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Roman Wölfel
Bundeswehr Institute of Microbiology, 80937 Munich, Germany

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Journal article
Published: 30 March 2021 in International Journal of Environmental Research and Public Health
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Given the large number of mild or asymptomatic SARS-CoV-2 cases, only population-based studies can provide reliable estimates of the magnitude of the pandemic. We therefore aimed to assess the sero-prevalence of SARS-CoV-2 in the Munich general population after the first wave of the pandemic. For this purpose, we drew a representative sample of 2994 private households and invited household members 14 years and older to complete questionnaires and to provide blood samples. SARS-CoV-2 seropositivity was defined as Roche N pan-Ig ≥ 0.4218. We adjusted the prevalence for the sampling design, sensitivity, and specificity. We investigated risk factors for SARS-CoV-2 seropositivity and geospatial transmission patterns by generalized linear mixed models and permutation tests. Seropositivity for SARS-CoV-2-specific antibodies was 1.82% (95% confidence interval (CI) 1.28–2.37%) as compared to 0.46% PCR-positive cases officially registered in Munich. Loss of the sense of smell or taste was associated with seropositivity (odds ratio (OR) 47.4; 95% CI 7.2–307.0) and infections clustered within households. By this first population-based study on SARS-CoV-2 prevalence in a large German municipality not affected by a superspreading event, we could show that at least one in four cases in private households was reported and known to the health authorities. These results will help authorities to estimate the true burden of disease in the population and to take evidence-based decisions on public health measures.

ACS Style

Michael Pritsch; Katja Radon; Abhishek Bakuli; Ronan Le Gleut; Laura Olbrich; Jessica Guggenbüehl Noller; Elmar Saathoff; Noemi Castelletti; Mercè Garí; Peter Pütz; Yannik Schälte; Turid Frahnow; Roman Wölfel; Camilla Rothe; Michel Pletschette; Dafni Metaxa; Felix Forster; Verena Thiel; Friedrich Rieß; Maximilian Diefenbach; Günter Fröschl; Jan Bruger; Simon Winter; Jonathan Frese; Kerstin Puchinger; Isabel Brand; Inge Kroidl; Jan Hasenauer; Christiane Fuchs; Andreas Wieser; Michael Hoelscher; on behalf of the KoCo19 study group. Prevalence and Risk Factors of Infection in the Representative COVID-19 Cohort Munich. International Journal of Environmental Research and Public Health 2021, 18, 3572 .

AMA Style

Michael Pritsch, Katja Radon, Abhishek Bakuli, Ronan Le Gleut, Laura Olbrich, Jessica Guggenbüehl Noller, Elmar Saathoff, Noemi Castelletti, Mercè Garí, Peter Pütz, Yannik Schälte, Turid Frahnow, Roman Wölfel, Camilla Rothe, Michel Pletschette, Dafni Metaxa, Felix Forster, Verena Thiel, Friedrich Rieß, Maximilian Diefenbach, Günter Fröschl, Jan Bruger, Simon Winter, Jonathan Frese, Kerstin Puchinger, Isabel Brand, Inge Kroidl, Jan Hasenauer, Christiane Fuchs, Andreas Wieser, Michael Hoelscher, on behalf of the KoCo19 study group. Prevalence and Risk Factors of Infection in the Representative COVID-19 Cohort Munich. International Journal of Environmental Research and Public Health. 2021; 18 (7):3572.

Chicago/Turabian Style

Michael Pritsch; Katja Radon; Abhishek Bakuli; Ronan Le Gleut; Laura Olbrich; Jessica Guggenbüehl Noller; Elmar Saathoff; Noemi Castelletti; Mercè Garí; Peter Pütz; Yannik Schälte; Turid Frahnow; Roman Wölfel; Camilla Rothe; Michel Pletschette; Dafni Metaxa; Felix Forster; Verena Thiel; Friedrich Rieß; Maximilian Diefenbach; Günter Fröschl; Jan Bruger; Simon Winter; Jonathan Frese; Kerstin Puchinger; Isabel Brand; Inge Kroidl; Jan Hasenauer; Christiane Fuchs; Andreas Wieser; Michael Hoelscher; on behalf of the KoCo19 study group. 2021. "Prevalence and Risk Factors of Infection in the Representative COVID-19 Cohort Munich." International Journal of Environmental Research and Public Health 18, no. 7: 3572.

Comparative study
Published: 08 March 2021 in Journal of Virological Methods
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Reliable methods for the detection of SARS-CoV-2 neutralising antibodies (NAbs) are essential for the evaluation of vaccine candidates and for the selection of convalescent plasma donors. Virus neutralisation tests (NTs) are the gold standard for the detection and quantification of NAbs, but they are complex and require BSL3 facilities. In contrast, surrogate enzyme-linked immunosorbent assays (sELISA) offer the possibility of high-throughput testing under standard laboratory safety conditions. In this study, we investigated two commercial sELISA kits (GenScript, AdipoGen) designed for the detection of SARS-CoV-2 NAbs. 276 plasma samples were screened using commercial IgG-ELISA and NAbs titres were determined by micro-neutralisation test (micro-NT). In addition, all samples were tested in both sELISA. Sensitivity and specificity for both sELISA were determined in comparison to the micro-NT results. 57 % of the samples were SARS-CoV-2 NAb positive in micro-NT, while 43 % tested negative. Comparison with micro-NT results showed a sensitivity of 98.2 % and a specificity of 69.5 % for the GenScript ELISA. The AdipoGen ELISA had a sensitivity of 83.5 % and a specificity of 97.8 %. False negative results were obtained mainly on samples with low NAbs titres. Both sELISA were able to qualitatively detect NAbs in plasma samples. Sensitivity and specificity differed between sELISA with GenScript superior in sensitivity and AdipoGen superior in specificity. Both sELISA were unable to quantify NAbs, thus neither of them can completely replace conventional NTs. However, in a two-step diagnostic algorithm, AdipoGen could potentially replace NT as a subsequent confirmatory test due to its high specificity but only in settings where no exact NAbs quantification is needed.

