Publications by authors named "Gert-Jan Godeke"

18 Publications

  • Page 1 of 1

Validation and clinical evaluation of a SARS-CoV-2 surrogate virus neutralisation test (sVNT).

Emerg Microbes Infect 2020 Dec;9(1):2394-2403

Centre for Infectious Disease Control, WHO COVID-19 reference laboratory, RIVM, Bilthoven, Netherlands.

To understand SARS-CoV-2 immunity after natural infection or vaccination, functional assays such as virus neutralising assays are needed. So far, assays to detect SARS-CoV-2 neutralising antibodies rely on cell-culture based infection assays either using wild type SARS-CoV-2 or pseudotyped viruses. Such assays are labour-intensive, require appropriate biosafety facilities and are difficult to standardize. Recently, a new surrogate virus neutralisation test (sVNT) was described that uses the principle of an ELISA to measure the neutralisation capacity of anti-SARS-CoV-2 antibodies directed against the receptor binding domain. Here, we performed an independent evaluation of the robustness, specificity and sensitivity on an extensive panel of sera from 269 PCR-confirmed COVID-19 cases and 259 unmatched samples collected before 2020 and compared it to cell-based neutralisation assays. We found a high specificity of 99.2 (95%CI: 96.9-99.9) and overall sensitivity of 80.3 (95%CI: 74.9-84.8) for the sVNT. Clinical sensitivity increased between early (<14 days post symptom onset or post diagnosis, dpos/dpd) and late sera (>14 dpos/dpd) from 75.0 (64.7-83.2) to 83.1 (76.5-88.1). Also, higher severity was associated with an increase in clinical sensitivity. Upon comparison with cell-based neutralisation assays we determined an analytical sensitivity of 74.3 (56.4-86.9) and 98.2 (89.4-99.9) for titres ≥10 to <40 and ≥40 to <160, respectively. Only samples with a titre ≥160 were always positive in the sVNT. In conclusion, the sVNT can be used as an additional assay to determine the immune status of COVID-19 infected of vaccinated individuals but its value needs to be assessed for each specific context.
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http://dx.doi.org/10.1080/22221751.2020.1835448DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7605318PMC
December 2020

Accurate serology for SARS-CoV-2 and common human coronaviruses using a multiplex approach.

Emerg Microbes Infect 2020 Dec;9(1):1965-1973

Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands.

Serology is a crucial part of the public health response to the ongoing SARS-CoV-2 pandemic. Here, we describe the development, validation and clinical evaluation of a protein micro-array as a quantitative multiplex immunoassay that can identify S and N-directed SARS-CoV-2 IgG antibodies with high specificity and sensitivity and distinguish them from all currently circulating human coronaviruses. The method specificity was 100% for SARS-CoV-2 S1 and 96% for N antigen based on extensive syndromic (n=230 cases) and population panel (n=94) testing that also confirmed the high prevalence of seasonal human coronaviruses. To assess its potential role for both SARS-CoV-2 patient diagnostics and population studies, we evaluated a large heterogeneous COVID-19 cohort (n=330) and found an overall sensitivity of 89% (≥ 21 days post onset symptoms (dps)), ranging from 86% to 96% depending on severity of disease. For a subset of these patients longitudinal samples were provided up to 56 dps. Mild cases showed absent or delayed, and lower SARS-CoV-2 antibody responses. Overall, we present the development and extensive clinical validation of a multiplex coronavirus serological assay for syndromic testing, to answer research questions regarding to antibody responses, to support SARS-CoV-2 diagnostics and to evaluate epidemiological developments efficiently and with high-throughput.
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http://dx.doi.org/10.1080/22221751.2020.1813636DOI Listing
December 2020

Serological Evidence of MERS-CoV Antibodies in Dromedary Camels (Camelus dromedaries) in Laikipia County, Kenya.

PLoS One 2015 16;10(10):e0140125. Epub 2015 Oct 16.

Netherlands Centre for Infectious Disease Control, National Institute for Public Health and the Environment, Bilthoven, the Netherlands.

