodies against this portion of the virus spike protein are highly neutralizing and that the ELISA Receptor-Binding Domain-based assay can be used as a valid surrogate for the neutralization assay in laboratories which do not have Biosecurity level-3 facilities.

ACS Style

Livia Mazzini; Donata Martinuzzi; Inesa Hyseni; Giulia Lapini; Linda Benincasa; Pietro Piu; Claudia Maria Trombetta; Serena Marchi; Ilaria Razzano; Alessandro Manenti; Emanuele Montomoli. Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples. 2020, 1 .

AMA Style

Livia Mazzini, Donata Martinuzzi, Inesa Hyseni, Giulia Lapini, Linda Benincasa, Pietro Piu, Claudia Maria Trombetta, Serena Marchi, Ilaria Razzano, Alessandro Manenti, Emanuele Montomoli. Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples. . 2020; ():1.

Chicago/Turabian Style

Livia Mazzini; Donata Martinuzzi; Inesa Hyseni; Giulia Lapini; Linda Benincasa; Pietro Piu; Claudia Maria Trombetta; Serena Marchi; Ilaria Razzano; Alessandro Manenti; Emanuele Montomoli. 2020. "Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples." , no. : 1.

Abstract
Published: 24 June 2020 in Proceedings
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Before its recent spread, serological investigations conducted between the 1960s and the 1990s showed the wide presence of Zika virus in Africa. According to the World Health Organization, the entire Africa continent is at risk of Zika outbreak due to the presence of the virus, competent vectors, and the low capacity for surveillance and containment of an epidemic. However, limited data are available on the recent prevalence in the African population. The aim of this study was to evaluate the immunity against Zika virus in samples of a selected cohort from West Africa, in order to investigate the circulation of the virus in the region during the first years of its emergence in the Pacific. Human serum samples were collected in 2007 and between 2011 and 2012 from a cohort of subjects from Mali, Senegal, and The Gambia. The samples were tested using an enzyme-linked immunosorbent assay (ELISA) detection kit and positives were further confirmed by microneutralization test. The results indicate that Zika virus is present and actively circulating in Senegal and The Gambia, with prevalence values of 13.7% and 6.9% in 2012, respectively. Although no significant differences in prevalence were found for the considered time period, seroconversion of some subjects showed the active circulation of Zika virus in the West African area. Analysis by age showed an increase in immunity in relation to increasing age, demonstrating that the population is consistently exposed to the virus throughout life and with a high possibility of being infected during reproductive age. In conclusion, the obtained results allow for better knowledge of the circulation of Zika virus within three different ecological and demographic contexts, and represent an update to the limited data currently available.

ACS Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Yuxiao Tang; Adele Boccuto; Ilaria Vicenti; Maurizio Zazzi; Samba Sow; Aldiouma Diallo; Olubukola T. Idoko; Niranjan Bhat; Claudia Maria Trombetta. Zika Virus Epidemiology in Selected West African Countries between 2007 and 2012. Proceedings 2020, 50, 100 .

AMA Style

Serena Marchi, Simonetta Viviani, Emanuele Montomoli, Yuxiao Tang, Adele Boccuto, Ilaria Vicenti, Maurizio Zazzi, Samba Sow, Aldiouma Diallo, Olubukola T. Idoko, Niranjan Bhat, Claudia Maria Trombetta. Zika Virus Epidemiology in Selected West African Countries between 2007 and 2012. Proceedings. 2020; 50 (1):100.

Chicago/Turabian Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Yuxiao Tang; Adele Boccuto; Ilaria Vicenti; Maurizio Zazzi; Samba Sow; Aldiouma Diallo; Olubukola T. Idoko; Niranjan Bhat; Claudia Maria Trombetta. 2020. "Zika Virus Epidemiology in Selected West African Countries between 2007 and 2012." Proceedings 50, no. 1: 100.

