Detection of #MERS-CoV #antigen on #formalin-fixed paraffin-embedded #nasal #tissue of #alpacas by immunohistochemistry using #human #mAbs directed against different epitopes of the spike protein (Vet Immunol Immunopathol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Vet Immunol Immunopathol. 2019 Sep 9;218:109939. doi: 10.1016/j.vetimm.2019.109939. [Epub ahead of print]

Detection of MERS-CoV antigen on formalin-fixed paraffin-embedded nasal tissue of alpacas by immunohistochemistry using human monoclonal antibodies directed against different epitopes of the spike protein.

Haverkamp AK1, Bosch BJ2, Spitzbarth I3, Lehmbecker A3, Te N4, Bensaid A4, Segalés J5, Baumgärtner W6.

Author information: 1 Department of Pathology, University of Veterinary Medicine Hannover Foundation, 30559 Hannover, Germany. 2 Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, 3584 CL Utrecht, the Netherlands. 3 Department of Pathology, University of Veterinary Medicine Hannover Foundation, 30559 Hannover, Germany; Center for Systems Neuroscience, 30559 Hannover, Germany. 4 IRTA, Centre de Recerca en Sanitat Animal (CReSA, IRTA-UAB), Campus de la Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain. 5 Departament de Sanitat i Anatomia Animals, Facultat de Veterinària, UAB, 08193 Bellaterra, Barcelona, Spain; UAB, Centre de Recerca en Sanitat Animal (CReSA, IRTA-UAB), Campus de la Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain. 6 Department of Pathology, University of Veterinary Medicine Hannover Foundation, 30559 Hannover, Germany; Center for Systems Neuroscience, 30559 Hannover, Germany. Electronic address: Wolfgang.Baumgaertner@tiho-hannover.de.

 

Abstract

Middle East respiratory syndrome (MERS) represents an important respiratory disease accompanied by lethal outcome in one third of human patients. In recent years, several investigators developed protective antibodies which could be used as prophylaxis in prospective human epidemics. In the current study, eight human monoclonal antibodies (mAbs) with neutralizing and non-neutralizing capabilities, directed against different epitopes of the MERS-coronavirus (MERS-CoV) spike (MERS-S) protein, were investigated with regard to their ability to immunohistochemically detect respective epitopes on formalin-fixed paraffin-embedded (FFPE) nasal tissue sections of MERS-CoV experimentally infected alpacas. The most intense immunoreaction was detected using a neutralizing antibody directed against the receptor binding domain S1B of the MERS-S protein, which produced an immunosignal in the cytoplasm of ciliated respiratory epithelium and along the apical membranous region. A similar staining was obtained by two other mAbs which recognize the sialic acid-binding domain and the ectodomain of the membrane fusion subunit S2, respectively. Five mAbs lacked immunoreactivity for MERS-CoV antigen on FFPE tissue, even though they belong, at least in part, to the same epitope group. In summary, three tested human mAbs demonstrated capacity for detection of MERS-CoV antigen on FFPE samples and may be implemented in double or triple immunohistochemical methods.

Copyright © 2019 Elsevier B.V. All rights reserved.

KEYWORDS: Immunohistochemistry; Middle East respiratory syndrome coronavirus; Monoclonal human antibodies; Spike protein

PMID: 31526954 DOI: 10.1016/j.vetimm.2019.109939

Keywords: MERS-CoV; Monoclonal antibodies; Diagnostic tests.

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Comparison of phenotypic and genotypic #diagnosis of acute #human #bocavirus 1 #infection in #children (J Clin Virol., abstract)

[Source: Journal of Clinical Virology, full page: (LINK). Abstract, edited.]

Journal of Clinical Virology / Available online 4 September 2019 / In Press, Journal Pre-proof / Short communication

Comparison of phenotypic and genotypic diagnosis of acute human bocavirus 1 infection in children

Nicola Isabelle Kols a,b, Heli Aatol a,c, Ville Peltol a,d, Man Xue Zaig a, Nora-Krukle e,f, Klaus Hedman e,g, Aurelija Zvirbliene h, Hanna Toivol a, c, Tytti Vuorinen i, Juha  M. Koskinen c, Andrea H. L. Bruning j, Andreas Christensen a,b, Maria Söderlund-Venermo e, Janne O. Koskinen c