ACS Style

Katharina Müller; Philipp Girl; Heiner von Buttlar; Gerhard Dobler; Roman Wölfel. Comparison of two commercial surrogate ELISAs to detect a neutralising antibody response to SARS-CoV-2. Journal of Virological Methods 2021, 292, 114122 -114122.

AMA Style

Katharina Müller, Philipp Girl, Heiner von Buttlar, Gerhard Dobler, Roman Wölfel. Comparison of two commercial surrogate ELISAs to detect a neutralising antibody response to SARS-CoV-2. Journal of Virological Methods. 2021; 292 ():114122-114122.

Chicago/Turabian Style

Katharina Müller; Philipp Girl; Heiner von Buttlar; Gerhard Dobler; Roman Wölfel. 2021. "Comparison of two commercial surrogate ELISAs to detect a neutralising antibody response to SARS-CoV-2." Journal of Virological Methods 292, no. : 114122-114122.

Journal article
Published: 02 November 2020 in Viruses
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We are currently facing a pandemic of COVID-19, caused by a spillover from an animal-originating coronavirus to humans occurring in the Wuhan region of China in December 2019. From China, the virus has spread to 188 countries and regions worldwide, reaching the Sahel region on March 2, 2020. Since whole genome sequencing (WGS) data is very crucial to understand the spreading dynamics of the ongoing pandemic, but only limited sequencing data is available from the Sahel region to date, we have focused our efforts on generating the first Malian sequencing data available. Screening 217 Malian patient samples for the presence of SARS-CoV-2 resulted in 38 positive isolates, from which 21 whole genome sequences were generated. Our analysis shows that both the early A (19B) and the later observed B (20A/C) clade are present in Mali, indicating multiple and independent introductions of SARS-CoV-2 to the Sahel region.

ACS Style

Bourema Kouriba; Angela Dürr; Alexandra Rehn; Abdoul Sangaré; Brehima Traoré; Malena Bestehorn-Willmann; Judicael Ouedraogo; Asli Heitzer; Elisabeth Sogodogo; Abderrhamane Maiga; Mathias Walter; Fee Zimmermann; Roman Wölfel; Markus Antwerpen. First Phylogenetic Analysis of Malian SARS-CoV-2 Sequences Provides Molecular Insights into the Genomic Diversity of the Sahel Region. Viruses 2020, 12, 1251 .

AMA Style

Bourema Kouriba, Angela Dürr, Alexandra Rehn, Abdoul Sangaré, Brehima Traoré, Malena Bestehorn-Willmann, Judicael Ouedraogo, Asli Heitzer, Elisabeth Sogodogo, Abderrhamane Maiga, Mathias Walter, Fee Zimmermann, Roman Wölfel, Markus Antwerpen. First Phylogenetic Analysis of Malian SARS-CoV-2 Sequences Provides Molecular Insights into the Genomic Diversity of the Sahel Region. Viruses. 2020; 12 (11):1251.

Chicago/Turabian Style

Bourema Kouriba; Angela Dürr; Alexandra Rehn; Abdoul Sangaré; Brehima Traoré; Malena Bestehorn-Willmann; Judicael Ouedraogo; Asli Heitzer; Elisabeth Sogodogo; Abderrhamane Maiga; Mathias Walter; Fee Zimmermann; Roman Wölfel; Markus Antwerpen. 2020. "First Phylogenetic Analysis of Malian SARS-CoV-2 Sequences Provides Molecular Insights into the Genomic Diversity of the Sahel Region." Viruses 12, no. 11: 1251.

Other
Published: 31 July 2020
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In the current pandemic of SARS-CoV-2, rapid identification of infected individuals is crucial for management and control of the outbreak. However, transport of samples, sample processing and RT-qPCR analysis in laboratories are time-consuming. Here we present a nucleic acid-based test format – pulse controlled amplification – that allows detection of SARS-CoV-2 directly from up to eight swab samples simultaneously without the need for RNA extraction within 20 min in a point-of-care setting.

ACS Style

Zwirglmaier Katrin; Weyh Maria; Krüger Christian; Ehmann Rosina; Müller Katharina; Wölfel Roman; Stoecker Kilian. Rapid detection of SARS-CoV-2 by pulse-controlled amplification (PCA). 2020, 1 .

AMA Style

Zwirglmaier Katrin, Weyh Maria, Krüger Christian, Ehmann Rosina, Müller Katharina, Wölfel Roman, Stoecker Kilian. Rapid detection of SARS-CoV-2 by pulse-controlled amplification (PCA). . 2020; ():1.

Chicago/Turabian Style

Zwirglmaier Katrin; Weyh Maria; Krüger Christian; Ehmann Rosina; Müller Katharina; Wölfel Roman; Stoecker Kilian. 2020. "Rapid detection of SARS-CoV-2 by pulse-controlled amplification (PCA)." , no. : 1.

Journal article
Published: 20 May 2020 in Microorganisms
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A variety of methods have been established in order to optimize the accessibility of DNA originating from Bacillus anthracis cells and endospores to facilitate highly sensitive molecular diagnostics. However, most endospore lysis techniques have not been evaluated in respect to their quantitative proficiencies. Here, we started by systematically assessing the efficiencies of 20 DNA extraction kits for vegetative B. anthracis cells. Of these, the Epicentre MasterPure kit gave the best DNA yields and quality suitable for further genomic analysis. Yet, none of the kits tested were able to extract reasonable quantities of DNA from cores of the endospores. Thus, we developed a mechanical endospore lysis protocol, facilitating the extraction of high-quality DNA. Transmission electron microscopy or the labelling of spores with the indicator dye propidium monoazide was utilized to assess lysis efficiency. Finally, the yield and quality of genomic spore DNA were quantified by PCR and they were found to be dependent on lysis matrix composition, instrumental parameters, and the method used for subsequent DNA purification. Our final standardized lysis and DNA extraction protocol allows for the quantitative detection of low levels (

ACS Style

Mandy Knüpfer; Peter Braun; Kathrin Baumann; Alexandra Rehn; Markus Antwerpen; Gregor Grass; And Roman Wölfel. Evaluation of a Highly Efficient DNA Extraction Method for Bacillus anthracis Endospores. Microorganisms 2020, 8, 763 .

