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  1. Article: Molecular Detection of SARS-CoV-2 in Formalin Fixed Paraffin Embedded Specimens.

    Liu, Jun / Babka, April M / Kearney, Brian J / Radoshitzky, Sheli R / Kuhn, Jens H / Zeng, Xiankun

    bioRxiv : the preprint server for biology

    2020  

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China in December 2019. The virus rapidly spread globally, resulting in a public-health crisis including more than one ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China in December 2019. The virus rapidly spread globally, resulting in a public-health crisis including more than one million cases and tens of thousands of deaths. Here, we describe the identification and evaluation of commercially available reagents and assays for the molecular detection of SARS-CoV-2 in infected formalin fixed paraffin embedded (FFPE) cell pellets. We identified a suitable rabbit polyclonal anti-SARS-CoV spike protein antibody and a mouse monoclonal anti-SARS-CoV nucleocapsid protein (NP) antibody for cross detection of the respective SARS-CoV-2 proteins by immunohistochemistry (IHC) and immunofluorescence assay (IFA). Next, we established RNAscope
    Keywords covid19
    Language English
    Publishing date 2020-04-21
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2020.04.21.042911
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Molecular detection of SARS-CoV-2 in formalin-fixed, paraffin-embedded specimens.

    Liu, Jun / Babka, April M / Kearney, Brian J / Radoshitzky, Sheli R / Kuhn, Jens H / Zeng, Xiankun

    JCI insight

    2020  Volume 5, Issue 12

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China, in December 2019. The virus rapidly spread globally, resulting in a public health crisis including almost 5 ... ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China, in December 2019. The virus rapidly spread globally, resulting in a public health crisis including almost 5 million cases and 323,256 deaths as of May 21, 2020. Here, we describe the identification and evaluation of commercially available reagents and assays for the molecular detection of SARS-CoV-2 in infected FFPE cell pellets. We identified a suitable rabbit polyclonal anti-SARS-CoV spike protein antibody and a mouse monoclonal anti-SARS-CoV nucleocapsid protein (NP) antibody for cross-detection of the respective SARS-CoV-2 proteins by IHC and immunofluorescence assay (IFA). Next, we established RNAscope in situ hybridization (ISH) to detect SARS-CoV-2 RNA. Furthermore, we established a multiplex FISH (mFISH) to detect positive-sense SARS-CoV-2 RNA and negative-sense SARS-CoV-2 RNA (a replicative intermediate indicating viral replication). Finally, we developed a dual staining assay using IHC and ISH to detect SARS-CoV-2 antigen and RNA in the same FFPE section. It is hoped that these reagents and assays will accelerate COVID-19 pathogenesis studies in humans and in COVID-19 animal models.
    MeSH term(s) Animals ; Antibodies, Viral/immunology ; Antigens, Viral/isolation & purification ; Betacoronavirus/genetics ; Betacoronavirus/immunology ; Betacoronavirus/isolation & purification ; COVID-19 ; COVID-19 Testing ; Clinical Laboratory Techniques/methods ; Coronavirus Infections/diagnosis ; Coronavirus Infections/pathology ; Coronavirus Infections/virology ; Formaldehyde ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Mice ; Nucleocapsid Proteins/immunology ; Pandemics ; Paraffin Embedding/methods ; Pathology, Molecular/methods ; Pneumonia, Viral/pathology ; Pneumonia, Viral/virology ; RNA, Viral/isolation & purification ; Rabbits ; SARS-CoV-2 ; Tissue Fixation/methods
    Chemical Substances Antibodies, Viral ; Antigens, Viral ; Nucleocapsid Proteins ; RNA, Viral ; Formaldehyde (1HG84L3525)
    Keywords covid19
    Language English
    Publishing date 2020-06-18
    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.
    ISSN 2379-3708
    ISSN (online) 2379-3708
    DOI 10.1172/jci.insight.139042
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Modeling the stability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on skin, currency, and clothing.

    Harbourt, David E / Haddow, Andrew D / Piper, Ashley E / Bloomfield, Holly / Kearney, Brian J / Fetterer, David / Gibson, Kathleen / Minogue, Timothy

    PLoS neglected tropical diseases

    2020  Volume 14, Issue 11, Page(s) e0008831

    Abstract: A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21st Century. Here, we ... ...

