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  1. Article: Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.

    Banik, Sukalyani / Saibire, Kaheerman / Suryavanshi, Shraddha / Johns, Glenn / Chakravorty, Soumitesh / Kwiatkowski, Robert / Alland, David / Banada, Padmapriya

    medRxiv : the preprint server for health sciences

    2021  

    Abstract: ... inactivation medium for downstream RT-PCR testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 ... and processing of clinical samples for RT-PCR based SARS-CoV-2 detection. ... Background: Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and ...

    Abstract Background: Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings.
    Methods: We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream RT-PCR testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C.
    Results: SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log
    Conclusion: eNAT and similar guanidinium thiocyanate-based media may be of value for transport, preservation, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection.
    Language English
    Publishing date 2021-01-20
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2021.01.15.21249891
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.

    Banik, Sukalyani / Saibire, Kaheerman / Suryavanshi, Shraddha / Johns, Glenn / Chakravorty, Soumitesh / Kwiatkowski, Robert / Alland, David / Banada, Padmapriya P

    PloS one

    2021  Volume 16, Issue 6, Page(s) e0252687

    Abstract: ... for transport, stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection. ... Background: Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and ... present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 ...

    Abstract Background: Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings.
    Methods: We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C.
    Results: SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log10 PFU/ml within a minute of incubation. When saliva was diluted 1:1 in eNAT, no cytopathic effect (CPE) on VeroE6 cells was observed, although SARS-CoV-2 RNA could be detected even after 30 min incubation and after two cell culture passages. A 1:2 (saliva:eNAT) dilution abrogated both CPE and detectable viral RNA after as little as 5 min incubation in eNAT. SARS-CoV-2 RNA from virus spiked at 5X the limit of detection remained positive up to 7 days of incubation in all tested conditions.
    Conclusion: eNAT and similar guanidinium thiocyanate-based media may be of value for transport, stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection.
    MeSH term(s) Animals ; COVID-19/diagnosis ; COVID-19/virology ; COVID-19 Nucleic Acid Testing/methods ; Chlorocebus aethiops ; Culture Media ; Guanidine/pharmacology ; Healthy Volunteers ; Humans ; RNA, Viral/genetics ; Reverse Transcriptase Polymerase Chain Reaction/methods ; SARS-CoV-2/drug effects ; SARS-CoV-2/genetics ; Saliva/drug effects ; Saliva/virology ; Specimen Handling/methods ; Vero Cells ; Virus Inactivation/drug effects
    Chemical Substances Culture Media ; RNA, Viral ; Guanidine (JU58VJ6Y3B)
    Language English
    Publishing date 2021-06-11
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0252687
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays

    Banik, Sukalyani / Saibire, Kaheerman / Suryavanshi, Shraddha / Johns, Glenn / Chakravorty, Soumitesh / Kwiatkawoski, Robert / Alland, David / Banada, Padmapriya P.

    medRxiv

    Abstract: ... inactivation medium for downstream RT-PCR testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 ... and processing of clinical samples for RT- PCR based SARS-CoV-2 detection. ... Background: Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and ...

    Abstract Background: Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings. Methods: We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream RT-PCR testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C. Results: SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log<sub>10</sub> PFU/ml within a minute of incubation. When saliva was diluted 1:1 in eNAT, no cytopathic effect (CPE) on Vero-E6 cell lines was observed, although SARS-CoV-2 RNA could be detected even after 30 min incubation and after two cell culture passages. A 1:2 (saliva:eNAT) dilution abrogated both  CPE and detectable viral RNA after as little as 5 min incubation in eNAT.  SARS-CoV-2 RNA from the virus spiked at 5X the limit of detection remained positive up to 7 days of incubation in all tested conditions. Conclusion: eNAT and similar guanidinium thiocyanate-based media may be of value for transport, preservation, and processing of clinical samples for RT- PCR based SARS-CoV-2 detection.
    Keywords covid19
    Language English
    Publishing date 2021-01-20
    Publisher Cold Spring Harbor Laboratory Press
    Document type Article ; Online
    DOI 10.1101/2021.01.15.21249891
    Database COVID19

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  4. Article ; Online: Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.

    Sukalyani Banik / Kaheerman Saibire / Shraddha Suryavanshi / Glenn Johns / Soumitesh Chakravorty / Robert Kwiatkowski / David Alland / Padmapriya P Banada

    PLoS ONE, Vol 16, Iss 6, p e

    2021  Volume 0252687

    Abstract: ... stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection. ... Background Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and ... present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 ...

    Abstract Background Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings. Methods We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C. Results SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log10 PFU/ml within a minute of incubation. When saliva was diluted 1:1 in eNAT, no cytopathic effect (CPE) on VeroE6 cells was observed, although SARS-CoV-2 RNA could be detected even after 30 min incubation and after two cell culture passages. A 1:2 (saliva:eNAT) dilution abrogated both CPE and detectable viral RNA after as little as 5 min incubation in eNAT. SARS-CoV-2 RNA from virus spiked at 5X the limit of detection remained positive up to 7 days of incubation in all tested conditions. Conclusion eNAT and similar guanidinium thiocyanate-based media may be of value for transport, stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection.
    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2021-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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