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Dr. Emma Bentley
National Institute for Biological Standards and Control-MHRA

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0 Molecular Virology
0 Serology
0 Virology
0 zoonosis
0 Public Health

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Journal article
Published: 25 August 2021 in Tropical Medicine and Infectious Disease
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Ebolaviruses continue to pose a significant outbreak threat, and while Ebola virus (EBOV)-specific vaccines and antivirals have been licensed, efforts to develop candidates offering broad species cross-protection are continuing. The use of pseudotyped virus in place of live virus is recognised as an alternative, safer, high-throughput platform to evaluate anti-ebolavirus antibodies towards their development, yet it requires optimisation. Here, we have shown that the target cell line impacts neutralisation assay results and cannot be selected purely based on permissiveness. In expanding the platform to incorporate each of the ebolavirus species envelope glycoprotein, allowing a comprehensive assessment of cross-neutralisation, we found that the recently discovered Bombali virus has a point mutation in the receptor-binding domain which prevents entry into a hamster cell line and, importantly, shows that this virus can be cross-neutralised by EBOV antibodies and convalescent plasma.

ACS Style

Emma M. Bentley; Samuel Richardson; Mariliza Derveni; Pramila Rijal; Alain R. Townsend; Jonathan L. Heeney; Giada Mattiuzzo; Edward Wright. Cross-Neutralisation of Novel Bombali Virus by Ebola Virus Antibodies and Convalescent Plasma Using an Optimised Pseudotype-Based Neutralisation Assay. Tropical Medicine and Infectious Disease 2021, 6, 155 .

AMA Style

Emma M. Bentley, Samuel Richardson, Mariliza Derveni, Pramila Rijal, Alain R. Townsend, Jonathan L. Heeney, Giada Mattiuzzo, Edward Wright. Cross-Neutralisation of Novel Bombali Virus by Ebola Virus Antibodies and Convalescent Plasma Using an Optimised Pseudotype-Based Neutralisation Assay. Tropical Medicine and Infectious Disease. 2021; 6 (3):155.

Chicago/Turabian Style

Emma M. Bentley; Samuel Richardson; Mariliza Derveni; Pramila Rijal; Alain R. Townsend; Jonathan L. Heeney; Giada Mattiuzzo; Edward Wright. 2021. "Cross-Neutralisation of Novel Bombali Virus by Ebola Virus Antibodies and Convalescent Plasma Using an Optimised Pseudotype-Based Neutralisation Assay." Tropical Medicine and Infectious Disease 6, no. 3: 155.

Article
Published: 26 May 2021
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Background The rise of SARS-CoV-2 variants has made the pursuit to define correlates of protection more troublesome, despite the availability of the World Health Organisation (WHO) International Standard for anti-SARS-CoV-2 Immunoglobulin sera, a key reagent used to standardise laboratory findings into an international unitage. Methods Using pseudotyped virus, we examine the capacity of convalescent sera, from a well-defined cohort of healthcare workers (HCW) and Patients infected during the first wave from a national critical care centre in the UK to neutralise B.1.1.298, variants of interest (VOI) B.1.617.1 (Kappa), and four VOCs, B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma) and B.1.617.2 (Delta), including the B.1.617.2 K417N, informally known as Delta Plus. We utilised the WHO International Standard for anti-SARS-CoV-2 Immunoglobulin to report neutralisation antibody levels in International Units per mL. Findings Our data demonstrate a significant reduction in the ability of first wave convalescent sera to neutralise the VOCs. Patients and HCWs with more severe COVID-19 were found to have higher antibody titres and to neutralise the VOCs more effectively than individuals with milder symptoms. Using an estimated threshold for 50% protection, 54 IU/mL, we found most asymptomatic and mild cases did not produce titres above this threshold. Interpretation Expressing our data in IU/ml, we provide a benchmark pre-vaccine standardised dataset that compares disease severity with neutralising antibody titres. Our data may now be compared across multiple laboratories. The continued use and aggregation of standardised data will eventually assist in defining correlates of protection. Funding UKRI and NIHR; grant number G107217 Research in context Evidence before this study During the first wave outbreak, much focus was placed on the role of neutralising antibodies and titres generated upon infection to ancestral SARS-CoV-2. Due to the large amounts of different assays used to elucidate the antibody-mediated immunity and laboratory to laboratory, large amounts of invaluable data could not be directly compared in order to define a correlate of protection, due to variability in the results. The WHO International Standard for anti-SARS-CoV-2 Immunoglobulin sera was made in order to standardise future data so that comparisons may take place. Added value of this study Our study compares the neutralisation capacity of sera from patients and healthcare workers (HCWs) from the ancestral strain of SARS-CoV-2 against new variants, including the current variants of concern in circulation. We also provide data in International Units per mL, a standardised unitage, for infected individuals that have a clinical severity score, allowing us to assess levels of neutralising antibodies across different severities of COVID-19 disease. By providing a method to calibrate most of the variants of concern so that the WHO International Standard for anti-SARS-CoV-2 Immunoglobulin reagent could be used to standardise our results, therefore making them comparable to other laboratories who also standardised their data in an identical manner. Implications of all the available evidence Continual use and accumulation of standardised data would eventually lead to defining the correlates of protection against SARS-CoV-2. This could help to inform medical staff to identify which individuals would be a greater risk of a potential reinfection to SARS-CoV-2.