Journal article
Published: 13 June 2020 in Viruses
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According to the World Health Organization, the entire African continent is at risk of a Zika outbreak. To increase data availability on the epidemiology of Zika virus circulation in Africa, we evaluated the immunity to Zika virus in a selected cohort of subjects from West Africa between 2007 and 2012. Human serum samples were collected in 2007 and in 2011/2012 from a cohort of 2–29-year-old subjects from Mali, Senegal, and The Gambia. A sample that tested positive by Zika virus IgG ELISA and by Zika virus microneutralization test was defined as positive. In 2007, the highest prevalence was 21.9%, found in Senegal among 18–29-year-old subjects. In 2011/2012, the highest prevalence, 22.7%, was found still in Senegal, but in 11–17-year-old subjects. During both study periods, the lowest prevalence was found in Mali, where few positive cases were found only in 18–29-year-old subjects. The Gambia showed an intermediate prevalence. In the three countries, prevalence was strongly associated with increasing age. This study contributes to understanding Zika virus circulation within three different ecological and demographic contexts with scarce or no data currently available. Results showed that Zika virus circulated actively in West Africa between the period 2007 and 2011/2012, but with some geographic specificity.

ACS Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Yuxiao Tang; Adele Boccuto; Ilaria Vicenti; Maurizio Zazzi; Samba Sow; Aldiouma Diallo; Olubukola T. Idoko; Niranjan Bhat; Claudia Maria Trombetta. Zika Virus in West Africa: A Seroepidemiological Study between 2007 and 2012. Viruses 2020, 12, 1 .

AMA Style

Serena Marchi, Simonetta Viviani, Emanuele Montomoli, Yuxiao Tang, Adele Boccuto, Ilaria Vicenti, Maurizio Zazzi, Samba Sow, Aldiouma Diallo, Olubukola T. Idoko, Niranjan Bhat, Claudia Maria Trombetta. Zika Virus in West Africa: A Seroepidemiological Study between 2007 and 2012. Viruses. 2020; 12 (6):1.

Chicago/Turabian Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Yuxiao Tang; Adele Boccuto; Ilaria Vicenti; Maurizio Zazzi; Samba Sow; Aldiouma Diallo; Olubukola T. Idoko; Niranjan Bhat; Claudia Maria Trombetta. 2020. "Zika Virus in West Africa: A Seroepidemiological Study between 2007 and 2012." Viruses 12, no. 6: 1.

Letter to the editor
Published: 08 June 2020 in European Journal of Internal Medicine
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At the end of December 2019, the first cases of SARS-CoV-2 infection were identified in Wuhan, China. [1World Health Organization (WHO). WHO director-general's opening remarks at the media briefing on COVID-19: 11 March 2020. Published March 11, 2020. Accessed March 11, 2020. https://www.who.int/dg/speeches/detail/who-director-general-s-opening-remarks-at-the-media-briefing-on-covid-19—11-march-2020.Google Scholar] In the following months, the outbreak of SARS-COV-2 infections spreads worldwide [1World Health Organization (WHO). WHO director-general's opening remarks at the media briefing on COVID-19: 11 March 2020. Published March 11, 2020. Accessed March 11, 2020. https://www.who.int/dg/speeches/detail/who-director-general-s-opening-remarks-at-the-media-briefing-on-covid-19—11-march-2020.Google Scholar] As of May 1, 2020, more than 3 million of SARS-CoV-2 infections have been detected worldwide. These data were mainly collected from inpatients. On the other hand, asymptomatic or mildly symptomatic subjects are usually untested for SARS-CoV-2 although they are supposed to far outnumber the symptomatic patients. [2Day M Covid-19: four fifths of cases are asymptomatic, China figures indicate.BMJ. 2020; 369: m1375Crossref PubMed Scopus (129) Google Scholar] Therefore, there is an urgent need to estimate as accurately as possible the real number of subjects who have been infected by the virus. The aim of this study was to assess the frequency of asymptomatic subjects with a SARS-CoV-2 positive nasal swab or presenting immunoglobulins against the virus in Milan (Italy).