Middle East respiratory syndrome coronavirus (MERS-CoV) is a recently identified virus causing severe viral respiratory illness in people. Little is known about the reservoir in the Horn of Africa. In Kenya, where no human MERS cases have been reported, our survey of 335 dromedary camels, representing nine herds in Laikipia County, showed a high seroprevalence (46.9%) to MERS-CoV antibodies. Between herd differences were present (14.3%- 82.9%), but was not related to management type or herd isolation. Further research should focus on identifying similarity between MERS-CoV viral isolates in Kenya and clinical isolates from the Middle East and elsewhere.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0140125PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4608777PMC
June 2016

Occupational Exposure to Dromedaries and Risk for MERS-CoV Infection, Qatar, 2013-2014.

Emerg Infect Dis 2015 Aug;21(8):1422-5

We determined the presence of neutralizing antibodies to Middle East respiratory syndrome coronavirus in persons in Qatar with and without dromedary contact. Antibodies were only detected in those with contact, suggesting dromedary exposure as a risk factor for infection. Findings also showed evidence for substantial underestimation of the infection in populations at risk in Qatar.
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http://dx.doi.org/10.3201/eid2108.150481DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4517733PMC
August 2015

High proportion of MERS-CoV shedding dromedaries at slaughterhouse with a potential epidemiological link to human cases, Qatar 2014.

Infect Ecol Epidemiol 2015 15;5:28305. Epub 2015 Jul 15.

Department of Viroscience, Erasmus Medical Center, Rotterdam, The Netherlands.

Two of the earliest Middle East respiratory syndrome (MERS) cases were men who had visited the Doha central animal market and adjoining slaughterhouse in Qatar. We show that a high proportion of camels presenting for slaughter in Qatar show evidence for nasal MERS-CoV shedding (62/105). Sequence analysis showed the circulation of at least five different virus strains at these premises, suggesting that this location is a driver of MERS-CoV circulation and a high-risk area for human exposure. No correlation between RNA loads and levels of neutralizing antibodies was observed, suggesting limited immune protection and potential for reinfection despite previous exposure.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4505336PMC
http://dx.doi.org/10.3402/iee.v5.28305DOI Listing
July 2015

Spot the difference-development of a syndrome based protein microarray for specific serological detection of multiple flavivirus infections in travelers.

PLoS Negl Trop Dis 2015 Mar 13;9(3):e0003580. Epub 2015 Mar 13.

Erasmus Medical Centre, Viroscience Department, Rotterdam, The Netherlands.

Background: The family Flaviviridae, genus Flavivirus, holds many of the world's most prevalent arboviral diseases that are also considered the most important travel related arboviral infections. In most cases, flavivirus diagnosis in travelers is primarily based on serology as viremia is often low and typically has already been reduced to undetectable levels when symptoms set in and patients seek medical attention. Serological differentiation between flaviviruses and the false-positive results caused by vaccination and cross-reactivity among the different species, are problematic for surveillance and diagnostics of flaviviruses. Their partially overlapping geographic distribution and symptoms, combined with increase in travel, and preexisting antibodies due to flavivirus vaccinations, expand the need for rapid and reliable multiplex diagnostic tests to supplement currently used methods.

Goal: We describe the development of a multiplex serological protein microarray using recombinant NS1 proteins for detection of medically important viruses within the genus Flavivirus. Sera from clinical flavivirus patients were used for primary development of the protein microarray.

Results: Results show a high IgG and IgM sensitivity and specificity for individual NS1 antigens, and limited cross reactivity, even within serocomplexes. In addition, the serology based on this array allows for discrimination between infection and vaccination response for JEV vaccine, and no cross-reactivity with TBEV and YFV vaccine induced antibodies when testing for antibodies to other flaviviruses.

Conclusion: Based on these data, multiplex NS1-based protein microarray is a promising tool for surveillance and diagnosis of flaviviruses.
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http://dx.doi.org/10.1371/journal.pntd.0003580DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4359159PMC
March 2015

Geographic distribution of MERS coronavirus among dromedary camels, Africa.

