Monitoring COVID-19 Transmission Risks by Quantitative Real-Time PCR Tracing of Droplets in Hospital and Living Environments.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) environmental contamination occurs through droplets and biological fluids released in the surroundings from patients or asymptomatic carriers. Surfaces and objects contaminated by saliva or nose secretions represent a risk for indirect transmission of coronavirus disease 2019 (COVID-19). We assayed surfaces from hospital and living spaces to identify the presence of viral RNA and the spread of fomites in the environment. Anthropic contamination by droplets and biological fluids was monitored by detecting the microbiota signature using multiplex quantitative real-time PCR (qPCR) on selected species and massive sequencing on 16S amplicons. A total of 92 samples (flocked swabs) were collected from critical areas during the pandemic, including indoor (three hospitals and three public buildings) and outdoor surfaces exposed to anthropic contamination (handles and handrails, playgrounds). Traces of biological fluids were frequently detected in spaces open to the public and on objects that are touched with the hands (>80%). However, viral RNA was not detected in hospital wards or other indoor and outdoor surfaces either in the air system of a COVID hospital but only in the surroundings of an infected patient, in consistent association with droplet traces and fomites. Handled objects accumulated the highest level of multiple contaminations by saliva, nose secretions, and fecal traces, further supporting the priority role of handwashing in prevention. In conclusion, anthropic contamination by droplets and biological fluids is widespread in spaces open to the public and can be traced by qPCR. Monitoring fomites can support evaluation of indirect transmission risks for coronavirus or other flu-like viruses in the environment.IMPORTANCE Several studies have evaluated the presence of SARS-CoV-2 in the environment. Saliva and nasopharyngeal droplets can land on objects and surfaces, creating fomites. A suitable indicator would allow the detection of droplets or biofluids carrying the virus. Therefore, we searched for viral RNA and droplets and fomites on at risk surfaces. We monitored by qPCR or next generation sequencing (NGS) droplets through their microbiota. Although the study was performed during the pandemic, SARS-CoV-2 was not significantly found on surfaces, with the only exception of environmental areas near infectious patients. Conversely, anthropic contamination was frequent, suggesting a role for biofluids as putative markers of indirect transmission and risk assessment. Moreover, all SARS-CoV-2-contaminated surfaces showed droplets' microbiota. Fomite monitoring by qPCR may have an impact on public health strategies, supporting prevention of indirect transmission similarly to what is done for other communicable diseases (e.g., influenza and influenza-like infections).

Piana A ，Colucci ME ，Valeriani F ，Marcolongo A ，Sotgiu G ，Pasquarella C ，Margarucci LM ，Petrucca A ，Gianfranceschi G ，Babudieri S ，Vitali P ，D'Ermo G ，Bizzarro A ，De Maio F ，Vitali M ，Azara A ，Romano F ，Simmaco M ，Romano Spica V ... -  《mSphere》

Increasing Temperature and Relative Humidity Accelerates Inactivation of SARS-CoV-2 on Surfaces.

Coronavirus disease 2019 (COVID-19) was first identified in China in late 2019 and is caused by newly identified severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Previous studies had reported the stability of SARS-CoV-2 in cell culture media and deposited onto surfaces under a limited set of environmental conditions. Here, we broadly investigated the effects of relative humidity, temperature, and droplet size on the stability of SARS-CoV-2 in a simulated clinically relevant matrix dried on nonporous surfaces. The results show that SARS-CoV-2 decayed more rapidly when either humidity or temperature was increased but that droplet volume (1 to 50 μl) and surface type (stainless steel, plastic, or nitrile glove) did not significantly impact decay rate. At room temperature (24°C), virus half-life ranged from 6.3 to 18.6 h depending on the relative humidity but was reduced to 1.0 to 8.9 h when the temperature was increased to 35°C. These findings suggest that a potential for fomite transmission may persist for hours to days in indoor environments and have implications for assessment of the risk posed by surface contamination in indoor environments.IMPORTANCE Mitigating the transmission of SARS-CoV-2 in clinical settings and public spaces is critically important to reduce the number of COVID-19 cases while effective vaccines and therapeutics are under development. SARS-CoV-2 transmission is thought to primarily occur through direct person-to-person transfer of infectious respiratory droplets or through aerosol-generating medical procedures. However, contact with contaminated surfaces may also play a significant role. In this context, understanding the factors contributing to SARS-CoV-2 persistence on surfaces will enable a more accurate estimation of the risk of contact transmission and inform mitigation strategies. To this end, we have developed a simple mathematical model that can be used to estimate virus decay on nonporous surfaces under a range of conditions and which may be utilized operationally to identify indoor environments in which the virus is most persistent.

Biryukov J ，Boydston JA ，Dunning RA ，Yeager JJ ，Wood S ，Reese AL ，Ferris A ，Miller D ，Weaver W ，Zeitouni NE ，Phillips A ，Freeburger D ，Hooper I ，Ratnesar-Shumate S ，Yolitz J ，Krause M ，Williams G ，Dawson DG ，Herzog A ，Dabisch P ，Wahl V ，Hevey MC ，Altamura LA ... -  《mSphere》

Detection and infectivity potential of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) environmental contamination in isolation units and quarantine facilities.