ACS Style

Gregorio Paolo Milani; Emanuele Montomoli; Valentina Bollati; Benedetta Albetti; Claudio Bandi; Tommaso Bellini; Matteo Bonzini; Marco Buscaglia; Carlo Cantarella; Laura Cantone; Michele Carugno; Sergio Casartelli; Guido Cavaletti; Sarah D'Alessandro; Francesca De Chiara; Serena Delbue; Laura Dioni; Ivano Eberini; Chiara Favero; Luca Ferrari; Monica Ferraroni; Laura Galastri; Cristina Galli; Mirjam Hoxha; Simona Iodice; Carlo La Vecchia; Chiara Macchi; Ilaria Manini; Serena Marchi; Jacopo Mariani; Elena Pariani; Angela Cecilia Pesatori; Federica Rota; Massimiliano Ruscica; Tommaso Schioppo; Letizia Tarantini; Claudia Maria Trombetta; Maria Grazia Valsecchi; Marco Vicenzi; Giuliano Zanchetta. SARS-CoV-2 infection among asymptomatic homebound subjects in Milan, Italy. European Journal of Internal Medicine 2020, 78, 161 -163.

AMA Style

Gregorio Paolo Milani, Emanuele Montomoli, Valentina Bollati, Benedetta Albetti, Claudio Bandi, Tommaso Bellini, Matteo Bonzini, Marco Buscaglia, Carlo Cantarella, Laura Cantone, Michele Carugno, Sergio Casartelli, Guido Cavaletti, Sarah D'Alessandro, Francesca De Chiara, Serena Delbue, Laura Dioni, Ivano Eberini, Chiara Favero, Luca Ferrari, Monica Ferraroni, Laura Galastri, Cristina Galli, Mirjam Hoxha, Simona Iodice, Carlo La Vecchia, Chiara Macchi, Ilaria Manini, Serena Marchi, Jacopo Mariani, Elena Pariani, Angela Cecilia Pesatori, Federica Rota, Massimiliano Ruscica, Tommaso Schioppo, Letizia Tarantini, Claudia Maria Trombetta, Maria Grazia Valsecchi, Marco Vicenzi, Giuliano Zanchetta. SARS-CoV-2 infection among asymptomatic homebound subjects in Milan, Italy. European Journal of Internal Medicine. 2020; 78 ():161-163.

Chicago/Turabian Style

Gregorio Paolo Milani; Emanuele Montomoli; Valentina Bollati; Benedetta Albetti; Claudio Bandi; Tommaso Bellini; Matteo Bonzini; Marco Buscaglia; Carlo Cantarella; Laura Cantone; Michele Carugno; Sergio Casartelli; Guido Cavaletti; Sarah D'Alessandro; Francesca De Chiara; Serena Delbue; Laura Dioni; Ivano Eberini; Chiara Favero; Luca Ferrari; Monica Ferraroni; Laura Galastri; Cristina Galli; Mirjam Hoxha; Simona Iodice; Carlo La Vecchia; Chiara Macchi; Ilaria Manini; Serena Marchi; Jacopo Mariani; Elena Pariani; Angela Cecilia Pesatori; Federica Rota; Massimiliano Ruscica; Tommaso Schioppo; Letizia Tarantini; Claudia Maria Trombetta; Maria Grazia Valsecchi; Marco Vicenzi; Giuliano Zanchetta. 2020. "SARS-CoV-2 infection among asymptomatic homebound subjects in Milan, Italy." European Journal of Internal Medicine 78, no. : 161-163.

Journal article
Published: 10 May 2020 in Vaccines
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Varicella is a highly contagious, infectious disease caused by the varicella-zoster virus. Those at higher risk of severe complications are immunocompromised individuals, adults, non-immune pregnant women, and newborns. According to the gestational time, when varicella-zoster virus infection is acquired during pregnancy, serious complications can potentially occur for both the woman and the fetus. The present study was conducted to assess the profile of varicella susceptibility in pregnant women in Apulia, a large region in Southern Italy, from 2016 to 2019. The data showed that pregnant women between the age of 15–24 and 40–49 years, the youngest and the oldest, respectively, are the most protected against varicella-zoster virus infection, exceeding the prevalence rate of 90%. Conversely, pregnant women between the age of 25 and 34 years seem to be the most vulnerable and the most at risk for acquiring varicella-zoster virus infection during pregnancy. Analysis of the immunity status against varicella should be introduced as a screening test before pregnancy, together with a strategic vaccination campaign targeting non-immune women of childbearing age, in order to reduce the risk of congenital and perinatal varicella.