ACS Style

Diego Cantoni; Martin Mayora-Neto; Angalee Nadesalingham; David A. Wells; George W. Carnell; Luis Ohlendorf; Matteo Ferarri; Phil Palmer; Andrew C.Y. Chan; Peter Smith; Emma M. Bentley; Sebastian Einhauser; Ralf Wagner; Mark Page; Gianmarco Raddi; Helen Baxendale; Javier Castillo-Olivares; Jonathan Heeney; Nigel Temperton. Neutralisation hierarchy of SARS-CoV-2 Variants of Concern using standardised, quantitative neutralisation assays reveals a correlation with disease severity; towards deciphering protective antibody thresholds. 2021, 1 .

AMA Style

Diego Cantoni, Martin Mayora-Neto, Angalee Nadesalingham, David A. Wells, George W. Carnell, Luis Ohlendorf, Matteo Ferarri, Phil Palmer, Andrew C.Y. Chan, Peter Smith, Emma M. Bentley, Sebastian Einhauser, Ralf Wagner, Mark Page, Gianmarco Raddi, Helen Baxendale, Javier Castillo-Olivares, Jonathan Heeney, Nigel Temperton. Neutralisation hierarchy of SARS-CoV-2 Variants of Concern using standardised, quantitative neutralisation assays reveals a correlation with disease severity; towards deciphering protective antibody thresholds. . 2021; ():1.

Chicago/Turabian Style

Diego Cantoni; Martin Mayora-Neto; Angalee Nadesalingham; David A. Wells; George W. Carnell; Luis Ohlendorf; Matteo Ferarri; Phil Palmer; Andrew C.Y. Chan; Peter Smith; Emma M. Bentley; Sebastian Einhauser; Ralf Wagner; Mark Page; Gianmarco Raddi; Helen Baxendale; Javier Castillo-Olivares; Jonathan Heeney; Nigel Temperton. 2021. "Neutralisation hierarchy of SARS-CoV-2 Variants of Concern using standardised, quantitative neutralisation assays reveals a correlation with disease severity; towards deciphering protective antibody thresholds." , no. : 1.

Review
Published: 31 January 2021 in Viruses
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Serological assays detecting neutralising antibodies are important for determining the immune responses following infection or vaccination and are also often considered a correlate of protection. The target of neutralising antibodies is usually located in the Envelope protein on the viral surface, which mediates cell entry. As such, presentation of the Envelope protein on a lentiviral particle represents a convenient alternative to handling of a potentially high containment virus or for those viruses with no established cell culture system. The flexibility, relative safety and, in most cases, ease of production of lentiviral pseudotypes, have led to their use in serological assays for many applications such as the evaluation of candidate vaccines, screening and characterization of anti-viral therapeutics, and sero-surveillance. Above all, the speed of production of the lentiviral pseudotypes, once the envelope sequence is published, makes them important tools in the response to viral outbreaks, as shown during the COVID-19 pandemic in 2020. In this review, we provide an overview of the landscape of the serological applications of pseudotyped lentiviral vectors, with a brief discussion on their production and batch quality analysis. Finally, we evaluate their role as surrogates for the real virus and possible alternatives.

