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  1. Article ; Online: Measuring Hepatitis C Virus Envelopment by Using a Proteinase K Protection Assay.

    Roder, Allison E / Horner, Stacy M

    Methods in molecular biology (Clifton, N.J.)

    2018  Volume 1911, Page(s) 209–217

    Abstract: The infectious virion of hepatitis C virus (HCV) is made up of the viral nucleocapsid surrounded by an envelope that contains an ER-derived membrane bilayer, cellular lipids, and the viral E1 and E2 glycoproteins. Because the infectious HCV particle ... ...

    Abstract The infectious virion of hepatitis C virus (HCV) is made up of the viral nucleocapsid surrounded by an envelope that contains an ER-derived membrane bilayer, cellular lipids, and the viral E1 and E2 glycoproteins. Because the infectious HCV particle contains both protein and lipid layers, selective disruption of these layers and analysis for the presence or absence of resulting virion components can be used to study the virion assembly process. This chapter describes an experimental method to measure HCV virion envelopment, which can reveal the mechanisms of how specific viral protein-protein interactions and host factors contribute to the process of HCV envelopment.
    MeSH term(s) Cell Culture Techniques/methods ; Cell Line ; Electroporation/methods ; Endopeptidase K/metabolism ; Hepacivirus/genetics ; Hepacivirus/physiology ; Hepatitis C/virology ; Humans ; Immunoblotting/methods ; RNA, Viral/genetics ; Viral Core Proteins/genetics ; Viral Core Proteins/metabolism ; Virion/genetics ; Virion/physiology ; Virus Assembly
    Chemical Substances RNA, Viral ; Viral Core Proteins ; Endopeptidase K (EC 3.4.21.64)
    Language English
    Publishing date 2018-12-28
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-8976-8_14
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  2. Article ; Online: The acidic domain of the hepatitis C virus NS4A protein is required for viral assembly and envelopment through interactions with the viral E1 glycoprotein.

    Roder, Allison E / Vazquez, Christine / Horner, Stacy M

    PLoS pathogens

    2019  Volume 15, Issue 2, Page(s) e1007163

    Abstract: Hepatitis C virus (HCV) assembly and envelopment are coordinated by a complex protein interaction network that includes most of the viral structural and nonstructural proteins. While the nonstructural protein 4A (NS4A) is known to be important for viral ... ...

    Abstract Hepatitis C virus (HCV) assembly and envelopment are coordinated by a complex protein interaction network that includes most of the viral structural and nonstructural proteins. While the nonstructural protein 4A (NS4A) is known to be important for viral particle production, the specific function of NS4A in this process is not well understood. We performed mutagenesis of the C-terminal acidic domain of NS4A and found that mutation of several of these amino acids prevented the formation of the viral envelope, and therefore the production of infectious virions, without affecting viral RNA replication. In an overexpression system, we found that NS4A interacted with several viral proteins known to coordinate envelopment, including the viral E1 glycoprotein. One of the NS4A C-terminal mutations, Y45F, disrupted the interaction of NS4A with E1. Specifically, NS4A interacted with the first hydrophobic region of E1, a region previously described as regulating viral particle production. Indeed, we found that an E1 mutation in this region, D72A, also disrupted the interaction of NS4A with E1. Supernatants from HCV NS4A Y45F transfected cells had significantly reduced levels of HCV RNA, however they contained equivalent levels of Core protein. Interestingly, the Core protein secreted from these cells formed high order oligomers with a density matching the infectious virus secreted from wild-type cells. These results suggest that this Y45F mutation in NS4A causes secretion of low-density Core particles lacking genomic HCV RNA. These results corroborate previous findings showing that the E1 D72A mutation also causes secretion of Core complexes lacking genomic HCV RNA, and therefore suggest that the interaction between NS4A and E1 is involved in the incorporation of viral RNA into infectious HCV particles. Our findings define a new role for NS4A in the HCV lifecycle and help elucidate the protein interactions necessary for production of infectious virus.
    MeSH term(s) Amino Acid Sequence ; Base Sequence ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Cell Line ; Hepacivirus/genetics ; Hepacivirus/metabolism ; Hepacivirus/physiology ; Hepatitis C, Chronic/virology ; Humans ; Intracellular Signaling Peptides and Proteins ; Mutation ; Protein Domains ; RNA, Viral ; Viral Envelope Proteins/genetics ; Viral Envelope Proteins/metabolism ; Viral Nonstructural Proteins/genetics ; Viral Nonstructural Proteins/metabolism ; Virion/metabolism ; Virion/physiology ; Virus Assembly ; Virus Replication
    Chemical Substances Carrier Proteins ; E1 protein, Hepatitis C virus ; Intracellular Signaling Peptides and Proteins ; NS4A cofactor peptide, Hepatitis C virus ; RNA, Viral ; Viral Envelope Proteins ; Viral Nonstructural Proteins
    Language English
    Publishing date 2019-02-07
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2205412-1
    ISSN 1553-7374 ; 1553-7374
    ISSN (online) 1553-7374
    ISSN 1553-7374
    DOI 10.1371/journal.ppat.1007163
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  3. Article ; Online: Detection of SARS-CoV2 variants by Mesa Accula.