    Abstract A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21st Century. Here, we modeled stability of SARS-CoV-2 on skin, paper currency, and clothing to determine if these surfaces may factor in the fomite transmission dynamics of SARS-CoV-2. Skin, currency, and clothing samples were exposed to SARS-CoV-2 under laboratory conditions and incubated at three different temperatures (4°C± 2°C, 22°C± 2°C, and 37°C ± 2°C). We evaluated stability at 0 hours (h), 4 h, 8 h, 24 h, 72 h, 96 h, 7 days, and 14 days post-exposure. SARS-CoV-2 was stable on skin through the duration of the experiment at 4°C (14 days). Virus remained stable on skin for at least 96 h at 22°C and for at least 8h at 37°C. There were minimal differences between the tested currency samples. The virus remained stable on the $1 U.S.A. Bank Note for at least 96 h at 4°C while we did not detect viable virus on the $20 U.S.A. Bank Note samples beyond 72 h. The virus remained stable on both Bank Notes for at least 8 h at 22°C and 4 h at 37°C. Clothing samples were similar in stability to the currency. Viable virus remained for at least 96 h at 4°C and at least 4 h at 22°C. We did not detect viable virus on clothing samples at 37°C after initial exposure. This study confirms the inverse relationship between virus stability and temperature. Furthermore, virus stability on skin demonstrates the need for continued hand hygiene practices to minimize fomite transmission both in the general population as well as in workplaces where close contact is common.
    MeSH term(s) Betacoronavirus/physiology ; COVID-19 ; Clothing ; Coronavirus Infections/transmission ; Coronavirus Infections/virology ; Environmental Microbiology ; Humans ; Pandemics ; Pneumonia, Viral/transmission ; Pneumonia, Viral/virology ; SARS-CoV-2 ; Skin/virology ; Surface Properties ; Temperature
    Keywords covid19
    Language English
    Publishing date 2020-11-09
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2429704-5
    ISSN 1935-2735 ; 1935-2727
    ISSN (online) 1935-2735
    ISSN 1935-2727
    DOI 10.1371/journal.pntd.0008831
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Molecular detection of SARS-CoV-2 in formalin-fixed, paraffin-embedded specimens

    Liu, Jun / Babka, April M / Kearney, Brian J / Radoshitzky, Sheli R / Kuhn, Jens H / Zeng, Xiankun

    JCI insight

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China, in December 2019. The virus rapidly spread globally, resulting in a public health crisis including almost 5 ... ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of human coronavirus disease 2019 (COVID-19), emerged in Wuhan, China, in December 2019. The virus rapidly spread globally, resulting in a public health crisis including almost 5 million cases and 323,256 deaths as of May 21, 2020. Here, we describe the identification and evaluation of commercially available reagents and assays for the molecular detection of SARS-CoV-2 in infected FFPE cell pellets. We identified a suitable rabbit polyclonal anti-SARS-CoV spike protein antibody and a mouse monoclonal anti-SARS-CoV nucleocapsid protein (NP) antibody for cross-detection of the respective SARS-CoV-2 proteins by IHC and immunofluorescence assay (IFA). Next, we established RNAscope in situ hybridization (ISH) to detect SARS-CoV-2 RNA. Furthermore, we established a multiplex FISH (mFISH) to detect positive-sense SARS-CoV-2 RNA and negative-sense SARS-CoV-2 RNA (a replicative intermediate indicating viral replication). Finally, we developed a dual staining assay using IHC and ISH to detect SARS-CoV-2 antigen and RNA in the same FFPE section. It is hoped that these reagents and assays will accelerate COVID-19 pathogenesis studies in humans and in COVID-19 animal models.
    Keywords covid19
    Publisher WHO
    Document type Article
    Note WHO #Covidence: #215032
    Database COVID19

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  5. Article: Corning HYPERFlask® for viral amplification and production of diagnostic reagents

    Kearney, Brian J / Cynthia A. Rossi / Matthew A. Voorhees / Priscilla L. Williams / Randal J. Schoepp / Scott P. Olschner

    Journal of virological methods. 2017 Apr., v. 242

    2017  

    Abstract: Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or ... ...

    Abstract Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or roller bottle racks. The Corning HYPERFlask® is a multilayer flask that uses a gas permeable film to provide gas exchange between the cells and culture medium and the atmospheric environment. This study evaluated the suitability of the HYPERFlask for production of Lassa, Ebola, Bundibugyo, Reston, and Marburg viruses and compared it to more traditional methods using tissue culture flasks and roller bottles. The HYPERFlask produced cultures were equivalent in virus titer and indistinguishable in immunodiagnostic assays. The use of the Corning HYPERFlask for viral production is a viable alternative to traditional tissue culture flasks and roller bottles. HYPERFlasks allow for large volumes of virus to be produced in a small space without specialized equipment.
    Keywords bottles ; culture flasks ; culture media ; gas exchange ; research and development ; tissue culture ; viral load ; viruses
    Language English
    Dates of publication 2017-04
    Size p. 9-13.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 8013-5
    ISSN 1879-0984 ; 0166-0934
    ISSN (online) 1879-0984
    ISSN 0166-0934
    DOI 10.1016/j.jviromet.2016.12.011
    Database NAL-Catalogue (AGRICOLA)

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  6. Article ; Online: Modeling the stability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on skin, currency, and clothing

    Harbourt, David E. / Haddow, Andrew D. / Piper, Ashley E. / Bloomfield, Holly / Kearney, Brian J. / Fetterer, David / Gibson, Kathleen / Minogue, Timothy

    PLOS Neglected Tropical Diseases

    2020  Volume 14, Issue 11, Page(s) e0008831

    Abstract: A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21 st Century. Here, we ... ...