ACS Style

Claudia M. Trombetta; Emanuele Montomoli; Simonetta Viviani; Rosa Coluccio; Serena Marchi. Evaluation of Varicella Immunity during Pregnancy in Apulia Region, Southern Italy. Vaccines 2020, 8, 1 .

AMA Style

Claudia M. Trombetta, Emanuele Montomoli, Simonetta Viviani, Rosa Coluccio, Serena Marchi. Evaluation of Varicella Immunity during Pregnancy in Apulia Region, Southern Italy. Vaccines. 2020; 8 (2):1.

Chicago/Turabian Style

Claudia M. Trombetta; Emanuele Montomoli; Simonetta Viviani; Rosa Coluccio; Serena Marchi. 2020. "Evaluation of Varicella Immunity during Pregnancy in Apulia Region, Southern Italy." Vaccines 8, no. 2: 1.

Research article
Published: 08 May 2020 in Journal of Medical Virology
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The micro‐neutralization assay is a fundamental test in virology, immunology, vaccine assessment and epidemiology studies. Since the SARS‐CoV‐2 outbreak at the end of December 2019 in China, it has become extremely important to have well‐established and validated diagnostic and serological assays for this new emerging virus. Here, we present a micro‐neutralization assay with the use of SARS‐CoV‐2 wild type virus with two different methods of read‐out. We evaluated the performance of this assay using human serum samples taken from an italian seroepidemiological study being performed at the University of Siena, along with the human monoclonal antibody CR3022 and some iper‐immune animal serum samples against Influenza and Adenovirus strains. The same panel of human samples have been previously tested in ELISA as a pre‐screening. Positive, borderline and negative ELISA samples were evaluated in neutralization assay using two different method of read‐out: subjective (by means of an inverted optical microscope) and objective (by means of a spectrophotometer). Our findings suggest that at least 50% of positive ELISA samples are positive in neutralization as well, and that method is able to quantify different antibody concentrations in a specific manner. Taken together, our results confirm that the colorimetric cytopathic effect‐based microneutralization assay could be used as a valid clinical test method for epidemiological and vaccines studies. This article is protected by copyright. All rights reserved.

ACS Style

Alessandro Manenti; Marta Maggetti; Elisa Casa; Donata Martinuzzi; Alessandro Torelli; Claudia M. Trombetta; Serena Marchi; Emanuele Montomoli. Evaluation of SARS‐CoV‐2 neutralizing antibodies using a CPE‐based colorimetric live virus micro‐neutralization assay in human serum samples. Journal of Medical Virology 2020, 92, 2096 -2104.

AMA Style

Alessandro Manenti, Marta Maggetti, Elisa Casa, Donata Martinuzzi, Alessandro Torelli, Claudia M. Trombetta, Serena Marchi, Emanuele Montomoli. Evaluation of SARS‐CoV‐2 neutralizing antibodies using a CPE‐based colorimetric live virus micro‐neutralization assay in human serum samples. Journal of Medical Virology. 2020; 92 (10):2096-2104.

Chicago/Turabian Style

Alessandro Manenti; Marta Maggetti; Elisa Casa; Donata Martinuzzi; Alessandro Torelli; Claudia M. Trombetta; Serena Marchi; Emanuele Montomoli. 2020. "Evaluation of SARS‐CoV‐2 neutralizing antibodies using a CPE‐based colorimetric live virus micro‐neutralization assay in human serum samples." Journal of Medical Virology 92, no. 10: 2096-2104.

Journal article
Published: 28 April 2020 in Journal of Infectious Diseases
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ACS Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Claudia Maria Trombetta. Measles Susceptibility in Apulia: The Italian Mirror? Journal of Infectious Diseases 2020, 222, 1414 -1415.