Emerg Infect Dis 2014 Aug;20(8):1370-4

These authors contributed equally to this article.

We found serologic evidence for the circulation of Middle East respiratory syndrome coronavirus among dromedary camels in Nigeria, Tunisia, and Ethiopia. Circulation of the virus among dromedaries across broad areas of Africa may indicate that this disease is currently underdiagnosed in humans outside the Arabian Peninsula.
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http://dx.doi.org/10.3201/eid2008.140590DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4111168PMC
August 2014

Antibodies against MERS coronavirus in dromedary camels, United Arab Emirates, 2003 and 2013.

Emerg Infect Dis 2014 Apr;20(4):552-9

Middle East respiratory syndrome coronavirus (MERS-CoV) has caused an ongoing outbreak of severe acute respiratory tract infection in humans in the Arabian Peninsula since 2012. Dromedary camels have been implicated as possible viral reservoirs. We used serologic assays to analyze 651 dromedary camel serum samples from the United Arab Emirates; 151 of 651 samples were obtained in 2003, well before onset of the current epidemic, and 500 serum samples were obtained in 2013. Recombinant spike protein-specific immunofluorescence and virus neutralization tests enabled clear discrimination between MERS-CoV and bovine CoV infections. Most (632/651, 97.1%) camels had antibodies against MERS-CoV. This result included all 151 serum samples obtained in 2003. Most (389/651, 59.8%) serum samples had MERS-CoV-neutralizing antibody titers >1,280. Dromedary camels from the United Arab Emirates were infected at high rates with MERS-CoV or a closely related, probably conspecific, virus long before the first human MERS cases.
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http://dx.doi.org/10.3201/eid2004.131746DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3966379PMC
April 2014

Middle East respiratory syndrome coronavirus in dromedary camels: an outbreak investigation.

Lancet Infect Dis 2014 Feb 17;14(2):140-5. Epub 2013 Dec 17.

Department of Viroscience, Erasmus Medical Center, Rotterdam, Netherlands; Centre for Infectious Disease Research, Diagnostics and Screening, Division of Virology, National Institute for Public Health and the Environment, Bilthoven, Netherlands. Electronic address:

Background: Middle East respiratory syndrome coronavirus (MERS-CoV) causes severe lower respiratory tract infection in people. Previous studies suggested dromedary camels were a reservoir for this virus. We tested for the presence of MERS-CoV in dromedary camels from a farm in Qatar linked to two human cases of the infection in October, 2013.

Methods: We took nose swabs, rectal swabs, and blood samples from all camels on the Qatari farm. We tested swabs with RT-PCR, with amplification targeting the E gene (upE), nucleocapsid (N) gene, and open reading frame (ORF) 1a. PCR positive samples were tested by different MERS-CoV specific PCRs and obtained sequences were used for phylogentic analysis together with sequences from the linked human cases and other human cases. We tested serum samples from the camels for IgG immunofluorescence assay, protein microarray, and virus neutralisation assay.

Findings: We obtained samples from 14 camels on Oct 17, 2013. We detected MERS-CoV in nose swabs from three camels by three independent RT-PCRs and sequencing. The nucleotide sequence of an ORF1a fragment (940 nucleotides) and a 4·2 kb concatenated fragment were very similar to the MERS-CoV from two human cases on the same farm and a MERS-CoV isolate from Hafr-Al-Batin. Eight additional camel nose swabs were positive on one or more RT-PCRs, but could not be confirmed by sequencing. All camels had MERS-CoV spike-binding antibodies that correlated well with the presence of neutralising antibodies to MERS-CoV.

Interpretation: Our study provides virological confirmation of MERS-CoV in camels and suggests a recent outbreak affecting both human beings and camels. We cannot conclude whether the people on the farm were infected by the camels or vice versa, or if a third source was responsible.

Funding: European Union projects EMPERIE (contract number 223498), ANTIGONE (contract number 278976), and the VIRGO consortium.
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http://dx.doi.org/10.1016/S1473-3099(13)70690-XDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7106553PMC
February 2014

Middle East respiratory syndrome coronavirus neutralising serum antibodies in dromedary camels: a comparative serological study.