    Abstract A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21 st Century. Here, we modeled stability of SARS-CoV-2 on skin, paper currency, and clothing to determine if these surfaces may factor in the fomite transmission dynamics of SARS-CoV-2. Skin, currency, and clothing samples were exposed to SARS-CoV-2 under laboratory conditions and incubated at three different temperatures (4°C± 2°C, 22°C± 2°C, and 37°C ± 2°C). We evaluated stability at 0 hours (h), 4 h, 8 h, 24 h, 72 h, 96 h, 7 days, and 14 days post-exposure. SARS-CoV-2 was stable on skin through the duration of the experiment at 4°C (14 days). Virus remained stable on skin for at least 96 h at 22°C and for at least 8h at 37°C. There were minimal differences between the tested currency samples. The virus remained stable on the $1 U.S.A. Bank Note for at least 96 h at 4°C while we did not detect viable virus on the $20 U.S.A. Bank Note samples beyond 72 h. The virus remained stable on both Bank Notes for at least 8 h at 22°C and 4 h at 37°C. Clothing samples were similar in stability to the currency. Viable virus remained for at least 96 h at 4°C and at least 4 h at 22°C. We did not detect viable virus on clothing samples at 37°C after initial exposure. This study confirms the inverse relationship between virus stability and temperature. Furthermore, virus stability on skin demonstrates the need for continued hand hygiene practices to minimize fomite transmission both in the general population as well as in workplaces where close contact is common.
    Keywords Public Health, Environmental and Occupational Health ; Infectious Diseases ; covid19
    Language English
    Publisher Public Library of Science (PLoS)
    Publishing country us
    Document type Article ; Online
    ZDB-ID 2429704-5
    ISSN 1935-2735 ; 1935-2727
    ISSN (online) 1935-2735
    ISSN 1935-2727
    DOI 10.1371/journal.pntd.0008831
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

    Full text online

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  7. Article: Modeling the stability of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on skin, currency, and clothing

    Harbourt, David E / Haddow, Andrew D / Piper, Ashley E / Bloomfield, Holly / Kearney, Brian J / Fetterer, David / Gibson, Kathleen / Minogue, Timothy

    PLoS Negl Trop Dis

    Abstract: A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21st Century. Here, we ... ...

    Abstract A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21st Century. Here, we modeled stability of SARS-CoV-2 on skin, paper currency, and clothing to determine if these surfaces may factor in the fomite transmission dynamics of SARS-CoV-2. Skin, currency, and clothing samples were exposed to SARS-CoV-2 under laboratory conditions and incubated at three different temperatures (4°C± 2°C, 22°C± 2°C, and 37°C ± 2°C). We evaluated stability at 0 hours (h), 4 h, 8 h, 24 h, 72 h, 96 h, 7 days, and 14 days post-exposure. SARS-CoV-2 was stable on skin through the duration of the experiment at 4°C (14 days). Virus remained stable on skin for at least 96 h at 22°C and for at least 8h at 37°C. There were minimal differences between the tested currency samples. The virus remained stable on the $1 U.S.A. Bank Note for at least 96 h at 4°C while we did not detect viable virus on the $20 U.S.A. Bank Note samples beyond 72 h. The virus remained stable on both Bank Notes for at least 8 h at 22°C and 4 h at 37°C. Clothing samples were similar in stability to the currency. Viable virus remained for at least 96 h at 4°C and at least 4 h at 22°C. We did not detect viable virus on clothing samples at 37°C after initial exposure. This study confirms the inverse relationship between virus stability and temperature. Furthermore, virus stability on skin demonstrates the need for continued hand hygiene practices to minimize fomite transmission both in the general population as well as in workplaces where close contact is common.
    Keywords covid19
    Publisher WHO
    Document type Article
    Note WHO #Covidence: #917978
    Database COVID19

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  8. Article ; Online: Corning HYPERFlask

    Kearney, Brian J / Voorhees, Matthew A / Williams, Priscilla L / Olschner, Scott P / Rossi, Cynthia A / Schoepp, Randal J

    Journal of virological methods

    2017  Volume 242, Page(s) 9–13

    Abstract: Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or ... ...