    Totten, Arthur H / Youn, Jung-Ho / Roder, Allison / Ghedin, Elodie / Palmore, Tara N / Frank, Karen M / Das, Sanchita / Zelazny, Adrian M

    Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology

    2021  Volume 141, Page(s) 104901

    MeSH term(s) COVID-19 ; Humans ; SARS-CoV-2
    Language English
    Publishing date 2021-06-19
    Publishing country Netherlands
    Document type Letter ; Research Support, N.I.H., Intramural
    ZDB-ID 1446080-4
    ISSN 1873-5967 ; 1386-6532
    ISSN (online) 1873-5967
    ISSN 1386-6532
    DOI 10.1016/j.jcv.2021.104901
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  4. Article ; Online: Cooperation between the Hepatitis C Virus p7 and NS5B Proteins Enhances Virion Infectivity.

    Aligeti, Mounavya / Roder, Allison / Horner, Stacy M

    Journal of virology

    2015  Volume 89, Issue 22, Page(s) 11523–11533

    Abstract: Unlabelled: The molecular mechanisms that govern hepatitis C virus (HCV) assembly, release, and infectivity are still not yet fully understood. In the present study, we sequenced a genotype 2A strain of HCV (JFH-1) that had been cell culture adapted in ... ...

    Abstract Unlabelled: The molecular mechanisms that govern hepatitis C virus (HCV) assembly, release, and infectivity are still not yet fully understood. In the present study, we sequenced a genotype 2A strain of HCV (JFH-1) that had been cell culture adapted in Huh-7.5 cells to produce nearly 100-fold-higher viral titers than the parental strain. Sequence analysis identified nine mutations in the genome, present within both the structural and nonstructural genes. The infectious clone of this virus containing all nine culture-adapted mutations had 10-fold-higher levels of RNA replication and RNA release into the supernatant but had nearly 1,000-fold-higher viral titers, resulting in an increased specific infectivity compared to wild-type JFH-1. Two mutations, identified in the p7 polypeptide and NS5B RNA-dependent RNA polymerase, were sufficient to increase the specific infectivity of JFH-1. We found that the culture-adapted mutation in p7 promoted an increase in the size of cellular lipid droplets following transfection of viral RNA. In addition, we found that the culture-adaptive mutations in p7 and NS5B acted synergistically to enhance the specific viral infectivity of JFH-1 by decreasing the level of sphingomyelin in the virion. Overall, these results reveal a genetic interaction between p7 and NS5B that contributes to virion specific infectivity. Furthermore, our results demonstrate a novel role for the RNA-dependent RNA polymerase NS5B in HCV assembly.
    Importance: Hepatitis C virus assembly and release depend on viral interactions with host lipid metabolic pathways. Here, we demonstrate that the viral p7 and NS5B proteins cooperate to promote virion infectivity by decreasing sphingomyelin content in the virion. Our data uncover a new role for the viral RNA-dependent RNA polymerase NS5B and p7 proteins in contributing to virion morphogenesis. Overall, these findings are significant because they reveal a genetic interaction between p7 and NS5B, as well as an interaction with sphingomyelin that regulates virion infectivity. Our data provide new strategies for targeting host lipid-virus interactions as potential targets for therapies against HCV infection.