ACS Style

Kamilla Toon; Emma Bentley; Giada Mattiuzzo. More Than Just Gene Therapy Vectors: Lentiviral Vector Pseudotypes for Serological Investigation. Viruses 2021, 13, 217 .

AMA Style

Kamilla Toon, Emma Bentley, Giada Mattiuzzo. More Than Just Gene Therapy Vectors: Lentiviral Vector Pseudotypes for Serological Investigation. Viruses. 2021; 13 (2):217.

Chicago/Turabian Style

Kamilla Toon; Emma Bentley; Giada Mattiuzzo. 2021. "More Than Just Gene Therapy Vectors: Lentiviral Vector Pseudotypes for Serological Investigation." Viruses 13, no. 2: 217.

Journal article
Published: 09 October 2020 in Viruses
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Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), responsible for the ongoing coronavirus disease (COVID-19) pandemic, is frequently shed in faeces during infection, and viral RNA has recently been detected in sewage in some countries. We have investigated the presence of SARS-CoV-2 RNA in wastewater samples from South-East England between 14th January and 12th May 2020. A novel nested RT-PCR approach targeting five different regions of the viral genome improved the sensitivity of RT-qPCR assays and generated nucleotide sequences at sites with known sequence polymorphisms among SARS-CoV-2 isolates. We were able to detect co-circulating virus variants, some specifically prevalent in England, and to identify changes in viral RNA sequences with time consistent with the recently reported increasing global dominance of Spike protein G614 pandemic variant. Low levels of viral RNA were detected in a sample from 11th February, 3 days before the first case was reported in the sewage plant catchment area. SARS-CoV-2 RNA concentration increased in March and April, and a sharp reduction was observed in May, showing the effects of lockdown measures. We conclude that viral RNA sequences found in sewage closely resemble those from clinical samples and that environmental surveillance can be used to monitor SARS-CoV-2 transmission, tracing virus variants and detecting virus importations.

ACS Style

Javier Martin; Dimitra Klapsa; Thomas Wilton; Maria Zambon; Emma Bentley; Erika Bujaki; Martin Fritzsche; Ryan Mate; Manasi Majumdar. Tracking SARS-CoV-2 in Sewage: Evidence of Changes in Virus Variant Predominance during COVID-19 Pandemic. Viruses 2020, 12, 1144 .

AMA Style

Javier Martin, Dimitra Klapsa, Thomas Wilton, Maria Zambon, Emma Bentley, Erika Bujaki, Martin Fritzsche, Ryan Mate, Manasi Majumdar. Tracking SARS-CoV-2 in Sewage: Evidence of Changes in Virus Variant Predominance during COVID-19 Pandemic. Viruses. 2020; 12 (10):1144.

Chicago/Turabian Style

Javier Martin; Dimitra Klapsa; Thomas Wilton; Maria Zambon; Emma Bentley; Erika Bujaki; Martin Fritzsche; Ryan Mate; Manasi Majumdar. 2020. "Tracking SARS-CoV-2 in Sewage: Evidence of Changes in Virus Variant Predominance during COVID-19 Pandemic." Viruses 12, no. 10: 1144.

Review
Published: 24 August 2019 in Viruses
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Following the Ebola outbreak in Western Africa in 2013-16, a global effort has taken place for preparedness for future outbreaks. As part of this response, the development of vaccines, treatments and diagnostic tools has been accelerated, especially towards pathogens listed as likely to cause an epidemic and for which there are no current treatments. Several of the priority pathogens identified by the World Health Organisation are haemorrhagic fever viruses. This review provides information on the role of reference materials as an enabling tool for the development and evaluation of assays, and ultimately vaccines and treatments. The types of standards available are described, along with how they can be applied for assay harmonisation through calibration as a relative potency to a common arbitrary unitage system (WHO International Unit). This assures that assay metrology is accurate and robust. We describe reference materials that have been or are being developed for haemorrhagic fever viruses and consider the issues surrounding their production, particularly that of biosafety where the viruses require specialised containment facilities. Finally, we advocate the use of reference materials at early stages, including research and development, as this helps produce reliable assays and can smooth the path to regulatory approval.

ACS Style

Giada Mattiuzzo; Emma M. Bentley; Mark Page. The Role of Reference Materials in the Research and Development of Diagnostic Tools and Treatments for Haemorrhagic Fever Viruses. Viruses 2019, 11, 781 .

