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Infectious SARS-CoV-2 isolated from patient room air samples frozen for more than a year

A) ward layout for COVID-19 patients depicting sampling equipment location and distance to head SKC 37mm closed Face boxes effectively cut off aerosol sampling rather than liquid droplets. They are used downwards to prevent accidental contamination of droplets. 110A Series Liquid Point Samplers point samplers collect small size aerosols from 5nm to 10μm. B) by RT-qPCR Quantification of SARS-CoV-2 ORF1β RNA (Ct, purple bar, left y-axis) and extrapolation of genome equivalents/m
3 30 Air in collected air samples (blue bars, right y-axis). Red rectangles indicate four samples analyzed in cell culture for the presence of infectious virus. Nasopharyngeal swabs (NPS) obtained from each sample Ct CE) to assess cytopathic effect (CPE), expression of spike (S) and nucleocapsid (N), and de novo virion production in VERO E6 cells infected with 150 pfu of replication and β-propiolactone (BPL) Test experiment – inactivated SARS CoV-2. Parameters were evaluated on day 0 (D0) and day 3 (D3) of infection. C) CPE was detected by brightfield microscopy. D) Using polyclonal anti-SARS Related coronavirus 2 spike glycoprotein (BEI resources, cat#NR-52947) and SARS nucleocapsid protein antibody (Novus Biologicals, cat#) Western blot detection of SARS-CoV-2 S and N protein cells expressing NB100- 56576). E) Quantification of nascent virion production in supernatants of infected cells by median tissue culture infectious dose (TCID50) calculation. F–I) Analysis of air samples collected in COVID-19 patient wards. F) For Experimental protocol for culturing SARS-CoV-2 from air samples. Air samples were used as inoculum for VERO E6 cells. After 3 days of infection, the supernatant was collected and used to inoculate fresh VERO E6 cells. Use 150 pfu of SARS-CoV2/SB2 Isolates were subjected to control infection. Cellular expression of CPE (G), S and N proteins (H) and production of de novo virions (I) were monitored. ND: Not detectable. NA: Not available. Image credit:
Clinical Microbiology and Infection

(2023). DOI: 10.1016/j.cmi.2023.03.019

Quebec scientists have successfully isolated an infectious particle of the SARS-CoV-2 virus A new study shows that air samples from COVID-19 patient wards were collected and frozen for more than a year.

The study was carried out by a team led by Nathalie Grandvaux, researcher at the CHUM Research Center (CRCHUM) and professors at the University of Montreal, in collaboration with the teams of Caroline Duchaine (University Laval) and Yves Longtin (McGill University).

Published in Clinical Microbiology and Infection , the study provides insights into an area of ​​science that has been little explored since the pandemic began.

“In our study, we showed through an experimental model that it is possible Isolate and grow infectious virus from air samples,” says Nathalie Grandvaux. “This is possible even if samples were collected more than a year ago, then frozen and stored prior to cell culture.”

If public health authorities, including the World Health Organization, have been slow to recognize airborne viruses, it is partly because of limited scientific evidence for the presence of infectious viral particles in aerosols.

“These studies are hard to do,” says Nathalie Grandvaux, “because you need To maintain virus infectivity during collection, use proper cell culture techniques and access to a level 3 containment laboratory such as that of CRCHUM.”

unique method Audray Fortin, a researcher in the group of Nathalie Grandvaux and first author of the study, developed a unique cell culture method to amplify viruses collected by Canadian bioaerosol expert Caroline Duchaine and her team.

That was another feat of science that required a sampler – placed inside A range of collection equipment for COVID-19 patient rooms – to keep the virus infectious and protect it during storage.

During the study period, a total of 30 samples of COVID-19 and then stored frozen in the biobank for 14 months.

Using aerosol samples from a patient room, team of scientists are able to identify replicating virus the presence of particles.

“Thanks to our method it was possible to retrospectively assess the infectivity The presence of the virus SARS-CoV-2 in samples collected during different waves of the pandemic,” says Nathalie Grandvaux. “These data will help us better understand the importance of airborne transmission of the virus and implement adaptive prevention strategies.”

Be better prepared for the next pandemic

findings could be used to prepare for the next pandemic, whether it be SARS-CoV-2 or another respiratory virus, she added.

“Our study should raise awareness of airborne infectious viruses .It justifies targeting airborne transmission in individual and collective protective measures, including improving indoor air quality.”

The collection and culture method could also be adapted to closed environments outside of hospitals, such as schools, to test air quality and assess the effectiveness of protective measures against airborne viruses.

Further information:
Audray Fortin et al., Detection of liveness in retrospective analysis of aerosol samples collected from hospital rooms SARS-CoV-2 COVID-19 Patients, Clinical Microbiology and Infection (2023). DOI: 10.1016/j.cmi.2023.03.019

Courtesy of Montreal University Hospital Research center

Citation : Isolation from a hospital room air sample frozen for more than a year (April 19, 2023) Infectious SARS-CoV-2, Retrieved May 3, 2023 from

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