{a} Department of Medical Microbiology, Clinic of Laboratory Medicine, St. Olavs Hospital HF, Trondheim University Hospital, 7006, Trondheim, Norway; {b} Department of Clinical and Molecular Medicine, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, 7491, Trondheim, Norway; {c} ArcDia International Oy Ltd, 20521, Turku, Finland; {d} Department of Paediatrics and Adolescent Medicine, Turku University Hospital and University of Turku, 20521, Turku, Finland; {e} Department of Virology, University of Helsinki, and Helsinki University Hospital, 00290, Helsinki, Finland; {f} Institute of Microbiology and Virology, Riga Stradins University, LV-1067, Riga, Latvia; {g} Helsinki University Hospital, 00029, Helsinki, Finland; {h} Department of Immunology and Cell Biology, Institute of Biotechnology, Life Sciences Center, Vilnius University, LT-10223, Vilnius, Lithuania; {i}
Department of Clinical Microbiology, Turku University Hospital and Department of Virology, University of Turku, 20521, Turku, Finland; {j} Department of Pediatric Infectious Diseases, Emma Children’s Hospital, Academic Medical Center, 1105, Amsterdam, the Netherlands

Received 26 February 2019, Revised 5 August 2019, Accepted 3 September 2019, Available online 4 September 2019. DOI: https://doi.org/10.1016/j.jcv.2019.09.003

 

Highlights

  • Diagnosis of HBoV1 has been based on detection of DNA or mRNA.
  • Rapid HBoV1 antigen detection is beneficial for diagnosing acute HBoV1 infections.
  • HBoV1 antigen detection is attractive for point-of-care use.

 

Keywords: Bocavirus; Diagnostic tests.

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Comparison of #Serologic #Assays for #MERS #Coronavirus (Emerg Infect Dis., abstract)

[Source: US Centers for Disease Control and Prevention (CDC), Emerging Infectious Diseases Journal, full page: (LINK). Abstract, edited.]

Volume 25, Number 10—October 2019 / Research

Comparison of Serologic Assays for Middle East Respiratory Syndrome Coronavirus

Ruth Harvey, Giada Mattiuzzo  , Mark Hassall, Andrea Sieberg, Marcel A. Müller, Christian Drosten, Peter Rigsby, Christopher J. Oxenford, and study participants

Author affiliations: National Institute for Biological Standards and Control—MHRA, Potters Bar, UK (R. Harvey, G. Mattiuzzo, M. Hassall, P. Rigsby); Charité-Universitätsmedizin Berlin, Berlin, Germany (A. Sieberg, M.A. Müller, C. Drosten); Humboldt-Universität zu Berlin, Berlin (A. Sieberg, M.A. Müller, C. Drosten); Berlin Institute of Health, Berlin (A. Sieberg, M.A. Müller, C. Drosten); German Centre for Infection Research, Berlin (M.A. Müller, C. Drosten); World Health Organization, Lyon, France (C.J. Oxenford).

 

Abstract

Middle East respiratory syndrome coronavirus (MERS-CoV) was detected in humans in 2012. Since then, sporadic outbreaks with primary transmission through dromedary camels to humans and outbreaks in healthcare settings have shown that MERS-CoV continues to pose a threat to human health. Several serologic assays for MERS-CoV have been developed globally. We describe a collaborative study to investigate the comparability of serologic assays for MERS-CoV and assess any benefit associated with the introduction of a standard reference reagent for MERS-CoV serology. Our study findings indicate that, when possible, laboratories should use a testing algorithm including >2 tests to ensure correct diagnosis of MERS-CoV. We also demonstrate that the use of a reference reagent greatly improves the agreement between assays, enabling more consistent and therefore more meaningful comparisons between results.

Keywords: MERS-CoV; Serology; Diagnostic tests.

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#Metagenomic next-generation #sequencing aids the #diagnosis of #viral #infections in febrile returning #travellers (J Infect., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

J Infect. 2019 Aug 6. pii: S0163-4453(19)30242-7. doi: 10.1016/j.jinf.2019.08.003. [Epub ahead of print]

Metagenomic next-generation sequencing aids the diagnosis of viral infections in febrile returning travellers.

Jerome H1, Taylor C2, Sreenu VB1, Klymenko T1, Jackson C3, Davis C1, Ashraf S1, Wilson-Davies E3, Jesudason N4, Devine K2, Harder L1, Aitken C3, Gunson R3, Thomson EC5.

Author information: 1 MRC-University of Glasgow Centre for Virus Research, 464 Bearsden Road, Glasgow, UK G61 1QH. 2 Department of Infectious Diseases, Queen Elizabeth University Hospital, 1345 Govan Rd, Govan, Glasgow, UK G51 4TF. 3 West of Scotland Specialist Virology Centre, Level 5, New Lister Building, Glasgow Royal Infirmary, 10-16 Alexandra Parade, Glasgow, UK G31 2ER. 4 Queen Elizabeth University Hospital, 1345 Govan Rd, Govan, Glasgow, UK G51 4TF. 5 MRC-University of Glasgow Centre for Virus Research, 464 Bearsden Road, Glasgow, UK G61 1QH; Department of Infectious Diseases, Queen Elizabeth University Hospital, 1345 Govan Rd, Govan, Glasgow, UK G51 4TF. Electronic address: emma.thomson@glasgow.ac.uk.