AMA Style

Giada Mattiuzzo, Emma M. Bentley, Mark Page. The Role of Reference Materials in the Research and Development of Diagnostic Tools and Treatments for Haemorrhagic Fever Viruses. Viruses. 2019; 11 (9):781.

Chicago/Turabian Style

Giada Mattiuzzo; Emma M. Bentley; Mark Page. 2019. "The Role of Reference Materials in the Research and Development of Diagnostic Tools and Treatments for Haemorrhagic Fever Viruses." Viruses 11, no. 9: 781.

Journal article
Published: 28 April 2016 in New England Journal of Medicine
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The West African outbreak of Ebola virus disease that peaked in 2014 has caused more than 11,000 deaths. The development of an effective Ebola vaccine is a priority for control of a future outbreak. In this phase 1 study, we administered a single dose of the chimpanzee adenovirus 3 (ChAd3) vaccine encoding the surface glycoprotein of Zaire ebolavirus (ZEBOV) to 60 healthy adult volunteers in Oxford, United Kingdom. The vaccine was administered in three dose levels — 1×1010 viral particles, 2.5×1010 viral particles, and 5×1010 viral particles — with 20 participants in each group. We then assessed the effect of adding a booster dose of a modified vaccinia Ankara (MVA) strain, encoding the same Ebola virus glycoprotein, in 30 of the 60 participants and evaluated a reduced prime–boost interval in another 16 participants. We also compared antibody responses to inactivated whole Ebola virus virions and neutralizing antibody activity with those observed in phase 1 studies of a recombinant vesicular stomatitis virus–based vaccine expressing a ZEBOV glycoprotein (rVSV-ZEBOV) to determine relative potency and assess durability. No safety concerns were identified at any of the dose levels studied. Four weeks after immunization with the ChAd3 vaccine, ZEBOV-specific antibody responses were similar to those induced by rVSV-ZEBOV vaccination, with a geometric mean titer of 752 and 921, respectively. ZEBOV neutralization activity was also similar with the two vaccines (geometric mean titer, 14.9 and 22.2, respectively). Boosting with the MVA vector increased virus-specific antibodies by a factor of 12 (geometric mean titer, 9007) and increased glycoprotein-specific CD8+ T cells by a factor of 5. Significant increases in neutralizing antibodies were seen after boosting in all 30 participants (geometric mean titer, 139; P<0.001). Virus-specific antibody responses in participants primed with ChAd3 remained positive 6 months after vaccination (geometric mean titer, 758) but were significantly higher in those who had received the MVA booster (geometric mean titer, 1750; P<0.001). The ChAd3 vaccine boosted with MVA elicited B-cell and T-cell immune responses to ZEBOV that were superior to those induced by the ChAd3 vaccine alone. (Funded by the Wellcome Trust and others; ClinicalTrials.gov number, NCT02240875.)

ACS Style

Katie Ewer; Tommy Rampling; Navin Venkatraman; Georgina Bowyer; Danny Wright; Teresa Lambe; Egeruan B. Imoukhuede; Ruth Payne; Sarah Katharina Fehling; Thomas Strecker; Nadine Biedenkopf; Verena Krähling; Claire M. Tully; Nick Edwards; Emma M. Bentley; Dhanraj Samuel; Genevieve Labbe; Jing Jin; Malick Gibani; Alice Minhinnick; Morven Wilkie; Ian Poulton; Natalie Lella; Rachel Roberts; Felicity Hartnell; Carly Bliss; Kailan Sierra-Davidson; Jonathan Powlson; Eleanor Berrie; Richard Tedder; Francois Roman; Iris De Ryck; Alfredo Nicosia; Nancy J. Sullivan; Daphne A. Stanley; Olivier T. Mbaya; Julie E. Ledgerwood; Richard M. Schwartz; Loredana Siani; Stefano Colloca; Antonella Folgori; Stefania Di Marco; Riccardo Cortese; Edward Wright; Stephan Becker; Barney S. Graham; Richard A. Koup; Myron M. Levine; Ariane Volkmann; Paul Chaplin; Andrew J. Pollard; Simon Draper; W. Ripley Ballou; Alison Lawrie; Sarah C. Gilbert; Adrian V.S. Hill. A Monovalent Chimpanzee Adenovirus Ebola Vaccine Boosted with MVA. New England Journal of Medicine 2016, 374, 1635 -1646.