    MeSH term(s) Base Sequence ; Cell Line, Tumor ; Hepacivirus/genetics ; Hepacivirus/metabolism ; Hepacivirus/pathogenicity ; Hepatitis C/virology ; Humans ; Lipid Droplets/physiology ; Lipoproteins, HDL/metabolism ; RNA, Viral/biosynthesis ; RNA, Viral/genetics ; RNA-Dependent RNA Polymerase/genetics ; RNA-Dependent RNA Polymerase/metabolism ; Sequence Analysis, RNA ; Sphingomyelins/metabolism ; Viral Load/genetics ; Viral Nonstructural Proteins/genetics ; Viral Nonstructural Proteins/metabolism ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Virion/genetics ; Virion/metabolism ; Virion/pathogenicity ; Virus Assembly/genetics ; Virus Replication/genetics
    Chemical Substances Lipoproteins, HDL ; RNA, Viral ; Sphingomyelins ; Viral Nonstructural Proteins ; Viral Proteins ; high density lipoprotein sphingomyelin ; p7 protein, Hepatitis C virus ; NS-5 protein, hepatitis C virus (EC 2.7.7.48) ; RNA-Dependent RNA Polymerase (EC 2.7.7.48)
    Language English
    Publishing date 2015-09-09
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.01185-15
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  5. Article ; Online: The impact of SARS-CoV-2 variants on the likelihood of children identified as sources of infection in the NIH workforce: a cohort study

    van Loben Sels, Jessica M. / Bailin, Heike B. / Bell, Michael R. / McCormick-Ell, Jessica / Das, Sanchita / Roder, Allison E. / Ghedin, Elodie / McGann, Michael / Castel, Amanda D. / Rebecca Prevots, D. / Kwan, Jennifer L.

    medRxiv

    Abstract: Abstract Background: Children (<18 years old) were not initially considered significant sources of infection (SOIs) for SARS-CoV-2. Risk mitigation strategies were thus prioritized for adults, and vaccination was inaccessible for children until mid-2021. ...

    Abstract Abstract Background: Children (<18 years old) were not initially considered significant sources of infection (SOIs) for SARS-CoV-2. Risk mitigation strategies were thus prioritized for adults, and vaccination was inaccessible for children until mid-2021. Emergence of novel variants led to significant increases in COVID-19 cases in both children and adults. Whether these emergence events and increased vulnerability of unvaccinated children had a synergistic effect resulting in increased caseloads in adults requires further exploration. Methods: A retrospective cohort study was conducted among 3,545 workers diagnosed with COVID-19. Case details were compiled during contact investigations. Variants of concern were identified following sequencing of biological samples collected through employer-based testing programs. Logistic regression was performed to compare the odds of having a child SOI based on the dominant variant in the workforce. Results: One-fourth (24.5%) of the cohort reported having a child in-residence; 11.2% identified a child as their SOI. In Alpha-dominant months, the odds of having a child SOI were 0.3, and the child SOI was likely older (5-17 years old). The odds of having a child SOI increased to 1.3 and 2.2 in Delta- and Omicron-dominant months, respectively. The odds of having younger child SOIs (<5 years old) were significantly higher in Omicron-dominant months. Conclusions: Children were highly likely to acquire the virus and posed a significant risk of transmission to their adult caretakers during Delta- and Omicron-dominant months. Without proper mitigation strategies in both the home and the workplace, child-associated transmission can threaten operations in the forms of staff shortages.
    Keywords covid19
    Language English
    Publishing date 2023-11-08
    Publisher Cold Spring Harbor Laboratory Press
    Document type Article ; Online
    DOI 10.1101/2023.11.07.23297422
    Database COVID19