 

Abstract

OBJECTIVES:

Travel-associated infections are challenging to diagnose because of the broad spectrum of potential aetiologies. As a proof-of-principle study, we used MNGS to identify viral pathogens in clinical samples from returning travellers in a single centre to explore its suitability as a diagnostic tool.

METHODS:

Plasma samples from 40 returning travellers presenting with a fever of ≥38°C were retrospectively sequenced using MNGS on the Illumina MiSeq platform and compared with standard-of-care diagnostic assays.

RESULTS:

In total, 11/40 patients were diagnosed with a viral infection. Standard of care diagnostics revealed 5 viral infections using plasma samples; dengue virus 1 (n=2), hepatitis E (n=1), Ebola virus (n=1) and hepatitis A (n=1), all of which were detected by MNGS. Three additional patients with Chikungunya virus (n=2) and mumps virus were diagnosed by MNGS only. Respiratory infections detected by nasal/throat swabs only were not detected by MNGS of plasma. One patient had infection with malaria and mumps virus during the same admission.

CONCLUSIONS:

MNGS analysis of plasma samples improves the sensitivity of diagnosis of viral infections and has potential as an all-in-one diagnostic test. It can be used to identify infections that have not been considered by the treating physician, co-infections and new or emerging pathogens.

SUMMARY:

Next generation sequencing (NGS) has potential as an all-in-one diagnostic test. In this study we used NGS to diagnose returning travellers with acute febrile illness in the UK, highlighting cases where the diagnosis was missed using standard methods.

Copyright © 2019. Published by Elsevier Ltd.

KEYWORDS: Diagnosis; NGS; Returning travellers

PMID: 31398374 DOI: 10.1016/j.jinf.2019.08.003

Keywords: Infectious Diseases; Diagnostic tests.

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Optical #DNA #Mapping Combined with #Cas9-Targeted #Resistance #Gene #Identification for Rapid #Tracking of Resistance #Plasmids in a #NICU #Outbreak (mBio, abstract)

[Source: mBio, full page: (LINK). Abstract, edited.]

Optical DNA Mapping Combined with Cas9-Targeted Resistance Gene Identification for Rapid Tracking of Resistance Plasmids in a Neonatal Intensive Care Unit Outbreak

Santosh K. Bikkarolla, Viveka Nordberg, Fredrika Rajer, Vilhelm Müller, Muhammad Humaun Kabir, Sriram KK, Albertas Dvirnas, Tobias Ambjörnsson, Christian G. Giske, Lars Navér,Linus Sandegren, Fredrik Westerlund

Spyros Pournaras, Invited Editor, Karen Bush, Editor

DOI: 10.1128/mBio.00347-19

 

ABSTRACT

The global spread of antibiotic resistance among Enterobacteriaceae is largely due to multidrug resistance plasmids that can transfer between different bacterial strains and species. Horizontal gene transfer of resistance plasmids can complicate hospital outbreaks and cause problems in epidemiological tracing, since tracing is usually based on bacterial clonality. We have developed a method, based on optical DNA mapping combined with Cas9-assisted identification of resistance genes, which is used here to characterize plasmids during an extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae outbreak at a Swedish neonatal intensive care unit. The outbreak included 17 neonates initially colonized with ESBL-producing Klebsiella pneumoniae (ESBL-KP), some of which were found to carry additional ESBL-producing Escherichia coli (ESBL-EC) in follow-up samples. We demonstrate that all ESBL-KP isolates contained two plasmids with the blaCTX-M-15 gene located on the smaller one (~80 kbp). The same ESBL-KP clone was present in follow-up samples for up to 2 years in some patients, and the plasmid carrying the blaCTX-M-15 gene was stable throughout this time period. However, extensive genetic rearrangements within the second plasmid were observed in the optical DNA maps for several of the ESBL-KP isolates. Optical mapping also demonstrated that even though other bacterial clones and species carrying blaCTX-M group 1 genes were found in some neonates, no transfer of resistance plasmids had occurred. The data instead pointed toward unrelated acquisition of ESBL-producing Enterobacteriaceae (EPE). In addition to revealing important information about the specific outbreak, the method presented is a promising tool for surveillance and infection control in clinical settings.

IMPORTANCE

This study presents how a novel method, based on visualizing single plasmids using sequence-specific fluorescent labeling, could be used to analyze the genetic dynamics of an outbreak of resistant bacteria in a neonatal intensive care unit at a Swedish hospital. Plasmids are a central reason for the rapid global spread of bacterial resistance to antibiotics. In a single experimental procedure, this method replaces many traditional plasmid analysis techniques that together provide limited details and are slow to perform. The method is much faster than long-read whole-genome sequencing and offers direct genetic comparison of patient samples. We could conclude that no transfer of resistance plasmids had occurred between different bacteria during the outbreak and that secondary cases of ESBL-producing Enterobacteriaceae carriage were instead likely due to influx of new strains. We believe that the method offers potential in improving surveillance and infection control of resistant bacteria in hospitals.