AMA Style

Katie Ewer, Tommy Rampling, Navin Venkatraman, Georgina Bowyer, Danny Wright, Teresa Lambe, Egeruan B. Imoukhuede, Ruth Payne, Sarah Katharina Fehling, Thomas Strecker, Nadine Biedenkopf, Verena Krähling, Claire M. Tully, Nick Edwards, Emma M. Bentley, Dhanraj Samuel, Genevieve Labbe, Jing Jin, Malick Gibani, Alice Minhinnick, Morven Wilkie, Ian Poulton, Natalie Lella, Rachel Roberts, Felicity Hartnell, Carly Bliss, Kailan Sierra-Davidson, Jonathan Powlson, Eleanor Berrie, Richard Tedder, Francois Roman, Iris De Ryck, Alfredo Nicosia, Nancy J. Sullivan, Daphne A. Stanley, Olivier T. Mbaya, Julie E. Ledgerwood, Richard M. Schwartz, Loredana Siani, Stefano Colloca, Antonella Folgori, Stefania Di Marco, Riccardo Cortese, Edward Wright, Stephan Becker, Barney S. Graham, Richard A. Koup, Myron M. Levine, Ariane Volkmann, Paul Chaplin, Andrew J. Pollard, Simon Draper, W. Ripley Ballou, Alison Lawrie, Sarah C. Gilbert, Adrian V.S. Hill. A Monovalent Chimpanzee Adenovirus Ebola Vaccine Boosted with MVA. New England Journal of Medicine. 2016; 374 (17):1635-1646.

Chicago/Turabian Style

Katie Ewer; Tommy Rampling; Navin Venkatraman; Georgina Bowyer; Danny Wright; Teresa Lambe; Egeruan B. Imoukhuede; Ruth Payne; Sarah Katharina Fehling; Thomas Strecker; Nadine Biedenkopf; Verena Krähling; Claire M. Tully; Nick Edwards; Emma M. Bentley; Dhanraj Samuel; Genevieve Labbe; Jing Jin; Malick Gibani; Alice Minhinnick; Morven Wilkie; Ian Poulton; Natalie Lella; Rachel Roberts; Felicity Hartnell; Carly Bliss; Kailan Sierra-Davidson; Jonathan Powlson; Eleanor Berrie; Richard Tedder; Francois Roman; Iris De Ryck; Alfredo Nicosia; Nancy J. Sullivan; Daphne A. Stanley; Olivier T. Mbaya; Julie E. Ledgerwood; Richard M. Schwartz; Loredana Siani; Stefano Colloca; Antonella Folgori; Stefania Di Marco; Riccardo Cortese; Edward Wright; Stephan Becker; Barney S. Graham; Richard A. Koup; Myron M. Levine; Ariane Volkmann; Paul Chaplin; Andrew J. Pollard; Simon Draper; W. Ripley Ballou; Alison Lawrie; Sarah C. Gilbert; Adrian V.S. Hill. 2016. "A Monovalent Chimpanzee Adenovirus Ebola Vaccine Boosted with MVA." New England Journal of Medicine 374, no. 17: 1635-1646.

Journal article
Published: 18 March 2016 in EMBO Molecular Medicine
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Currently available rabies post‐exposure prophylaxis (PEP) for use in humans includes equine or human rabies immunoglobulins (RIG). The replacement of RIG with an equally or more potent and safer product is strongly encouraged due to the high costs and limited availability of existing RIG. In this study, we identified two broadly neutralizing human monoclonal antibodies that represent a valid and affordable alternative to RIG in rabies PEP. Memory B cells from four selected vaccinated donors were immortalized and monoclonal antibodies were tested for neutralizing activity and epitope specificity. Two antibodies, identified as RVC20 and RVC58 (binding to antigenic site I and III, respectively), were selected for their potency and broad‐spectrum reactivity. In vitro, RVC20 and RVC58 were able to neutralize all 35 rabies virus (RABV) and 25 non‐RABV lyssaviruses. They showed higher potency and breath compared to antibodies under clinical development (namely CR57, CR4098, and RAB1) and commercially available human RIG. In vivo, the RVC20–RVC58 cocktail protected Syrian hamsters from a lethal RABV challenge and did not affect the endogenous hamster post‐vaccination antibody response.