AMA Style

Mandy Knüpfer, Peter Braun, Kathrin Baumann, Alexandra Rehn, Markus Antwerpen, Gregor Grass, And Roman Wölfel. Evaluation of a Highly Efficient DNA Extraction Method for Bacillus anthracis Endospores. Microorganisms. 2020; 8 (5):763.

Chicago/Turabian Style

Mandy Knüpfer; Peter Braun; Kathrin Baumann; Alexandra Rehn; Markus Antwerpen; Gregor Grass; And Roman Wölfel. 2020. "Evaluation of a Highly Efficient DNA Extraction Method for Bacillus anthracis Endospores." Microorganisms 8, no. 5: 763.

Other
Published: 08 March 2020
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Coronavirus disease 2019 (COVID-19) is an acute respiratory tract infection that emerged in late 20191,2. Initial outbreaks in China involved 13.8% cases with severe-, and 6.1% with critical courses3. This severe presentation corresponds to the usage of a virus receptor that is expressed predominantly in the lung2,4. By causing an early onset of severe symptoms, this same receptor tropism is thought to have determined pathogenicity but also aided the control of severe acute respiratory syndrome (SARS) in 20035. However, there are reports of COVID-19 cases with mild upper respiratory tract symptoms, suggesting a potential for pre- or oligosymptomatic transmission6-8. There is an urgent need for information on body site - specific virus replication, immunity, and infectivity. Here we provide a detailed virological analysis of nine cases, providing proof of active virus replication in upper respiratory tract tissues. Pharyngeal virus shedding was very high during the first week of symptoms (peak at 7.11 × 108 RNA copies per throat swab, day 4). Infectious virus was readily isolated from throat- and lung-derived samples, but not from stool samples in spite of high virus RNA concentration. Blood and urine never yielded virus. Active replication in the throat was confirmed by viral replicative RNA intermediates in throat samples. Sequence-distinct virus populations were consistently detected in throat- and lung samples of one same patient. Shedding of viral RNA from sputum outlasted the end of symptoms. Seroconversion occurred after 6-12 days, but was not followed by a rapid decline of viral loads. COVID-19 can present as a mild upper respiratory tract illness. Active virus replication in the upper respiratory tract puts prospects of COVID-19 containment in perspective.

ACS Style

Roman Woelfel; Victor Max Corman; Wolfgang Guggemos; Michael Seilmaier; Sabine Zange; Marcel A. Müller; Daniela Niemeyer; Terence C. Jones Kelly; Patrick Vollmar; Camilla Rothe; Michael Hoelscher; Tobias Bleicker; Sebastian Bruenink; Julia Schneider; Rosina Ehmann; Katrin Zwirglmaier; Christian Drosten; Clemens Wendtner. Virological assessment of hospitalized cases of coronavirus disease 2019. 2020, 1 .

AMA Style

Roman Woelfel, Victor Max Corman, Wolfgang Guggemos, Michael Seilmaier, Sabine Zange, Marcel A. Müller, Daniela Niemeyer, Terence C. Jones Kelly, Patrick Vollmar, Camilla Rothe, Michael Hoelscher, Tobias Bleicker, Sebastian Bruenink, Julia Schneider, Rosina Ehmann, Katrin Zwirglmaier, Christian Drosten, Clemens Wendtner. Virological assessment of hospitalized cases of coronavirus disease 2019. . 2020; ():1.

Chicago/Turabian Style

Roman Woelfel; Victor Max Corman; Wolfgang Guggemos; Michael Seilmaier; Sabine Zange; Marcel A. Müller; Daniela Niemeyer; Terence C. Jones Kelly; Patrick Vollmar; Camilla Rothe; Michael Hoelscher; Tobias Bleicker; Sebastian Bruenink; Julia Schneider; Rosina Ehmann; Katrin Zwirglmaier; Christian Drosten; Clemens Wendtner. 2020. "Virological assessment of hospitalized cases of coronavirus disease 2019." , no. : 1.

Journal article
Published: 12 December 2019 in Microorganisms
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The largest phylogenetic lineage known to date of the anthrax pathogen Bacillus anthracis is the wide-spread, so-called Trans-Eurasian clade systematically categorized as the A.Br.008/009 group sharing two defining canonical single-nucleotide polymorphisms (canSNP). In this study, we genome-sequenced a collection of 35 B. anthracis strains of this clade, derived from human infections, animal outbreaks or soil, mostly from European countries isolated between 1936 and 2008. The new data were subjected to comparative chromosomal analysis, together with 75 B. anthracis genomes available in public databases, and the relative placements of these isolates were determined within the global phylogeny of the A.Br.008/009 canSNP group. From this analysis, we have detected 3754 chromosomal SNPs, allowing the assignation of the new chromosomal sequences to established sub-clades, to define new sub-clades, such as two new Spanish, one Bulgarian or one German group(s), or to introduce orphan lineages. SNP-based results were compared with that of a multilocus variable number of tandem repeat analysis (MLVA). This analysis indicated that MLVA typing might provide additional information in cases when genomics yields identical genotypes or shows only minor differences. Introducing the delayed mismatch amplification assay (DMAA) PCR-analysis, we developed a cost-effective method to interrogate for a set of ten phylogenetically informative SNPs within genomes of A.Br.008/009 canSNP clade strains of B. anthracis. By this approach, additional 32 strains could be assigned to five of ten defined clades.