Environmental surfaces have been suggested as likely contributors in the transmission of COVID-19. This study assessed the infectivity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) contaminating surfaces and objects in two hospital isolation units and a quarantine hotel. SARS-CoV-2 virus stability and infectivity on non-porous surfaces was tested under controlled laboratory conditions. Surface and air sampling were conducted at two COVID-19 isolation units and in a quarantine hotel. Viral RNA was detected by RT-PCR and infectivity was assessed by VERO E6 CPE test. In laboratory-controlled conditions, SARS-CoV-2 gradually lost its infectivity completely by day 4 at ambient temperature, and the decay rate of viral viability on surfaces directly correlated with increase in temperature. Viral RNA was detected in 29/55 surface samples (52.7%) and 16/42 surface samples (38%) from the surroundings of symptomatic COVID-19 patients in isolation units of two hospitals and in a quarantine hotel for asymptomatic and very mild COVID-19 patients. None of the surface and air samples from the three sites (0/97) were found to contain infectious titres of SARS-Cov-2 on tissue culture assay. Despite prolonged viability of SARS-CoV-2 under laboratory-controlled conditions, uncultivable viral contamination of inanimate surfaces might suggest low feasibility for indirect fomite transmission.

Ben-Shmuel A ，Brosh-Nissimov T ，Glinert I ，Bar-David E ，Sittner A ，Poni R ，Cohen R ，Achdout H ，Tamir H ，Yahalom-Ronen Y ，Politi B ，Melamed S ，Vitner E ，Cherry L ，Israeli O ，Beth-Din A ，Paran N ，Israely T ，Yitzhaki S ，Levy H ，Weiss S ... -  《-》

COVID-19 global pandemic planning: Presence of SARS-CoV-2 fomites in a university hospital setting.

Bartlett C ，Langsjoen J ，Cheng Q ，Yingling AV ，Weiss M ，Bradfute S ，Perkins DJ ，Hurwitz I ... -  《-》

Environmental Contamination With SARS-CoV-2 in a Hospital Setting.

Background The coronavirus disease 2019 (COVID-19) pandemic is a global concern and has changed the way we practice medicine in acute hospital settings. This is particularly true with regard to patient triage, patient risk assessment, use of personal protective equipment, and environmental disinfection. Transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is primarily through inhalation of respiratory droplets generated through talking, coughing, or sneezing. There is, however, a potential risk that respiratory droplets settling on inanimate surfaces and objects in the hospital environment could provide a reservoir for nosocomial infections in patients and pose a healthcare risk to medical staff. Indeed, there have been previous reports of healthcare-associated outbreaks in hospitals. Several authors have argued that the risk of transmission via fomites may be insignificant. It is, however, not clear what proportion of SARS-CoV-2 infections are attributable to direct contact with fomites; a few reports have indicated possible transmission via this route. Environmental contamination with SARS-CoV-2 in healthcare institutions has been shown to vary according to the function or service provided by a unit or department. Information that identifies hospital areas that have a propensity for higher environmental burden may improve the practice of infection control and environmental cleaning and decontamination in healthcare institutions. This study aimed to investigate environmental SARS-CoV-2 contamination in the clinical areas of patients with COVID-19 infection. Methodology We conducted a cross-sectional study performing swabbing of frequently touched surfaces, equipment, and ventilation ducts in five specific clinical areas of Peterborough City Hospital which is part of the North West Anglia NHS Foundation Trust. The five clinical areas that were chosen for swabbing were the Emergency Department (ED), Intensive Care Unit (ICU), Isolation Ward, Respiratory Ward, and a Gastroenterology Ward that was serving as a receiving ward at the height of the second COVID-19 infection wave in the United Kingdom. Surfaces to be swabbed were divided into the patient zone, doctor zone, and nursing zone. Swabs from the chosen surfaces were collected on two consecutive days. A total of 158 surface swabs were collected during the second wave of the COVID-19 pandemic. SARS-CoV-2 RNA was detected by reverse transcription polymerase chain reaction. Results The most contaminated clinical areas were the three receiving wards where 12% (11/96) of the swabs were positive. Inside the patient rooms, these surfaces included bed rails and controls, bedside tables, television screens, remote control units, and the room ventilation system. Outside the patient room, these surfaces included mobile computers and computer desk surfaces in the doctors' offices. All swabs taken from the ED and ICU were found to be negative. Conclusions Our study confirms the potential infection risks posed by environmental contamination with the SARS-CoV-2 virus. This highlights the importance of adequate environmental cleaning for proper infection control and prevention in healthcare settings.

Kumar S ，Bukhari I ，Afzal Z ，Tucker S ，Lucas-Evans R ，Dayala A ，Mlangeni D ... -  《Cureus》