Lancet Infect Dis 2013 Oct 9;13(10):859-66. Epub 2013 Aug 9.

Centre for Infectious Disease Research, Diagnostics and Screening, Division Virology, National Institute for Public Health and the Environment, Bilthoven, Netherlands. Electronic address:

Background: A new betacoronavirus-Middle East respiratory syndrome coronavirus (MERS-CoV)-has been identified in patients with severe acute respiratory infection. Although related viruses infect bats, molecular clock analyses have been unable to identify direct ancestors of MERS-CoV. Anecdotal exposure histories suggest that patients had been in contact with dromedary camels or goats. We investigated possible animal reservoirs of MERS-CoV by assessing specific serum antibodies in livestock.

Methods: We took sera from animals in the Middle East (Oman) and from elsewhere (Spain, Netherlands, Chile). Cattle (n=80), sheep (n=40), goats (n=40), dromedary camels (n=155), and various other camelid species (n=34) were tested for specific serum IgG by protein microarray using the receptor-binding S1 subunits of spike proteins of MERS-CoV, severe acute respiratory syndrome coronavirus, and human coronavirus OC43. Results were confirmed by virus neutralisation tests for MERS-CoV and bovine coronavirus.

Findings: 50 of 50 (100%) sera from Omani camels and 15 of 105 (14%) from Spanish camels had protein-specific antibodies against MERS-CoV spike. Sera from European sheep, goats, cattle, and other camelids had no such antibodies. MERS-CoV neutralising antibody titres varied between 1/320 and 1/2560 for the Omani camel sera and between 1/20 and 1/320 for the Spanish camel sera. There was no evidence for cross-neutralisation by bovine coronavirus antibodies.

Interpretation: MERS-CoV or a related virus has infected camel populations. Both titres and seroprevalences in sera from different locations in Oman suggest widespread infection.

Funding: European Union, European Centre For Disease Prevention and Control, Deutsche Forschungsgemeinschaft.
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http://dx.doi.org/10.1016/S1473-3099(13)70164-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7106530PMC
October 2013

Detection of influenza A virus homo- and heterosubtype-specific memory B-cells using a novel protein microarray-based analysis tool.

J Med Virol 2013 May;85(5):899-909

Laboratory for Infectious Diseases and Screening, Center for Infectious Disease Control, Center for Infectious Disease Research, Diagnostics and Screening, National Institute for Public Health and the Environment, Bilthoven, the Netherlands.

The emergence of the A(H1N1) 2009 pandemic influenza virus was initially seen as a major world-wide health concern since a low degree of immunity to this virus strain was anticipated. However, age-specific infection attack rates and age-specific differences in seroresponse indicate that pre-existing immunity may have played a significant role in protection especially in older age groups. This study describes the use of a protein microarray as a multiplex analysis tool for detection of influenza virus H1 strain-specific memory B-cells before and after infection with A(H1N1)pdm09. The discrimination was based on detection of specific antibodies in culture supernatants from polyclonally stimulated B-cells against recombinant influenza virus HA1 proteins representing influenza virus subtypes H1 through H9. The protein microarray proved sensitive and specific for antibody detection in culture supernatants of B-cells, and with the potential to deduce a person's history of infection with particular influenza virus variants, including A(H1N1)pdm09. Blood samples obtained from different age groups prior to the pandemic in 2009 partly showed the presence of B-cells producing antibodies binding to the closely related A(H1N1) 1918 pandemic influenza virus, and of which the magnitude increased with age. These cross-reactive antibodies were produced by single memory B-cells present in these donors, and either bind to epitopes on HA1 which are shared within different H1 strains (homosubtypic response) or shared between different subtypes (heterosubtypic response).
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http://dx.doi.org/10.1002/jmv.23535DOI Listing
May 2013

Lack of evidence for zoonotic transmission of Schmallenberg virus.