ACS Style

Markus Antwerpen; Wolfgang Beyer; Olga Bassy; María Victoria Ortega-García; Juan Carlos Cabria-Ramos; Gregor Grass; Roman Wölfel. Phylogenetic Placement of Isolates Within the Trans-Eurasian Clade A.Br.008/009 of Bacillus anthracis. Microorganisms 2019, 7, 689 .

AMA Style

Markus Antwerpen, Wolfgang Beyer, Olga Bassy, María Victoria Ortega-García, Juan Carlos Cabria-Ramos, Gregor Grass, Roman Wölfel. Phylogenetic Placement of Isolates Within the Trans-Eurasian Clade A.Br.008/009 of Bacillus anthracis. Microorganisms. 2019; 7 (12):689.

Chicago/Turabian Style

Markus Antwerpen; Wolfgang Beyer; Olga Bassy; María Victoria Ortega-García; Juan Carlos Cabria-Ramos; Gregor Grass; Roman Wölfel. 2019. "Phylogenetic Placement of Isolates Within the Trans-Eurasian Clade A.Br.008/009 of Bacillus anthracis." Microorganisms 7, no. 12: 689.

Journal article
Published: 01 January 2017 in The Lancet Global Health
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Summary Background By January, 2016, all known transmission chains of the Ebola virus disease (EVD) outbreak in west Africa had been stopped. However, there is concern about persistence of Ebola virus in the reproductive tract of men who have survived EVD. We aimed to use biostatistical modelling to describe the dynamics of Ebola virus RNA load in seminal fluid, including clearance parameters. Methods In this longitudinal study, we recruited men who had been discharged from three Ebola treatment units in Guinea between January and July, 2015. Participants provided samples of seminal fluid at follow-up every 3–6 weeks, which we tested for Ebola virus RNA using quantitative real-time RT-PCR. Representative specimens from eight participants were then inoculated into immunodeficient mice to test for infectivity. We used a linear mixed-effect model to analyse the dynamics of virus persistence in seminal fluid over time. Findings We enrolled 26 participants and tested 130 seminal fluid specimens; median follow up was 197 days (IQR 187–209 days) after enrolment, which corresponded to 255 days (228–287) after disease onset. Ebola virus RNA was detected in 86 semen specimens from 19 (73%) participants. Median duration of Ebola virus RNA detection was 158 days after onset (73–181; maximum 407 days at end of follow-up). Mathematical modelling of the quantitative time-series data showed a mean clearance rate of Ebola virus RNA from seminal fluid of −0·58 log units per month, although the clearance kinetic varied greatly between participants. Using our biostatistical model, we predict that 50% and 90% of male survivors clear Ebola virus RNA from seminal fluid at 115 days (90% prediction interval 72–160) and 294 days (212–399) after disease onset, respectively. We also predicted that the number of men positive for Ebola virus RNA in affected countries would decrease from about 50 in January 2016, to fewer than 1 person by July, 2016. Infectious virus was detected in 15 of 26 (58%) specimens tested in mice. Interpretation Time to clearance of Ebola virus RNA from seminal fluid varies greatly between individuals and could be more than 13 months. Our predictions will assist in decision-making about surveillance and preventive measures in EVD outbreaks. Funding This study was funded by European Union's Horizon 2020 research and innovation programme, Directorate-General for International Cooperation and Development of the European Commission, Institut national de la santé et de la recherche médicale (INSERM), German Research Foundation (DFG), and Innovative Medicines Initiative 2 Joint Undertaking.

ACS Style

Daouda Sissoko; Sophie Duraffour; Romy Kerber; Jacques Seraphin Kolie; Abdoul Habib Beavogui; Alseny-Modet Camara; Géraldine Colin; Toni Rieger; Lisa Oestereich; Bernadett Pályi; Stephanie Wurr; Jeremie Guedj; Thi Huyen Tram Nguyen; Rosalind M Eggo; Conall Watson; W John Edmunds; Joseph Akoi Bore; Fara Raymond Koundouno; Mar Cabeza-Cabrerizo; Lisa L Carter; Liana Eleni Kafetzopoulou; Eeva Kuisma; Janine Michel; Livia Victoria Patrono; Natasha Rickett; Katrin Singethan; Martin Rudolf; Angelika Lander; Elisa Pallasch; Sabrina Bockholt; Estefanía Rodríguez; Antonino Di Caro; Roman Wölfel; Martin Gabriel; Céline Gurry; Pierre Formenty; Sakoba Keïta; Denis Malvy; Miles W Carroll; Xavier Anglaret; Stephan Günther. Persistence and clearance of Ebola virus RNA from seminal fluid of Ebola virus disease survivors: a longitudinal analysis and modelling study. The Lancet Global Health 2017, 5, e80 -e88.

AMA Style

Daouda Sissoko, Sophie Duraffour, Romy Kerber, Jacques Seraphin Kolie, Abdoul Habib Beavogui, Alseny-Modet Camara, Géraldine Colin, Toni Rieger, Lisa Oestereich, Bernadett Pályi, Stephanie Wurr, Jeremie Guedj, Thi Huyen Tram Nguyen, Rosalind M Eggo, Conall Watson, W John Edmunds, Joseph Akoi Bore, Fara Raymond Koundouno, Mar Cabeza-Cabrerizo, Lisa L Carter, Liana Eleni Kafetzopoulou, Eeva Kuisma, Janine Michel, Livia Victoria Patrono, Natasha Rickett, Katrin Singethan, Martin Rudolf, Angelika Lander, Elisa Pallasch, Sabrina Bockholt, Estefanía Rodríguez, Antonino Di Caro, Roman Wölfel, Martin Gabriel, Céline Gurry, Pierre Formenty, Sakoba Keïta, Denis Malvy, Miles W Carroll, Xavier Anglaret, Stephan Günther. Persistence and clearance of Ebola virus RNA from seminal fluid of Ebola virus disease survivors: a longitudinal analysis and modelling study. The Lancet Global Health. 2017; 5 (1):e80-e88.