Keywords: Antibiotics; Drugs Resistance; Enterobacteriaceae; Beta-lactams; Nosocomial Outbreaks; Diagnostic tests.

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Ensuring On-site #Ebola #Patient #Monitoring and Follow-up: #Development of a #Laboratory Structure Embedded in an #ETC (Disaster Med Public Health Prep., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Disaster Med Public Health Prep. 2019 Jun 24:1-7. doi: 10.1017/dmp.2019.39. [Epub ahead of print]

Ensuring On-site Ebola Patient Monitoring and Follow-up: Development of a Laboratory Structure Embedded in an Ebola Treatment Center.

Williams A1, Amand M2, Van den Bergh R1, De Clerck H3, Antierens A3, Chaillet P3.

Author information: 1 Médecins Sans Frontières,Operational Centre Brussels, Operational Research Unit LuxOR,Luxembourg, Luxembourg. 2 Médecins Sans Frontières,Operational Centre Brussels,Guinea Mission, Conakry,Guinea. 3 Médecins Sans Frontières,Operational Centre Brussels,Medical Department, Brussels,Belgium.

 

Abstract

The capacity to rapidly distinguish Ebola virus disease from other infectious diseases and to monitor biochemistry and viremia levels is crucial to the clinical management of suspected Ebola virus disease cases. This article describes the design and practical considerations of a laboratory straddling the high- and low-risk zones of an Ebola treatment center to produce timely diagnostic and clinical results for informed case management of Ebola virus disease in real-life conditions. This innovation may be of relevance for actors requiring flexible laboratory implementation in contexts of high-communicability, high-lethality disease outbreaks.

KEYWORDS: Ebola virus; disease outbreaks; laboratories; outbreak response; patient monitoring

PMID: 31232266 DOI: 10.1017/dmp.2019.39

Keywords: Ebola; Diagnostic tests.

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Development of a Specific #CHIKV-E2 #Monoclonal #Antibody for #Chikungunya #Diagnosis (Virol Sin., abstract)

[Source: Virologica Sinica, full page: (LINK). Abstract, edited.]

Development of a Specific CHIKV-E2 Monoclonal Antibody for Chikungunya Diagnosis

Authors: Jaemoo Kim, Jihyun Yang, Young Bong Kim, Hee-Jung Lee, Sehyun Kim, Haryoung Poo

Research Article / First Online: 18 June 2019

 

Abstract

Chikungunya fever is a vector-borne viral disease transmitted to humans by chikungunya virus (CHIKV)-infected mosquitoes. There have been many outbreaks of CHIKV infection worldwide, and the virus poses ongoing risks to global health. To prevent and control CHIKV infection, it is important to improve the current CHIKV diagnostic approaches to allow for the detection of low CHIKV concentrations and to correctly distinguish CHIKV infections from those due to other mosquito-transmitted viruses, including dengue virus (DENV), Japanese encephalitis virus (JEV), and Zika virus (ZIKV). Here, we produced monoclonal antibodies (mAbs) against the CHIKV envelope 2 protein (CHIKV-E2) and compared their sensitivity and specificity with commercially available mAbs using enzyme-linked immunosorbent assays (ELISA). Two anti-CHIKV-E2 mAbs, 19-1 and 21-1, showed higher binding affinities to CHIKV-E2 protein than the commercial mAbs did. In particular, the 19-1 mAb had the strongest binding affinity to inactivated CHIKV. Moreover, the 19-1 mAb had very little cross-reactivity with other mosquito-borne viruses, such as ZIKV, JEV, and DENV. These results suggest that the newly produced anti-CHIKV-E2 mAb, 19-1, could be used for CHIKV diagnostic approaches.

Keywords: Chikungunya virus – (CHIKV) Envelope 2 – Monoclonal antibody – Diagnosis – Sensitivity – Specificity

 

Notes

Acknowledgements

This work was supported by Grants from the R&D Convergence Program of National Research Council of Science & Technology (No. CAP-16-02-KIST) and the National Research Foundation of Korea (No. NRF-2016M3A9B6918584).

Author Contributions

HP designed the experiments; JK performed the experiments; SK, HL, and YK contributed to analyze cross-reactivities of anti-CHIKV-E2 antibodies to arboviruses; JK and JY analyzed the experiments and drafted the manuscript; HP supervised the experiments, analyzed results, and wrote the manuscript. All authors approved the final manuscript.

 

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no competing interests.

Animal and Human Rights Statement

Animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Korea Research Institute of Bioscience and Biotechnology (KRIBB) and performed according to the Guidelines for Animal Experiments of the KRIBB.

Keywords: Chikungunya fever; Monoclonal antibodies; Diagnostic tests.

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