Emerg Infect Dis 2012 Nov;18(11):1746-54

National Institute for Public Health and the Environment, Bilthoven, the Netherlands.

The emergence of Schmallenberg virus (SBV), a novel orthobunyavirus, in ruminants in Europe triggered a joint veterinary and public health response to address the possible consequences to human health. Use of a risk profiling algorithm enabled the conclusion that the risk for zoonotic transmission of SBV could not be excluded completely. Self-reported health problems were monitored, and a serologic study was initiated among persons living and/or working on SBV-affected farms. In the study set-up, we addressed the vector and direct transmission routes for putative zoonotic transfer. In total, 69 sheep farms, 4 goat farms, and 50 cattle farms were included. No evidence for SBV-neutralizing antibodies was found in serum of 301 participants. The lack of evidence for zoonotic transmission from either syndromic illness monitoring or serologic testing of presumably highly exposed persons suggests that the public health risk for SBV, given the current situation, is absent or extremely low.
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http://dx.doi.org/10.3201/eid1811.120650DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3559138PMC
November 2012

Come fly with me: review of clinically important arboviruses for global travelers.

J Clin Virol 2012 Nov 25;55(3):191-203. Epub 2012 Jul 25.

Centre for Infectious Disease Control, National Institute for Public Health and the Environment, P.O. Box 1, 3720 BA Bilthoven, The Netherlands.

Western tourists are increasingly traveling to exotic locations often located in tropical or subtropical regions of the world. The magnitude of international travel and the constantly changing dynamics of arbovirus diseases across the globe demand up-to-date information about arbovirus threats to travelers and the countries they visit. In this review, the current knowledge on arbovirus threats to global travelers is summarized and prioritized per region. Based on most common clinical syndromes, currently known arboviruses can be grouped to develop diagnostic algorithms to support decision-making in diagnostics. This review systematically combines and structures the current knowledge on medically important travel-related arboviruses and illustrates the necessity of a detailed patient history (travel history, symptoms experienced, vaccination history, engaged activities, tick or mosquito bite and use of repellent and onset of symptoms), to guide the diagnosis.
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http://dx.doi.org/10.1016/j.jcv.2012.07.004DOI Listing
November 2012

Quantitative performance of antibody array technology in a prenatal screening setting.

Clin Chem Lab Med 2011 Oct 28;50(2):325-32. Epub 2011 Oct 28.

Laboratory for Health Protection Research, National Institute of Public Health and the Environment, Bilthoven, The Netherlands.

Background: Antibody microarrays (Ab-array) represent a new, innovative proteomics platform for high-throughput protein expression profiling in body fluids. Because they allow for multiplexed measurements in small sample volumes, Ab-arrays are an interesting alternative to conventional indirect sandwich immunoassay (ELISA or DELFIA) tests in clinical or population screening if sets of markers are to be analyzed simultaneously. However, to allow implementation of Ab-arrays in clinical or population screening programs, it is of vital importance to establish that this method is both sensitive and quantitative.

Methods: This study developed and optimized a duplex Ab-array for pregnancy-associated plasma protein A (PAPP-A) and human chorion gonadotropin (fβ-hCG), two serum biomarkers currently analyzed by conventional biochemical techniques in prenatal screening. Serum samples from pregnant women, representing the dynamic range of both markers, were analyzed on Ab-arrays, and validated to the, in prenatal screening routinely applied, AutoDelfia system.

Results: Two different array hybridization conditions were tested, i.e., direct and indirect labeling, of which the indirect method displayed a sensitive and quantitative performance and a low intra- and inter-assay variation.

Conclusions: Taken together, these findings indicate that Ab-array technology is a promising alternative for ELISA or DELFIA in population screening programs, allowing future quantitative analysis of multiple biomarkers simultaneously in small volumes of serum.
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http://dx.doi.org/10.1515/CCLM.2011.767DOI Listing
October 2011

Syndromic surveillance in the Netherlands for the early detection of West Nile virus epidemics.

Vector Borne Zoonotic Dis 2006 ;6(2):161-9

Diagnostic Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, Bilthoven, The Netherlands.