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  6. Article ; Online: The impact of SARS-CoV-2 variants on the likelihood of children identified as sources of infection in the NIH workforce: a cohort study

    van Loben Sels, Jessica M / Bailin, Heike B / Bell, Michael R / McCormick-Ell, Jessica / McGann, Michael / Das, Sanchita / Roder, Allison E / Ghedin, Elodie / Castel, Amanda D / D. Rebecca, Prevots / Kwan, Jennifer L

    medRxiv

    Abstract: Abstract Background: Children (<18 years old) were not initially considered significant sources of infection (SOIs) for SARS-CoV-2. Risk mitigation strategies were thus prioritized for adults, and vaccination was inaccessible for children until mid-2021. ...

    Abstract Abstract Background: Children (<18 years old) were not initially considered significant sources of infection (SOIs) for SARS-CoV-2. Risk mitigation strategies were thus prioritized for adults, and vaccination was inaccessible for children until mid-2021. Emergence of novel variants led to significant increases in COVID-19 cases in both children and adults. Whether these emergence events and increased vulnerability of unvaccinated children had a synergistic effect resulting in increased caseloads in adults requires further exploration. Methods: A retrospective cohort study was conducted among 3,545 workers diagnosed with COVID-19. Case details were compiled during contact investigations. Variants of concern were identified following sequencing of biological samples collected through employer-based testing programs. Logistic regression was performed to compare the odds of having a child SOI based on the dominant variant in the workforce. Results: One-fourth (24.5%) of the cohort reported having a child in-residence; 11.2% identified a child as their SOI. In Alpha-dominant months, the odds of having a child SOI were 0.3, and the child SOI was likely older (5-17 years old). The odds of having a child SOI increased to 1.3 and 2.2 in Delta- and Omicron-dominant months, respectively. The odds of having younger child SOIs (<5 years old) were significantly higher in Omicron-dominant months. Conclusions: Children were highly likely to acquire the virus and posed a significant risk of transmission to their adult caretakers during Delta- and Omicron-dominant months. Without proper mitigation strategies in both the home and the workplace, child-associated transmission can threaten operations in the forms of staff shortages.
    Keywords covid19
    Language English
    Publishing date 2023-11-08
    Publisher Cold Spring Harbor Laboratory Press
    Document type Article ; Online
    DOI 10.1101/2023.11.07.23297422
    Database COVID19

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  7. Article ; Online: Accurate virus identification with interpretable Raman signatures by machine learning.

    Ye, Jiarong / Yeh, Yin-Ting / Xue, Yuan / Wang, Ziyang / Zhang, Na / Liu, He / Zhang, Kunyan / Ricker, RyeAnne / Yu, Zhuohang / Roder, Allison / Perea Lopez, Nestor / Organtini, Lindsey / Greene, Wallace / Hafenstein, Susan / Lu, Huaguang / Ghedin, Elodie / Terrones, Mauricio / Huang, Shengxi / Huang, Sharon Xiaolei

    Proceedings of the National Academy of Sciences of the United States of America

    2022  Volume 119, Issue 23, Page(s) e2118836119

    Abstract: Rapid identification of newly emerging or circulating viruses is an important first step toward managing the public health response to potential outbreaks. A portable virus capture device, coupled with label-free Raman spectroscopy, holds the promise of ... ...