ACS Style

Paola De Benedictis; Andrea Minola; Elena Rota Nodari; Roberta Aiello; Barbara Zecchin; Angela Salomoni; Mathilde Foglierini; Gloria Agatic; Fabrizia Vanzetta; Rachel Lavenir; Anthony Lepelletier; Emma M. Bentley; Robin Weiss; Giovanni Cattoli; Ilaria Capua; Federica Sallusto; Edward Wright; Antonio Lanzavecchia; Hervé Bourhy; Davide Corti. Development of broad‐spectrum human monoclonal antibodies for rabies post‐exposure prophylaxis. EMBO Molecular Medicine 2016, 8, 407 -421.

AMA Style

Paola De Benedictis, Andrea Minola, Elena Rota Nodari, Roberta Aiello, Barbara Zecchin, Angela Salomoni, Mathilde Foglierini, Gloria Agatic, Fabrizia Vanzetta, Rachel Lavenir, Anthony Lepelletier, Emma M. Bentley, Robin Weiss, Giovanni Cattoli, Ilaria Capua, Federica Sallusto, Edward Wright, Antonio Lanzavecchia, Hervé Bourhy, Davide Corti. Development of broad‐spectrum human monoclonal antibodies for rabies post‐exposure prophylaxis. EMBO Molecular Medicine. 2016; 8 (4):407-421.

Chicago/Turabian Style

Paola De Benedictis; Andrea Minola; Elena Rota Nodari; Roberta Aiello; Barbara Zecchin; Angela Salomoni; Mathilde Foglierini; Gloria Agatic; Fabrizia Vanzetta; Rachel Lavenir; Anthony Lepelletier; Emma M. Bentley; Robin Weiss; Giovanni Cattoli; Ilaria Capua; Federica Sallusto; Edward Wright; Antonio Lanzavecchia; Hervé Bourhy; Davide Corti. 2016. "Development of broad‐spectrum human monoclonal antibodies for rabies post‐exposure prophylaxis." EMBO Molecular Medicine 8, no. 4: 407-421.

Conference info
Published: 30 April 2015 in Vaccine
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The globalization of the world's economies, accompanied by increasing international travel, changing climates, altered human behaviour and demographics is leading to the emergence of different viral diseases, many of which are highly pathogenic and hence are considered of great public and animal health importance. To undertake basic research and therapeutic development, many of these viruses require handling by highly trained staff in BSL-3/4 facilities not readily available to the majority of the global R&D community. In order to circumvent the enhanced biosafety requirement, the development of non-pathogenic, replication-defective pseudotyped viruses is an effective and established solution to permit the study of many aspects of virus biology in a low containment biosafety level (BSL)-1/2 laboratory. Under the spectre of the unfolding Ebola crisis, this timely conference (the second to be organised by the Viral Pseudotype Unit, www.viralpseudotypeunit.info*) discusses the recent advances in pseudotype technology and how it is revolutionizing the study of important human and animal pathogens (human and avian influenza viruses, rabies/lyssaviruses, HIV, Marburg and Ebola viruses). Key topics addressed in this conference include the exploitation of pseudotypes for serology and serosurveillance, immunogenicity testing of current and next-generation vaccines and new pseudotype assay formats (multiplexing, kit development). *The first pseudotype-focused Euroscicon conference organised by the Viral Pseudotype Unit was recently reviewed [1].

ACS Style

Emma M. Bentley; Stuart T. Mather; Nigel J. Temperton. The use of pseudotypes to study viruses, virus sero-epidemiology and vaccination. Vaccine 2015, 33, 2955 -2962.

AMA Style

Emma M. Bentley, Stuart T. Mather, Nigel J. Temperton. The use of pseudotypes to study viruses, virus sero-epidemiology and vaccination. Vaccine. 2015; 33 (26):2955-2962.

Chicago/Turabian Style

Emma M. Bentley; Stuart T. Mather; Nigel J. Temperton. 2015. "The use of pseudotypes to study viruses, virus sero-epidemiology and vaccination." Vaccine 33, no. 26: 2955-2962.