    Abstract Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or roller bottle racks. The Corning HYPERFlask
    MeSH term(s) Animals ; Cercopithecus aethiops ; Culture Media ; Ebolavirus/growth & development ; Ebolavirus/isolation & purification ; Lassa virus/growth & development ; Lassa virus/isolation & purification ; Marburgvirus/growth & development ; Marburgvirus/isolation & purification ; Vero Cells ; Virus Cultivation/instrumentation ; Virus Cultivation/methods ; Virus Replication
    Chemical Substances Culture Media
    Language English
    Publishing date 2017-04
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 8013-5
    ISSN 1879-0984 ; 0166-0934
    ISSN (online) 1879-0984
    ISSN 0166-0934
    DOI 10.1016/j.jviromet.2016.12.011
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1565: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  9. Article ; Online: Sequence Optimized Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection of Crimean-Congo Hemorrhagic Fever Virus.

    Koehler, Jeffrey W / Delp, Korey L / Hall, Adrienne T / Olschner, Scott P / Kearney, Brian J / Garrison, Aura R / Altamura, Louis A / Rossi, Cynthia A / Minogue, Timothy D

    The American journal of tropical medicine and hygiene

    2018  Volume 98, Issue 1, Page(s) 211–215

    Abstract: Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus of the ... ...

    Abstract Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus of the genus
    MeSH term(s) DNA, Viral/genetics ; Hemorrhagic Fever Virus, Crimean-Congo/genetics ; Hemorrhagic Fever, Crimean/diagnosis ; Hemorrhagic Fever, Crimean/virology ; High-Throughput Nucleotide Sequencing/methods ; Humans ; Real-Time Polymerase Chain Reaction/methods ; Reverse Transcriptase Polymerase Chain Reaction/methods
    Chemical Substances DNA, Viral
    Language English
    Publishing date 2018-01-01
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2942-7
    ISSN 1476-1645 ; 0002-9637
    ISSN (online) 1476-1645
    ISSN 0002-9637
    DOI 10.4269/ajtmh.17-0165
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Ud II Zs.61: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

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  10. Article ; Online: Exposure Route Influences Disease Severity in the COVID-19 Cynomolgus Macaque Model.

    Bixler, Sandra L / Stefan, Christopher P / Jay, Alexandra N / Rossi, Franco D / Ricks, Keersten M / Shoemaker, Charles J / Moreau, Alicia M / Zeng, Xiankun / Hooper, Jay W / Dyer, David N / Frick, Ondraya M / Koehler, Jeffrey W / Kearney, Brian J / DiPinto, Nina / Liu, Jun / Tostenson, Samantha D / Clements, Tamara L / Smith, Jeffrey M / Johnson, Joshua A /
    Berrier, Kerry L / Esham, Heather L / Delp, Korey L / Coyne, Susan R / Bloomfield, Holly A / Kuehnert, Paul A / Akers, Kristen / Gibson, Kathleen M / Minogue, Timothy D / Nalca, Aysegul / Pitt, Margaret L M

    Viruses

    2022  Volume 14, Issue 5

    Abstract: The emergence of SARS-CoV-2 and the subsequent pandemic has highlighted the need for animal models that faithfully replicate the salient features of COVID-19 disease in humans. These models are necessary for the rapid selection, testing, and evaluation ... ...

    Abstract The emergence of SARS-CoV-2 and the subsequent pandemic has highlighted the need for animal models that faithfully replicate the salient features of COVID-19 disease in humans. These models are necessary for the rapid selection, testing, and evaluation of potential medical countermeasures. Here, we performed a direct comparison of two distinct routes of SARS-CoV-2 exposure-combined intratracheal/intranasal and small particle aerosol-in two nonhuman primate species, rhesus and cynomolgus macaques. While all four experimental groups displayed very few outward clinical signs, evidence of mild to moderate respiratory disease was present on radiographs and at necropsy. Cynomolgus macaques exposed via the aerosol route also developed the most consistent fever responses and had the most severe respiratory disease and pathology. This study demonstrates that while all four models produced suitable representations of mild COVID-like illness, aerosol exposure of cynomolgus macaques to SARS-CoV-2 produced the most severe disease, which may provide additional clinical endpoints for evaluating therapeutics and vaccines.
    MeSH term(s) Aerosols ; Animals ; COVID-19 ; Disease Models, Animal ; Macaca fascicularis ; SARS-CoV-2 ; Severity of Illness Index
    Chemical Substances Aerosols
    Language English
    Publishing date 2022-05-10
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2516098-9
    ISSN 1999-4915 ; 1999-4915
    ISSN (online) 1999-4915
    ISSN 1999-4915
    DOI 10.3390/v14051013
    Database MEDical Literature Analysis and Retrieval System OnLINE

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