Chicago/Turabian Style

Daouda Sissoko; Sophie Duraffour; Romy Kerber; Jacques Seraphin Kolie; Abdoul Habib Beavogui; Alseny-Modet Camara; Géraldine Colin; Toni Rieger; Lisa Oestereich; Bernadett Pályi; Stephanie Wurr; Jeremie Guedj; Thi Huyen Tram Nguyen; Rosalind M Eggo; Conall Watson; W John Edmunds; Joseph Akoi Bore; Fara Raymond Koundouno; Mar Cabeza-Cabrerizo; Lisa L Carter; Liana Eleni Kafetzopoulou; Eeva Kuisma; Janine Michel; Livia Victoria Patrono; Natasha Rickett; Katrin Singethan; Martin Rudolf; Angelika Lander; Elisa Pallasch; Sabrina Bockholt; Estefanía Rodríguez; Antonino Di Caro; Roman Wölfel; Martin Gabriel; Céline Gurry; Pierre Formenty; Sakoba Keïta; Denis Malvy; Miles W Carroll; Xavier Anglaret; Stephan Günther. 2017. "Persistence and clearance of Ebola virus RNA from seminal fluid of Ebola virus disease survivors: a longitudinal analysis and modelling study." The Lancet Global Health 5, no. 1: e80-e88.

Journal article
Published: 01 January 2017 in Ticks and Tick-borne Diseases
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In the last decade six Rickettsia species, including Rickettsia slovaca have been characterized in Germany. All of these species could be linked to distinct clinical syndromes in humans. However, due to lack of seroepidemiological data an estimation of the prevalence and the public health impact of rickettsial infections in Germany is difficult. The aim of the present study was to determine the seroprevalence of spotted fever group (SFG) rickettsiae in a population with an elevated exposure risk to ticks. For that purpose, 559 sera of forestry workers in the federal state of Brandenburg, Eastern Germany, were screened for SFG-rickettsiae reactive IgG antibodies. Positive sera were subsequently titrated by microimmunofluorescence assay against R. helvetica, R. raoultii, R. felis, "R. monacensis" and R. slovaca. The total average IgG seroprevalence rate against SFG rickettsiae of 27.5% was found to be represented by 9.7% R. helvetica, 5% R. raoultii, 2.7% R. felis, 0.5% "R. monacensis" and 0.5% R. slovaca. The remaining 9.1% positive test results were of non-differentiable origin. IgG seroprevalences ranged from 11% to 55% in the different forestry districts. Older and male participants had a significantly higher probability for seropositivity and higher anti-rickettsia antibody titer level. In addition, the number of recent as well as the recalled lifetime tick bites was significantly associated with seropositivity and higher titers against SFG rickettsiae. In conclusion, we found an unexpected high total seroprevalence against SFG rickettsiae in forestry workers and serological evidence confirming the occurrence of R. raoultii, R. felis, "R. monacensis" and R. helvetica in the federal State of Brandenburg.

ACS Style

Silke Wölfel; Stephanie Speck; Sandra Essbauer; Bryan R. Thoma; Marc Mertens; Sandra Werdermann; Olaf Niederstrasser; Eckhardt Petri; Rainer G. Ulrich; Roman Wölfel; Gerhard Dobler. High seroprevalence for indigenous spotted fever group rickettsiae in forestry workers from the federal state of Brandenburg, Eastern Germany. Ticks and Tick-borne Diseases 2017, 8, 132 -138.

AMA Style

Silke Wölfel, Stephanie Speck, Sandra Essbauer, Bryan R. Thoma, Marc Mertens, Sandra Werdermann, Olaf Niederstrasser, Eckhardt Petri, Rainer G. Ulrich, Roman Wölfel, Gerhard Dobler. High seroprevalence for indigenous spotted fever group rickettsiae in forestry workers from the federal state of Brandenburg, Eastern Germany. Ticks and Tick-borne Diseases. 2017; 8 (1):132-138.

Chicago/Turabian Style

Silke Wölfel; Stephanie Speck; Sandra Essbauer; Bryan R. Thoma; Marc Mertens; Sandra Werdermann; Olaf Niederstrasser; Eckhardt Petri; Rainer G. Ulrich; Roman Wölfel; Gerhard Dobler. 2017. "High seroprevalence for indigenous spotted fever group rickettsiae in forestry workers from the federal state of Brandenburg, Eastern Germany." Ticks and Tick-borne Diseases 8, no. 1: 132-138.

Preprint
Published: 02 September 2016
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SummaryThe 2013-2016 epidemic of Ebola virus disease in West Africa was of unprecedented magnitude, duration and impact. Extensive collaborative sequencing projects have produced a large collection of over 1600 Ebola virus genomes, representing over 5% of known cases, unmatched for any single human epidemic. In this comprehensive analysis of this entire dataset, we reconstruct in detail the history of migration, proliferation and decline of Ebola virus throughout the region. We test the association of geography, climate, administrative boundaries, demography and culture with viral movement among 56 administrative regions. Our results show that during the outbreak viral lineages moved according to a classic ‘gravity’ model, with more intense migration between larger and more proximate population centers. Notably, we find that despite a strong attenuation of international dispersal after border closures, localized cross-border transmission beforehand had already set the seeds for an international epidemic, rendering these measures relatively ineffective in curbing the epidemic. We use this empirical evidence to address why the epidemic did not spread into neighboring countries, showing that although these regions were susceptible to developing significant outbreaks, they were also at lower risk of viral introductions. Finally, viral genome sequence data uniquely reveals this large epidemic to be a heterogeneous and spatially dissociated collection of transmission clusters of varying size, duration and connectivity. These insights will help inform approaches to intervention in such epidemics in the future.