AMA Style

Serena Marchi, Simonetta Viviani, Emanuele Montomoli, Claudia Maria Trombetta. Measles Susceptibility in Apulia: The Italian Mirror? Journal of Infectious Diseases. 2020; 222 (8):1414-1415.

Chicago/Turabian Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Claudia Maria Trombetta. 2020. "Measles Susceptibility in Apulia: The Italian Mirror?" Journal of Infectious Diseases 222, no. 8: 1414-1415.

Rapid communication
Published: 09 April 2020 in ACS Medicinal Chemistry Letters
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In the absence of effective drugs or vaccines for the treatment of the five Dengue Virus serotypes, the search for novel antiviral drugs is of primary importance for the scientific community. In this context, drug repurposing represents the most used strategy; however, the study of host targets is now attracting attention since it allows identification of broad-spectrum drugs endowed with high genetic barrier. In the last ten years our research group identified several small molecules DDX3X inhibitors and proved their efficacy against different viruses including novel emerging ones. Herein, starting from a screening of our compounds, we designed and synthesized novel derivatives with potent activity and high selectivity. Finally, we synthesized a fluorescent inhibitor that allowed us to study DDX3X cellular localization during DENV infection in vitro. Immunofluorescence analysis showed that our inhibitor colocalized with DDX3X, promoting the reduction of infected cells and recovering the number of viable cells.

ACS Style

Annalaura Brai; Adele Boccuto; Martina Monti; Serena Marchi; Ilaria Vicenti; Francesco Saladini; Claudia Immacolata Trivisani; Alessandro Pollutri; Claudia Maria Trombetta; Emanuele Montomoli; Valentina Riva; Anna Garbelli; Emanuele Maria Nola; Maurizio Zazzi; Giovanni Maga; Elena Dreassi; Maurizio Botta. Exploring the Implication of DDX3X in DENV Infection: Discovery of the First-in-Class DDX3X Fluorescent Inhibitor. ACS Medicinal Chemistry Letters 2020, 11, 956 -962.

AMA Style

Annalaura Brai, Adele Boccuto, Martina Monti, Serena Marchi, Ilaria Vicenti, Francesco Saladini, Claudia Immacolata Trivisani, Alessandro Pollutri, Claudia Maria Trombetta, Emanuele Montomoli, Valentina Riva, Anna Garbelli, Emanuele Maria Nola, Maurizio Zazzi, Giovanni Maga, Elena Dreassi, Maurizio Botta. Exploring the Implication of DDX3X in DENV Infection: Discovery of the First-in-Class DDX3X Fluorescent Inhibitor. ACS Medicinal Chemistry Letters. 2020; 11 (5):956-962.

Chicago/Turabian Style

Annalaura Brai; Adele Boccuto; Martina Monti; Serena Marchi; Ilaria Vicenti; Francesco Saladini; Claudia Immacolata Trivisani; Alessandro Pollutri; Claudia Maria Trombetta; Emanuele Montomoli; Valentina Riva; Anna Garbelli; Emanuele Maria Nola; Maurizio Zazzi; Giovanni Maga; Elena Dreassi; Maurizio Botta. 2020. "Exploring the Implication of DDX3X in DENV Infection: Discovery of the First-in-Class DDX3X Fluorescent Inhibitor." ACS Medicinal Chemistry Letters 11, no. 5: 956-962.

Brief report
Published: 14 January 2020 in Parasite
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Toxoplasmosis is a worldwide health problem. Infection in pregnant women can result in severe fetal morbidity or in subclinical neonatal infection; most subclinical cases develop ocular and neurological sequelae. The purpose of this serological study was to assess the prevalence of Toxoplasma gondii in two populations of women of childbearing age in Siena (Tuscany, Central Italy) and Bari (Apulia, Southern Italy) between 2013 and 2017 and in a group of pregnant women in Bari in 2016–2017. Serum samples were tested for the presence of specific anti-Toxoplasma gondii IgG antibodies by a commercially available ELISA test. The percentage of seropositive subjects in Bari was significantly higher than in Siena (22.4% vs. 12.4%) and an age-related trend was observed. A low prevalence of T. gondii infection (13.8%) was observed among the pregnant women tested. In addition to showing a significant difference between Central and Southern Italy, this study provides updated data on T. gondii seroprevalence in women during childbearing age and pregnancy. The results confirm a trend toward a decrease, especially in younger people and pregnant women.

