Inactivating Zaire Ebolavirus in Whole-Blood Thin Smears Used for Malaria Diagnosis

Cutts, Todd; Cook, Bradley W. M.; Poliquin, P. Guillaume; Strong, James E.; Theriault, Steven

doi:10.1128/jcm.02960-15

Cited by 9 publications

(6 citation statements)

References 8 publications

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“…Therefore, UVC inactivation of clinical samples could lead to misleading biological interpretations, artificially skewing sample metabolites to a more oxidized profile (4951). However, highconcentration methanol ($80%) (59,60) and methanol/acetone mixtures (12,21) have previously been shown to successfully inactivate many viral-infected samples, including SARS-CoV-2 (1618). The extraction solvent we used is similar to that used in many metabolomic sample preparation techniques and was sufficient to inactivate virus while maintaining data integrity.…”

Section: Discussionmentioning

confidence: 99%

Inactivation of SARS-CoV-2 and COVID-19 Patient Samples for Contemporary Immunology and Metabolomics Studies

et al. 2022

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Due to the severity of COVID-19 disease, the U.S. Centers for Disease Control and Prevention and World Health Organization recommend that manipulation of active viral cultures of SARS-CoV-2 and respiratory secretions from COVID-19 patients be performed in biosafety level (BSL)3 laboratories. Therefore, it is imperative to develop viral inactivation procedures that permit samples to be transferred to lower containment levels (BSL2), while maintaining the fidelity of complex downstream assays to expedite the development of medical countermeasures. In this study, we demonstrate optimal conditions for complete viral inactivation following fixation of infected cells with commonly used reagents for flow cytometry, UVC inactivation in sera and respiratory secretions for protein and Ab detection, heat inactivation following cDNA amplification for droplet-based single-cell mRNA sequencing, and extraction with an organic solvent for metabolomic studies. Thus, we provide a suite of viral inactivation protocols for downstream contemporary assays that facilitate sample transfer to BSL2, providing a conceptual framework for rapid initiation of high-fidelity research as the COVID-19 pandemic continues. ImmunoHorizons, 2022, 6: 144-155.

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Section: Discussionmentioning

confidence: 99%

Inactivation of SARS-CoV-2 and COVID-19 Patient Samples for Contemporary Immunology and Metabolomics Studies

et al. 2022

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“…However, high concentration methanol (≥80%) (60,61) and methanol/acetone mixtures (13,22) have previously been shown to successfully inactivate many viral infected samples including SARS-CoV-2 (17)(18)(19). The extraction solvent we used is similar to that used in many metabolomic sample preparation techniques, and was sufficient to inactivate virus while maintaining data integrity.…”

Section: Discussionmentioning

confidence: 99%

Inactivation of SARS Coronavirus 2 and COVID-19 patient samples for contemporary immunology and metabolomics studies

Eddins

Bassit

Chandler

et al. 2021

Preprint

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In late 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged from Wuhan, China spurring the Coronavirus Disease-19 (COVID-19) pandemic that has resulted in over 219 million confirmed cases and nearly 4.6 million deaths worldwide. Intensive research efforts ensued to constrain SARS-CoV-2 and reduce COVID-19 disease burden. Due to the severity of this disease, the US Centers for Disease Control and Prevention (CDC) and World Health Organization (WHO) recommend that manipulation of active viral cultures of SARS-CoV-2 and respiratory secretions from COVID-19 patients be performed in biosafety level 3 (BSL3) containment laboratories. Therefore, it is imperative to develop viral inactivation procedures that permit samples to be transferred and manipulated at lower containment levels (i.e., BSL2), and maintain the fidelity of downstream assays to expedite the development of medical countermeasures (MCMs). We demonstrate optimal conditions for complete viral inactivation following fixation of infected cells with paraformaldehyde solution or other commonly-used branded reagents for flow cytometry, UVC inactivation in sera and respiratory secretions for protein and antibody detection assays, heat inactivation following cDNA amplification of single-cell emulsions for droplet-based single-cell mRNA sequencing applications, and extraction with an organic solvent for metabolomic studies. Thus, we provide a suite of protocols for viral inactivation of SARS-CoV-2 and COVID-19 patient samples for downstream contemporary immunology assays that facilitate sample transfer to BSL2, providing a conceptual framework for rapid initiation of high-fidelity research as the COVID-19 pandemic continues.

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“…Previous studies conducted by Cutts et al 15 at the Canadian Science Centre for Human and Animal Health laid the groundwork for a better understanding of the dynamics of EBOV inactivation in peripheral blood thin smears. Their work determined that absolute methanol completely inactivated EBOV-Makona after 15 minutes of exposure.…”

Section: Discussionmentioning

confidence: 99%

Methanol Fixation, but not Giemsa Staining, Inactivates Ebola and Lassa Viruses in Peripheral Blood Smears Made on Plastic Microscope Slides

Relich

Feldmann

Haddock

2020

The American Journal of Tropical Medicine and Hygiene

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Diseases caused by many highly pathogenic viruses, including Ebola virus (EBOV) and Lassa virus (LASV), present with nonspecific signs and symptoms that overlap with common tropical diseases such as malaria. Initial diagnostic tests performed on patients under investigation for viral hemorrhagic fevers routinely include analysis of peripheral blood smears to detect and quantify Plasmodium species. In light of recent and ongoing Ebola virus disease and Lassa fever epidemics, clinical laboratories around the world require protocols for dealing with highly infectious specimens from patients with suspected or confirmed high-consequence diseases. Few validated protocols for safe analysis of peripheral blood smears are available, revealing a need for further research. In this study, we evaluated the performance of two plastic microscope slide types that offer safe alternatives to glass slides, determined the temporal parameters required to inactivate EBOV and LASV in thin blood smears by methanol fixation, and assessed the virucidal activity of Giemsa stain. Both types of plastic microscope slides performed optimally; there were no significant differences in blood cell morphology or tinctorial properties nor were differences noted in Plasmodium ovale morphology or staining, when compared with glass slides. For both EBOV and LASV, viable viruses were not detected in thin blood smears following fixation in absolute methanol for at least 2 minutes. By contrast, viable EBOV and LASV were recovered from all Giemsa-stained thick blood smears.

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Inactivating Zaire Ebolavirus in Whole-Blood Thin Smears Used for Malaria Diagnosis

Cited by 9 publications

References 8 publications

Inactivation of SARS-CoV-2 and COVID-19 Patient Samples for Contemporary Immunology and Metabolomics Studies

Inactivation of SARS-CoV-2 and COVID-19 Patient Samples for Contemporary Immunology and Metabolomics Studies

Inactivation of SARS Coronavirus 2 and COVID-19 patient samples for contemporary immunology and metabolomics studies

Methanol Fixation, but not Giemsa Staining, Inactivates Ebola and Lassa Viruses in Peripheral Blood Smears Made on Plastic Microscope Slides

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