The size and culturability of patient-generated SARS-CoV-2 aerosol

Joshua L Santarpia; Vicki L Herrera; Danielle N Rivera; Shanna Ratnesar-Shumate; St Patrick Reid; Daniel N Ackerman; Paul W Denton; Jacob W S Martens; Ying Fang; Nicholas Conoan; Michael V Callahan; James V Lawler; David M Brett-Major; John J Lowe

doi:10.1038/s41370-021-00376-8

The size and culturability of patient-generated SARS-CoV-2 aerosol

Joshua L Santarpia, Vicki L Herrera, Danielle N Rivera, Shanna Ratnesar-Shumate, St Patrick Reid, Daniel N Ackerman, Paul W Denton, Jacob W S Martens, Ying Fang, Nicholas Conoan, Michael V Callahan, James V Lawler, David M Brett-Major, John J Lowe

Pathobiology

Research output: Contribution to journal › Article › peer-review

Abstract

BACKGROUND: Aerosol transmission of COVID-19 is the subject of ongoing policy debate. Characterizing aerosol produced by people with COVID-19 is critical to understanding the role of aerosols in transmission.

OBJECTIVE: We investigated the presence of virus in size-fractioned aerosols from six COVID-19 patients admitted into mixed acuity wards in April of 2020.

METHODS: Size-fractionated aerosol samples and aerosol size distributions were collected from COVID-19 positive patients. Aerosol samples were analyzed for viral RNA, positive samples were cultured in Vero E6 cells. Serial RT-PCR of cells indicated samples where viral replication was likely occurring. Viral presence was also investigated by western blot and transmission electron microscopy (TEM).

RESULTS: SARS-CoV-2 RNA was detected by rRT-PCR in all samples. Three samples confidently indicated the presence of viral replication, all of which were from collected sub-micron aerosol. Western blot indicated the presence of viral proteins in all but one of these samples, and intact virions were observed by TEM in one sample.

SIGNIFICANCE: Observations of viral replication in the culture of submicron aerosol samples provides additional evidence that airborne transmission of COVID-19 is possible. These results support the use of efficient respiratory protection in both healthcare and by the public to limit transmission.

Original language	English (US)
Pages (from-to)	706-711
Number of pages	6
Journal	Journal of Exposure Science and Environmental Epidemiology
Volume	32
Issue number	5
Early online date	Aug 18 2021
DOIs	https://doi.org/10.1038/s41370-021-00376-8
State	Published - Sep 2022

Keywords

SARS-CoV-2
human-generated aerosol
viral aerosol
aerosol transmission
COVID-19

ASJC Scopus subject areas

Public Health, Environmental and Occupational Health
Pollution
Epidemiology
Toxicology

Online availability

10.1038/s41370-021-00376-8License: CC BY

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Santarpia, J. L., Herrera, V. L., Rivera, D. N., Ratnesar-Shumate, S., Reid, S. P., Ackerman, D. N., Denton, P. W., Martens, J. W. S., Fang, Y., Conoan, N., Callahan, M. V., Lawler, J. V., Brett-Major, D. M., & Lowe, J. J. (2022). The size and culturability of patient-generated SARS-CoV-2 aerosol. Journal of Exposure Science and Environmental Epidemiology, 32(5), 706-711. https://doi.org/10.1038/s41370-021-00376-8

Santarpia, JL, Herrera, VL, Rivera, DN, Ratnesar-Shumate, S, Reid, SP, Ackerman, DN, Denton, PW, Martens, JWS, Fang, Y, Conoan, N, Callahan, MV, Lawler, JV, Brett-Major, DM & Lowe, JJ 2022, 'The size and culturability of patient-generated SARS-CoV-2 aerosol', Journal of Exposure Science and Environmental Epidemiology, vol. 32, no. 5, pp. 706-711. https://doi.org/10.1038/s41370-021-00376-8