    Abstract Rapid identification of newly emerging or circulating viruses is an important first step toward managing the public health response to potential outbreaks. A portable virus capture device, coupled with label-free Raman spectroscopy, holds the promise of fast detection by rapidly obtaining the Raman signature of a virus followed by a machine learning (ML) approach applied to recognize the virus based on its Raman spectrum, which is used as a fingerprint. We present such an ML approach for analyzing Raman spectra of human and avian viruses. A convolutional neural network (CNN) classifier specifically designed for spectral data achieves very high accuracy for a variety of virus type or subtype identification tasks. In particular, it achieves 99% accuracy for classifying influenza virus type A versus type B, 96% accuracy for classifying four subtypes of influenza A, 95% accuracy for differentiating enveloped and nonenveloped viruses, and 99% accuracy for differentiating avian coronavirus (infectious bronchitis virus [IBV]) from other avian viruses. Furthermore, interpretation of neural net responses in the trained CNN model using a full-gradient algorithm highlights Raman spectral ranges that are most important to virus identification. By correlating ML-selected salient Raman ranges with the signature ranges of known biomolecules and chemical functional groups—for example, amide, amino acid, and carboxylic acid—we verify that our ML model effectively recognizes the Raman signatures of proteins, lipids, and other vital functional groups present in different viruses and uses a weighted combination of these signatures to identify viruses.
    MeSH term(s) Disease Outbreaks ; Machine Learning ; Neural Networks, Computer ; Pandemics ; Serogroup ; Viruses/classification
    Language English
    Publishing date 2022-06-02
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Intramural
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.2118836119
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    Zs.A 1148: Show issues Location:
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  8. Article ; Online: Successful lung transplantation using an allograft from a COVID-19-recovered donor: a potential role for subgenomic RNA to guide organ utilization.

    Saharia, Kapil K / Ramelli, Sabrina C / Stein, Sydney R / Roder, Allison E / Kreitman, Allie / Banakis, Stephanie / Chung, Joon-Yong / Burbelo, Peter D / Singh, Manmeet / Reed, Robert M / Patel, Vipul / Rabin, Joseph / Krupnick, Alexander S / Cohen, Jeffrey I / de Wit, Emmie / Ghedin, Elodie / Hewitt, Stephen M / Vannella, Kevin M / Chertow, Daniel S /
    Grazioli, Alison

    American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

    2023  Volume 23, Issue 1, Page(s) 101–107

    Abstract: Although the risk of SARS-CoV-2 transmission through lung transplantation from acutely infected donors is high, the risks of virus transmission and long-term lung allograft outcomes are not as well described when using pulmonary organs from COVID-19- ... ...

    Abstract Although the risk of SARS-CoV-2 transmission through lung transplantation from acutely infected donors is high, the risks of virus transmission and long-term lung allograft outcomes are not as well described when using pulmonary organs from COVID-19-recovered donors. We describe successful lung transplantation for a COVID-19-related lung injury using lungs from a COVID-19-recovered donor who was retrospectively found to have detectable genomic SARS-CoV-2 RNA in the lung tissue by multiple highly sensitive assays. However, SARS-CoV-2 subgenomic RNA (sgRNA), a marker of viral replication, was not detectable in the donor respiratory tissues. One year after lung transplantation, the recipient has a good functional status, walking 1 mile several times per week without the need for supplemental oxygen and without any evidence of donor-derived SARS-CoV-2 transmission. Our findings highlight the limitations of current clinical laboratory diagnostic assays in detecting the persistence of SARS-CoV-2 RNA in the lung tissue. The persistence of SARS-CoV-2 RNA in the donor tissue did not appear to represent active viral replication via sgRNA testing and, most importantly, did not negatively impact the allograft outcome in the first year after lung transplantation. sgRNA is easily performed and may be a useful assay for assessing viral infectivity in organs from donors with a recent infection.
    MeSH term(s) Humans ; COVID-19 ; SARS-CoV-2/genetics ; Subgenomic RNA ; RNA, Viral/genetics ; Retrospective Studies ; Lung Transplantation ; Allografts
    Chemical Substances Subgenomic RNA ; RNA, Viral
    Language English
    Publishing date 2023-01-11
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ZDB-ID 2060594-8
    ISSN 1600-6143 ; 1600-6135
    ISSN (online) 1600-6143
    ISSN 1600-6135
    DOI 10.1016/j.ajt.2022.09.001
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  9. Article: Year-long COVID-19 infection reveals within-host evolution of SARS-CoV-2 in a patient with B cell depletion.