ACS Style

Gytis Dudas; Luiz Max Carvalho; Trevor Bedford; Andrew J. Tatem; Guy Baele; Nuno Faria; Daniel J. Park; Jason Ladner; Armando Arias; Danny Asogun; Filip Bielejec; Sarah Caddy; Matt Cotten; Jonathan Dambrozio; Simon Dellicour; Antonino Di Caro; Joseph W. Diclaro; Sophie Duraffour; Mike Elmore; Lawrence Fakoli; Merle Gilbert; Sahr M. Gevao; Stephen Gire; Adrianne Gladden-Young; Andreas Gnirke; Augustine Goba; Donald S. Grant; Bart Haagmans; Julian A. Hiscox; Umaru Jah; Brima Kargbo; Jeffrey Kugelman; Di Liu; Jia Lu; Christine M. Malboeuf; Suzanne Mate; David A. Matthews; Christian B. Matranga; Luke Meredith; James Qu; Joshua Quick; Susan D. Pas; My Vt Phan; Georgios Poliakis; Chantal Reusken; Mariano Sanchez-Lockhart; Stephen F. Schaffner; John S. Schieffelin; Rachel S. Sealfon; Etienne Simon-Loriere; Saskia L. Smits; Kilian Stoecker; Lucy Thorne; Ekaete Alice Tobin; Mohamed A. Vandi; Simon J. Watson; Kendra West; Shannon Whitmer; Michael R. Wiley; Sarah M. Winnicki; Shirlee Wohl; Roman Wölfel; Nathan L. Yozwiak; Kristian G. Andersen; Sylvia Blyden; Fatorma Bolay; Miles Carroll; Bernice Dahn; Boubacar Diallo; Pierre Formenty; Christophe Fraser; George F. Gao; Robert F. Garry; Ian Goodfellow; Stephan Günther; Christian Happi; Edward C Holmes; Paul Kellam; Marion P.G. Koopmans; Nicholas J. Loman; N'faly Magassouba; Dhamari Naidoo; Stuart T. Nichol; Tolbert Nyenswah; Gustavo Palacios; Oliver G Pybus; Pardis Sabeti; Amadou Sall; Keïta Sakoba; Ute Ströeher; Isatta Wurie; Marc A Suchard; Philippe Lemey; Andrew Rambaut; Marc Suchard. Virus genomes reveal the factors that spread and sustained the West African Ebola epidemic. 2016, 071779 .

AMA Style

Gytis Dudas, Luiz Max Carvalho, Trevor Bedford, Andrew J. Tatem, Guy Baele, Nuno Faria, Daniel J. Park, Jason Ladner, Armando Arias, Danny Asogun, Filip Bielejec, Sarah Caddy, Matt Cotten, Jonathan Dambrozio, Simon Dellicour, Antonino Di Caro, Joseph W. Diclaro, Sophie Duraffour, Mike Elmore, Lawrence Fakoli, Merle Gilbert, Sahr M. Gevao, Stephen Gire, Adrianne Gladden-Young, Andreas Gnirke, Augustine Goba, Donald S. Grant, Bart Haagmans, Julian A. Hiscox, Umaru Jah, Brima Kargbo, Jeffrey Kugelman, Di Liu, Jia Lu, Christine M. Malboeuf, Suzanne Mate, David A. Matthews, Christian B. Matranga, Luke Meredith, James Qu, Joshua Quick, Susan D. Pas, My Vt Phan, Georgios Poliakis, Chantal Reusken, Mariano Sanchez-Lockhart, Stephen F. Schaffner, John S. Schieffelin, Rachel S. Sealfon, Etienne Simon-Loriere, Saskia L. Smits, Kilian Stoecker, Lucy Thorne, Ekaete Alice Tobin, Mohamed A. Vandi, Simon J. Watson, Kendra West, Shannon Whitmer, Michael R. Wiley, Sarah M. Winnicki, Shirlee Wohl, Roman Wölfel, Nathan L. Yozwiak, Kristian G. Andersen, Sylvia Blyden, Fatorma Bolay, Miles Carroll, Bernice Dahn, Boubacar Diallo, Pierre Formenty, Christophe Fraser, George F. Gao, Robert F. Garry, Ian Goodfellow, Stephan Günther, Christian Happi, Edward C Holmes, Paul Kellam, Marion P.G. Koopmans, Nicholas J. Loman, N'faly Magassouba, Dhamari Naidoo, Stuart T. Nichol, Tolbert Nyenswah, Gustavo Palacios, Oliver G Pybus, Pardis Sabeti, Amadou Sall, Keïta Sakoba, Ute Ströeher, Isatta Wurie, Marc A Suchard, Philippe Lemey, Andrew Rambaut, Marc Suchard. Virus genomes reveal the factors that spread and sustained the West African Ebola epidemic. . 2016; ():071779.