West Nile virus (WNV) is an arthropod-borne flavivirus that is endemic in Africa, Europe, and Eastern Asia. The recent introduction and rapid dissemination of the virus in the United States as well as an increase in WNV outbreaks in Europe, has raised concerns for its spread in Europe. A surveillance system was developed to allow timely detection of an introduction of WNV infections in The Netherlands. This program focuses on cases presenting with neurological disease and includes the monitoring of hospital discharge diagnoses, trends in cerebrospinal fluid (CSF) diagnostic requests, laboratory testing of CSF, and monitoring of neurological disease in horses. Retrospective data from the hospital discharge records showed yearly peaks of unexplained meningitis and (meningo)encephalitis in the summer. A total of 781 CSF samples from humans and 71 serum and/or CSF samples from horses presenting with neurological disease of suspected viral etiology tested negative for the presence of specific antibodies to WNV. With a coverage rate of 59% in 2003, the probability that a cluster of five WNV cases presenting with neurological symptoms would have been detected was 99%. We conclude that, from 1999 to 2004, no evidence of WNV infection could be found in either humans or horses in The Netherlands.
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http://dx.doi.org/10.1089/vbz.2006.6.161DOI Listing
September 2006

Structural protein requirements in equine arteritis virus assembly.

J Virol 2004 Dec;78(23):13019-27

Virology Division, Department of Infectious Diseases and Immunology, Yalelaan 1, 3584 CL Utrecht, The Netherlands.

Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order Nidovirales. EAV particles contain seven structural proteins: the nucleocapsid protein N, the unglycosylated envelope proteins M and E, and the N-glycosylated membrane proteins GP(2b) (previously named G(S)), GP(3), GP(4), and GP(5) (previously named G(L)). Proteins N, M, and GP(5) are major virion components, E occurs in virus particles in intermediate amounts, and GP(4), GP(3), and GP(2b) are minor structural proteins. The M and GP(5) proteins occur in virus particles as disulfide-linked heterodimers while the GP(4), GP(3), and GP(2b) proteins are incorporated into virions as a heterotrimeric complex. Here, we studied the effect on virus assembly of inactivating the structural protein genes one by one in the context of a (full-length) EAV cDNA clone. It appeared that the three major structural proteins are essential for particle formation, while the other four virion proteins are dispensable. When one of the GP(2b), GP(3), or GP(4) proteins was missing, the incorporation of the remaining two minor envelope glycoproteins was completely blocked while that of the E protein was greatly reduced. The absence of E entirely prevented the incorporation of the GP(2b), GP(3), and GP(4) proteins into viral particles. EAV particles lacking GP(2b), GP(3), GP(4), and E did not markedly differ from wild-type virions in buoyant density, major structural protein composition, electron microscopic appearance, and genomic RNA content. On the basis of these results, we propose a model for the EAV particle in which the GP(2b)/GP(3)/GP(4) heterotrimers are positioned, in association with a defined number of E molecules, above the vertices of the putatively icosahedral nucleocapsid.
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http://dx.doi.org/10.1128/JVI.78.23.13019-13027.2004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC524988PMC
December 2004

[West Nile virus also in the Netherlands?].

Tijdschr Diergeneeskd 2004 Sep;129(17):561

Universiteit Utrecht.

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September 2004

Cleavage inhibition of the murine coronavirus spike protein by a furin-like enzyme affects cell-cell but not virus-cell fusion.

J Virol 2004 Jun;78(11):6048-54

Virology Division, Department of Infectious Diseases & Immunology, Yalelaan 1, 3584CL Utrecht, The Netherlands.

Cleavage of the mouse hepatitis coronavirus strain A59 spike protein was blocked in a concentration-dependent manner by a peptide furin inhibitor, indicating that furin or a furin-like enzyme is responsible for this process. While cell-cell fusion was clearly affected by preventing spike protein cleavage, virus-cell fusion was not, indicating that these events have different requirements.
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http://dx.doi.org/10.1128/JVI.78.11.6048-6054.2004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC415802PMC
June 2004