ACS Style

Daniela Fanigliulo; Serena Marchi; Emanuele Montomoli; Claudia Maria Trombetta. Toxoplasma gondii in women of childbearing age and during pregnancy: seroprevalence study in Central and Southern Italy from 2013 to 2017. Parasite 2020, 27, 2 .

AMA Style

Daniela Fanigliulo, Serena Marchi, Emanuele Montomoli, Claudia Maria Trombetta. Toxoplasma gondii in women of childbearing age and during pregnancy: seroprevalence study in Central and Southern Italy from 2013 to 2017. Parasite. 2020; 27 ():2.

Chicago/Turabian Style

Daniela Fanigliulo; Serena Marchi; Emanuele Montomoli; Claudia Maria Trombetta. 2020. "Toxoplasma gondii in women of childbearing age and during pregnancy: seroprevalence study in Central and Southern Italy from 2013 to 2017." Parasite 27, no. : 2.

Conference paper
Published: 01 January 2020 in Proceedings
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Influenza D virus (IDV) is a novel influenza virus first isolated from swine in 2011 in Oklahoma. Several studies have isolated IDV in cattle from multiple geographic areas, suggesting that cattle could be a possible primary natural reservoir for the virus. To date, few studies have been performed on human samples and there is no conclusive evidence that IDV has the ability to infect humans. This serological study aimed to assess the prevalence of antibodies against IDV in the human population. The IDV used in the serological analysis was influenza D/bovine/Oklahoma/660/2013. The human serum samples, collected in Italy between 2005 and 2017, were randomly selected from the laboratory serum bank and tested by the haemagglutination inhibition (HI) assay. HI positivity has been confirmed using the virus neutralization (VN) assay. Based on HI positivity (HI titers ≥ 10), a low prevalence (5%–10%) was observed between 2005 and 2007. There has been a sharp increase since 2008, resulting in two main peaks in 2009–2010 and 2013–2014, a finding confirmed by the statistical trend analysis. The same pattern and trends can be seen with higher HI titers of >20 and ≥40. The prevalence of antibodies against IDV has increased in the human population in Italy from 2005 to 2017. Low prevalence values between 2005 and 2007 suggest that IDV most probably circulated before its detection in 2011, and perhaps even before 2005. In Italy, IDV has been shown to circulate among swine and bovine herds. It is, therefore, possible that prevalence peaks in humans follow the infection epidemics in animals and do not to persist in the population, resembling a spillover event from the animal reservoir and showing that the virus may not circulate consistently in the human population. However, IDV seemed to have the ability to elicit an immune response in humans.

ACS Style

Claudia Maria Trombetta; Serena Marchi; Ilaria Manini; Otfried Kistner; Feng Li; Pietro Piu; Alessandro Manenti; Fabrizio Biuso; Chithra Sreenivasan; Julian Druce; Emanuele Montomoli. Influenza D Virus: A Potential Threat for Humans? Proceedings 2020, 50, 1 .

AMA Style

Claudia Maria Trombetta, Serena Marchi, Ilaria Manini, Otfried Kistner, Feng Li, Pietro Piu, Alessandro Manenti, Fabrizio Biuso, Chithra Sreenivasan, Julian Druce, Emanuele Montomoli. Influenza D Virus: A Potential Threat for Humans? Proceedings. 2020; 50 (1):1.

Chicago/Turabian Style

Claudia Maria Trombetta; Serena Marchi; Ilaria Manini; Otfried Kistner; Feng Li; Pietro Piu; Alessandro Manenti; Fabrizio Biuso; Chithra Sreenivasan; Julian Druce; Emanuele Montomoli. 2020. "Influenza D Virus: A Potential Threat for Humans?" Proceedings 50, no. 1: 1.