Chicago/Turabian Style

Gytis Dudas; Luiz Max Carvalho; Trevor Bedford; Andrew J. Tatem; Guy Baele; Nuno Faria; Daniel J. Park; Jason Ladner; Armando Arias; Danny Asogun; Filip Bielejec; Sarah Caddy; Matt Cotten; Jonathan Dambrozio; Simon Dellicour; Antonino Di Caro; Joseph W. Diclaro; Sophie Duraffour; Mike Elmore; Lawrence Fakoli; Merle Gilbert; Sahr M. Gevao; Stephen Gire; Adrianne Gladden-Young; Andreas Gnirke; Augustine Goba; Donald S. Grant; Bart Haagmans; Julian A. Hiscox; Umaru Jah; Brima Kargbo; Jeffrey Kugelman; Di Liu; Jia Lu; Christine M. Malboeuf; Suzanne Mate; David A. Matthews; Christian B. Matranga; Luke Meredith; James Qu; Joshua Quick; Susan D. Pas; My Vt Phan; Georgios Poliakis; Chantal Reusken; Mariano Sanchez-Lockhart; Stephen F. Schaffner; John S. Schieffelin; Rachel S. Sealfon; Etienne Simon-Loriere; Saskia L. Smits; Kilian Stoecker; Lucy Thorne; Ekaete Alice Tobin; Mohamed A. Vandi; Simon J. Watson; Kendra West; Shannon Whitmer; Michael R. Wiley; Sarah M. Winnicki; Shirlee Wohl; Roman Wölfel; Nathan L. Yozwiak; Kristian G. Andersen; Sylvia Blyden; Fatorma Bolay; Miles Carroll; Bernice Dahn; Boubacar Diallo; Pierre Formenty; Christophe Fraser; George F. Gao; Robert F. Garry; Ian Goodfellow; Stephan Günther; Christian Happi; Edward C Holmes; Paul Kellam; Marion P.G. Koopmans; Nicholas J. Loman; N'faly Magassouba; Dhamari Naidoo; Stuart T. Nichol; Tolbert Nyenswah; Gustavo Palacios; Oliver G Pybus; Pardis Sabeti; Amadou Sall; Keïta Sakoba; Ute Ströeher; Isatta Wurie; Marc A Suchard; Philippe Lemey; Andrew Rambaut; Marc Suchard. 2016. "Virus genomes reveal the factors that spread and sustained the West African Ebola epidemic." , no. : 071779.

Journal article
Published: 28 July 2015 in The Lancet Infectious Diseases
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ACS Style

Giuseppe Ippolito; Simone Lanini; Philippe Brouqui; Antonino Di Caro; Francesco Vairo; Salim Abdulla; Francesco Maria Fusco; Sanjeev Krishna; Maria Rosaria Capobianchi; Henry Kyobe Bosa; David J M Lewis; Vincenzo Puro; Roman Wolfel; Tatjana Avšič Županc; Osman Dar; Peter Mwaba; Matthew Bates; David Heymann; Professor Sir Alimuddin Zumla. Ebola: missed opportunities for Europe–Africa research. The Lancet Infectious Diseases 2015, 15, 1254 -1255.

AMA Style

Giuseppe Ippolito, Simone Lanini, Philippe Brouqui, Antonino Di Caro, Francesco Vairo, Salim Abdulla, Francesco Maria Fusco, Sanjeev Krishna, Maria Rosaria Capobianchi, Henry Kyobe Bosa, David J M Lewis, Vincenzo Puro, Roman Wolfel, Tatjana Avšič Županc, Osman Dar, Peter Mwaba, Matthew Bates, David Heymann, Professor Sir Alimuddin Zumla. Ebola: missed opportunities for Europe–Africa research. The Lancet Infectious Diseases. 2015; 15 (11):1254-1255.

Chicago/Turabian Style

Giuseppe Ippolito; Simone Lanini; Philippe Brouqui; Antonino Di Caro; Francesco Vairo; Salim Abdulla; Francesco Maria Fusco; Sanjeev Krishna; Maria Rosaria Capobianchi; Henry Kyobe Bosa; David J M Lewis; Vincenzo Puro; Roman Wolfel; Tatjana Avšič Županc; Osman Dar; Peter Mwaba; Matthew Bates; David Heymann; Professor Sir Alimuddin Zumla. 2015. "Ebola: missed opportunities for Europe–Africa research." The Lancet Infectious Diseases 15, no. 11: 1254-1255.

Journal article
Published: 31 August 2012 in Toxins
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Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR) assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix.

ACS Style

Eva Felder; Ilona Mossbrugger; Mirko Lange; Roman Wölfel. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR). Toxins 2012, 4, 633 -642.

AMA Style

Eva Felder, Ilona Mossbrugger, Mirko Lange, Roman Wölfel. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR). Toxins. 2012; 4 (9):633-642.

Chicago/Turabian Style

Eva Felder; Ilona Mossbrugger; Mirko Lange; Roman Wölfel. 2012. "Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR)." Toxins 4, no. 9: 633-642.

Evaluation study
Published: 01 April 2009 in Journal of Clinical Microbiology
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Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral zoonosis which occurs throughout Africa, Eastern Europe, and Asia and results in an approximately 30% fatality rate. A reverse transcription-PCR assay including a competitive internal control was developed on the basis of the most up-to-date genome information. Biotinylated amplification products were hybridized to DNA macroarrays on the surfaces of polymer supports, and hybridization events were visualized by incubation with a streptavidin-horseradish peroxidase conjugate and the formation of a visible substrate precipitate. Optimal assay conditions for the detection of as few as 6.3 genome copies per reaction were established. Eighteen geographically and historically diverse CCHF virus strains representing all clinically relevant isolates were detected. The feasibility of the assay for clinical diagnosis was validated with acute-phase patient samples from South Africa, Iran, and Pakistan. The assay provides a specific, sensitive, and rapid method for CCHF virus detection without requiring sophisticated equipment. It has usefulness for the clinical diagnosis and surveillance of CCHF infections under limited laboratory conditions in developing countries or in field situations.

ACS Style

Roman Wölfel; Janusz T. Paweska; Nadine Petersen; Antoinette A. Grobbelaar; Patricia A. Leman; Roger Hewson; Marie-Claude Georges-Courbot; Anna Papa; Volker Heiser; Marcus Panning; Stephan Günther; Christian Drosten. Low-Density Macroarray for Rapid Detection and Identification of Crimean-Congo Hemorrhagic Fever Virus. Journal of Clinical Microbiology 2009, 47, 1025 -1030.

AMA Style

Roman Wölfel, Janusz T. Paweska, Nadine Petersen, Antoinette A. Grobbelaar, Patricia A. Leman, Roger Hewson, Marie-Claude Georges-Courbot, Anna Papa, Volker Heiser, Marcus Panning, Stephan Günther, Christian Drosten. Low-Density Macroarray for Rapid Detection and Identification of Crimean-Congo Hemorrhagic Fever Virus. Journal of Clinical Microbiology. 2009; 47 (4):1025-1030.