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title = "The size and culturability of patient-generated SARS-CoV-2 aerosol",

abstract = "BACKGROUND: Aerosol transmission of COVID-19 is the subject of ongoing policy debate. Characterizing aerosol produced by people with COVID-19 is critical to understanding the role of aerosols in transmission.OBJECTIVE: We investigated the presence of virus in size-fractioned aerosols from six COVID-19 patients admitted into mixed acuity wards in April of 2020.METHODS: Size-fractionated aerosol samples and aerosol size distributions were collected from COVID-19 positive patients. Aerosol samples were analyzed for viral RNA, positive samples were cultured in Vero E6 cells. Serial RT-PCR of cells indicated samples where viral replication was likely occurring. Viral presence was also investigated by western blot and transmission electron microscopy (TEM).RESULTS: SARS-CoV-2 RNA was detected by rRT-PCR in all samples. Three samples confidently indicated the presence of viral replication, all of which were from collected sub-micron aerosol. Western blot indicated the presence of viral proteins in all but one of these samples, and intact virions were observed by TEM in one sample.SIGNIFICANCE: Observations of viral replication in the culture of submicron aerosol samples provides additional evidence that airborne transmission of COVID-19 is possible. These results support the use of efficient respiratory protection in both healthcare and by the public to limit transmission.",

keywords = "SARS-CoV-2, human-generated aerosol, viral aerosol, aerosol transmission, COVID-19",

author = "Santarpia, {Joshua L} and Herrera, {Vicki L} and Rivera, {Danielle N} and Shanna Ratnesar-Shumate and Reid, {St Patrick} and Ackerman, {Daniel N} and Denton, {Paul W} and Martens, {Jacob W S} and Ying Fang and Nicholas Conoan and Callahan, {Michael V} and Lawler, {James V} and Brett-Major, {David M} and Lowe, {John J}",

note = "This work was funded in part by a National Science Foundation COVID-19 RAPID Grant # 2032135. The authors would like to thank the Electron Microscopy Core Facility (EMCF) at the University of Nebraska Medical Center for technical assistance. The EMCF is supported by state funds from the Nebraska Research Initiative (NRI) and the University of Nebraska Foundation, and institutionally by the Office of the Vice Chancellor for Research. We also thank William Lindsley for the loan of the NIOSH BC251 samplers. Finally, we would also like to thank the nurses and doctors who helped facilitate access to the patient rooms for sampling, and to the patients themselves for allowing us access to their rooms during their stay at Nebraska Medicine.",

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doi = "10.1038/s41370-021-00376-8",

language = "English (US)",

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AU - Santarpia, Joshua L

AU - Herrera, Vicki L

AU - Rivera, Danielle N

AU - Ratnesar-Shumate, Shanna

AU - Reid, St Patrick

AU - Ackerman, Daniel N

AU - Denton, Paul W

AU - Martens, Jacob W S

AU - Fang, Ying

AU - Conoan, Nicholas

AU - Callahan, Michael V

AU - Lawler, James V

AU - Brett-Major, David M

AU - Lowe, John J

N1 - This work was funded in part by a National Science Foundation COVID-19 RAPID Grant # 2032135. The authors would like to thank the Electron Microscopy Core Facility (EMCF) at the University of Nebraska Medical Center for technical assistance. The EMCF is supported by state funds from the Nebraska Research Initiative (NRI) and the University of Nebraska Foundation, and institutionally by the Office of the Vice Chancellor for Research. We also thank William Lindsley for the loan of the NIOSH BC251 samplers. Finally, we would also like to thank the nurses and doctors who helped facilitate access to the patient rooms for sampling, and to the patients themselves for allowing us access to their rooms during their stay at Nebraska Medicine.

PY - 2022/9

Y1 - 2022/9

N2 - BACKGROUND: Aerosol transmission of COVID-19 is the subject of ongoing policy debate. Characterizing aerosol produced by people with COVID-19 is critical to understanding the role of aerosols in transmission.OBJECTIVE: We investigated the presence of virus in size-fractioned aerosols from six COVID-19 patients admitted into mixed acuity wards in April of 2020.METHODS: Size-fractionated aerosol samples and aerosol size distributions were collected from COVID-19 positive patients. Aerosol samples were analyzed for viral RNA, positive samples were cultured in Vero E6 cells. Serial RT-PCR of cells indicated samples where viral replication was likely occurring. Viral presence was also investigated by western blot and transmission electron microscopy (TEM).RESULTS: SARS-CoV-2 RNA was detected by rRT-PCR in all samples. Three samples confidently indicated the presence of viral replication, all of which were from collected sub-micron aerosol. Western blot indicated the presence of viral proteins in all but one of these samples, and intact virions were observed by TEM in one sample.SIGNIFICANCE: Observations of viral replication in the culture of submicron aerosol samples provides additional evidence that airborne transmission of COVID-19 is possible. These results support the use of efficient respiratory protection in both healthcare and by the public to limit transmission.

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KW - human-generated aerosol

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KW - aerosol transmission

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ER -

The size and culturability of patient-generated SARS-CoV-2 aerosol

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