Journal article
Published: 27 December 2019 in Viruses
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Influenza D virus is a novel influenza virus, which was first isolated from an ailing swine in 2011 and later detected in cattle, suggesting that these animals may be a primary natural reservoir. To date, few studies have been performed on human samples and there is no conclusive evidence on the ability of the virus to infect humans. The aim of this serological study was to assess the prevalence of antibodies against influenza D virus in human serum samples collected in Italy from 2005 to 2017. Serum samples were analysed by haemagglutination inhibition and virus neutralization assays. The results showed that the prevalence of antibodies against the virus increased in the human population in Italy from 2005 to 2017, with a trend characterized by a sharp increase in some years, followed by a decline in subsequent years. The virus showed the ability to infect and elicit an immune response in humans. However, prevalence peaks in humans appear to follow epidemics in animals and not to persist in the human population.

ACS Style

Claudia M. Trombetta; Serena Marchi; Ilaria Manini; Otfried Kistner; Feng Li; Pietro Piu; Alessandro Manenti; Fabrizio Biuso; Chithra Sreenivasan; Julian Druce; Emanuele Montomoli. Influenza D Virus: Serological Evidence in the Italian Population from 2005 to 2017. Viruses 2019, 12, 30 .

AMA Style

Claudia M. Trombetta, Serena Marchi, Ilaria Manini, Otfried Kistner, Feng Li, Pietro Piu, Alessandro Manenti, Fabrizio Biuso, Chithra Sreenivasan, Julian Druce, Emanuele Montomoli. Influenza D Virus: Serological Evidence in the Italian Population from 2005 to 2017. Viruses. 2019; 12 (1):30.

Chicago/Turabian Style

Claudia M. Trombetta; Serena Marchi; Ilaria Manini; Otfried Kistner; Feng Li; Pietro Piu; Alessandro Manenti; Fabrizio Biuso; Chithra Sreenivasan; Julian Druce; Emanuele Montomoli. 2019. "Influenza D Virus: Serological Evidence in the Italian Population from 2005 to 2017." Viruses 12, no. 1: 30.

Short report
Published: 26 November 2019 in Human Vaccines & Immunotherapeutics
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Prevention of congenital rubella is achieved by vaccination of susceptible women of childbearing age. In Italy, the National Plan for Measles and Congenital Rubella Elimination 2010–2015 implemented catch-up vaccination activities targeting susceptible adolescents and young adults, including women of childbearing age. The aim of this study was to assess the immunity against rubella in women of childbearing age in Tuscany (Central Italy) and Apulia (Southern Italy) and pregnant women in Apulia after the implementation of the National Plan for Measles and Congenital Rubella Elimination. Overall, anti-rubella IgG prevalence in women of childbearing age samples was 88.6% in Tuscany and 84.3% in Apulia. The lowest prevalence was observed in samples of 26–35 years old women of childbearing age in Apulia with 77.8%. Only 62.7% of samples from 26–35 years old pregnant women had IgG against rubella, and one sample out of 95 was positive to IgM. The findings of this study highlight the need for increasing awareness on the risk of contracting rubella infection during pregnancy and implement vaccination strategies to create opportunities for administration of rubella containing vaccines in young girls and women of childbearing age.

ACS Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Claudia Maria Trombetta. Elimination of congenital rubella: a seroprevalence study of pregnant women and women of childbearing age in Italy. Human Vaccines & Immunotherapeutics 2019, 16, 895 -898.

AMA Style

Serena Marchi, Simonetta Viviani, Emanuele Montomoli, Claudia Maria Trombetta. Elimination of congenital rubella: a seroprevalence study of pregnant women and women of childbearing age in Italy. Human Vaccines & Immunotherapeutics. 2019; 16 (4):895-898.

Chicago/Turabian Style

Serena Marchi; Simonetta Viviani; Emanuele Montomoli; Claudia Maria Trombetta. 2019. "Elimination of congenital rubella: a seroprevalence study of pregnant women and women of childbearing age in Italy." Human Vaccines & Immunotherapeutics 16, no. 4: 895-898.