    Nussenblatt, Veronique / Roder, Allison E / Das, Sanchita / de Wit, Emmie / Youn, Jung-Ho / Banakis, Stephanie / Mushegian, Alexandra / Mederos, Christopher / Wang, Wei / Chung, Matthew / Pérez-Pérez, Lizzette / Palmore, Tara / Brudno, Jennifer N / Kochenderfer, James N / Ghedin, Elodie

    medRxiv : the preprint server for health sciences

    2021  

    Abstract: Background: B-cell depleting therapies may lead to protracted disease and prolonged viral shedding in individuals infected with SARS-CoV-2. Viral persistence in the setting of immunosuppression raises concern for viral evolution.: Methods: ... ...

    Abstract Background: B-cell depleting therapies may lead to protracted disease and prolonged viral shedding in individuals infected with SARS-CoV-2. Viral persistence in the setting of immunosuppression raises concern for viral evolution.
    Methods: Amplification of sub-genomic transcripts for the E gene (sgE) was done on nasopharyngeal samples over the course of 355 days in a patient infected with SARS-CoV-2 who had previously undergone CAR T cell therapy and had persistently positive SARS-CoV-2 nasopharyngeal swabs. Whole genome sequencing was performed on samples from the patient's original presentation and 10 months later.
    Results: Over the course of almost a year, the virus accumulated a unique in-frame deletion in the amino-terminal domain of the spike protein, and complete deletion of ORF7b and ORF8, the first report of its kind in an immunocompromised patient. Also, minority variants that were identified in the early samples-reflecting the heterogeneity of the initial infection-were found to be fixed late in the infection. Remdesivir and high-titer convalescent plasma treatment were given, and the infection was eventually cleared after 335 days of infection.
    Conclusions: The unique viral mutations found in this study highlight the importance of analyzing viral evolution in protracted SARS-CoV-2 infection, especially in immunosuppressed hosts, and the implication of these mutations in the emergence of viral variants.
    Summary: We report an immunocompromised patient with persistent symptomatic SARS-CoV-2 infection for 335 days. During this time, the virus accumulated a unique in-frame deletion in the spike, and a complete deletion of ORF7b and ORF8 which is the first report of its kind in an immunocompromised patient.
    Language English
    Publishing date 2021-10-05
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2021.10.02.21264267
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  10. Article ; Online: Yearlong COVID-19 Infection Reveals Within-Host Evolution of SARS-CoV-2 in a Patient With B-Cell Depletion.

    Nussenblatt, Veronique / Roder, Allison E / Das, Sanchita / de Wit, Emmie / Youn, Jung-Ho / Banakis, Stephanie / Mushegian, Alexandra / Mederos, Christopher / Wang, Wei / Chung, Matthew / Pérez-Pérez, Lizzette / Palmore, Tara / Brudno, Jennifer N / Kochenderfer, James N / Ghedin, Elodie

    The Journal of infectious diseases

    2021  Volume 225, Issue 7, Page(s) 1118–1123

    Abstract: B-cell-depleting therapies may lead to prolonged disease and viral shedding in individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and this viral persistence raises concern for viral evolution. We report sequencing of ... ...

    Abstract B-cell-depleting therapies may lead to prolonged disease and viral shedding in individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and this viral persistence raises concern for viral evolution. We report sequencing of early and late samples from a 335-day infection in an immunocompromised patient. The virus accumulated a unique deletion in the amino-terminal domain of the spike protein, and complete deletion of ORF7b and ORF8, the first report of its kind in an immunocompromised patient. Unique viral mutations found in this study highlight the importance of analyzing viral evolution in protracted SARS-CoV-2 infection, especially in immunosuppressed hosts.
    MeSH term(s) B-Lymphocytes ; COVID-19 ; Humans ; Immunocompromised Host ; SARS-CoV-2/genetics ; Spike Glycoprotein, Coronavirus/genetics ; Virus Shedding
    Chemical Substances Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
    Language English
    Publishing date 2021-12-23
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1093/infdis/jiab622
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