Chicago/Turabian Style

Roman Wölfel; Janusz T. Paweska; Nadine Petersen; Antoinette A. Grobbelaar; Patricia A. Leman; Roger Hewson; Marie-Claude Georges-Courbot; Anna Papa; Volker Heiser; Marcus Panning; Stephan Günther; Christian Drosten. 2009. "Low-Density Macroarray for Rapid Detection and Identification of Crimean-Congo Hemorrhagic Fever Virus." Journal of Clinical Microbiology 47, no. 4: 1025-1030.

Journal article
Published: 01 September 2008 in International Journal of Medical Microbiology
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Tick-borne rickettsioses in humans occur worldwide and are caused by obligate intracellular bacteria belonging to the spotted-fever group (SFG) within the genus Rickettsia (R.). These tick-borne rickettsioses are among the most underdiagnosed vector-borne diseases in Germany: Due to the variety of unspecific clinical signs, they are not easily recognised. The clinical picture ranges from subclinical to fatal courses, but may be difficult to differentiate from other febrile conditions without specific tests. Even to date, diagnosis is either based on clinical findings, a record of tick exposure, or indirect serological detection of the pathogens. Herein, we briefly discuss modern diagnostic tools for important tick-borne rickettsial infections with emphasis on new molecular diagnostic assays. As one example, we present a novel real-time polymerase chain reaction (PCR) protocol that facilitates genus-specific, rapid, and sensitive detection of rickettsial pathogens. A conserved region of the rickettsial citrate synthase gene (gltA) is amplified and detected by a 5′-nuclease probe in a LightCycler instrument. Sensitivity was consistently high at less than 23 genome copies per reaction. This detection system has been evaluated both as a useful tool in epidemiological investigations in ticks and in human diagnostics. We describe a rational diagnostic approach for the detection of tick-borne human rickettsioses which consists of that real-time PCR, isolation of rickettsiae in cell culture, multi-locus sequence typing, and serology. Its implementation recently led to the first isolation and characterisation of R. africae in Germany from a patient returning from Zimbabwe. In conclusion, tick-borne rickettsioses should be considered in patients presenting with fever, headache, and rash following a tick bite. Further studies are needed to determine the epidemiology and clinical importance of these pathogens in Germany.

ACS Style

Roman Wölfel; Sandra Essbauer; Gerhard Dobler. Diagnostics of tick-borne rickettsioses in Germany: A modern concept for a neglected disease. International Journal of Medical Microbiology 2008, 298, 368 -374.

AMA Style

Roman Wölfel, Sandra Essbauer, Gerhard Dobler. Diagnostics of tick-borne rickettsioses in Germany: A modern concept for a neglected disease. International Journal of Medical Microbiology. 2008; 298 ():368-374.

Chicago/Turabian Style

Roman Wölfel; Sandra Essbauer; Gerhard Dobler. 2008. "Diagnostics of tick-borne rickettsioses in Germany: A modern concept for a neglected disease." International Journal of Medical Microbiology 298, no. : 368-374.

Comparative study
Published: 21 March 2006 in Graefe's Archive for Clinical and Experimental Ophthalmology
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Human adenoviruses (HAdV) may cause pharyngoconjunctival fever, follicular conjunctivitis or epidemic keratoconjunctivitis (EKC). Especially, outbreaks of the latter may lead to severe economic losses when preventive measures are implemented too late. Thus, a safe sampling method, proper specimen transport conditions and a fast and sensitive diagnostic technique is mandatory. Two commercially available virus transport systems (VTS) were compared with two NaCl-moisturised sampling devices, one of which comprises Dacron-tipped plastic-shafted swabs and the other a cotton-tipped wood-shafted swab, available in most ophthalmologists' offices. Downstream methods for specific detection of HAdV included direct immunofluorescence assay (IFA) of conjunctival swabs, virus isolation by cell culture and quantitative real-time polymerase chain reaction (qPCR). Furthermore, the influence of application of local anaesthetics prior to swabbing on subsequent detection of HAdV was investigated. Application of local anaesthetics had a positive influence on the amount of swabbed cells, thus increasing the chance of obtaining positive results by IFA. Neither isolation of HAdV by cell culture nor by qPCR was negatively influenced by this pretreatment. Surprisingly, both commercially available VTS performed significantly worse than the NaCl-moisturised swabs. This was shown with regard to virus recovery rates in cell culture as well as viral genome copy numbers in the qPCR. Based on our results, the following recommendations are provided to improve sampling, transport and diagnostic techniques regarding conjunctival swabs for diagnosis of human adenovirus infection: (1) application of local anaesthetics, (2) NaCl-moisturised VTS for shipment of specimens, and (3) detection of HAdV by qPCR. The latter method proved to be superior to virus isolation by cell culture, including subsequent identification by IFA, because it is faster, more sensitive and allows simultaneous handling of a number of samples. Hence, countermeasures to prevent further virus spread in an outbreak situation can be implemented earlier, thus reducing the number of subsequent adenoviral infections.

ACS Style

Roman Wölfel; Martin Pfeffer; Sandra Essbauer; Sylke Nerkelun; Gerhard Dobler. Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections. Graefe's Archive for Clinical and Experimental Ophthalmology 2006, 244, 1497 -1504.

AMA Style

Roman Wölfel, Martin Pfeffer, Sandra Essbauer, Sylke Nerkelun, Gerhard Dobler. Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections. Graefe's Archive for Clinical and Experimental Ophthalmology. 2006; 244 (11):1497-1504.

Chicago/Turabian Style

Roman Wölfel; Martin Pfeffer; Sandra Essbauer; Sylke Nerkelun; Gerhard Dobler. 2006. "Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections." Graefe's Archive for Clinical and Experimental Ophthalmology 244, no. 11: 1497-1504.