Efficiency and sensitivity optimization of a protocol to quantify indoor airborne SARS-CoV-2 levels

Joan, Truyols-Vives; Kristiyan, Stiliyanov-Atanasov; Ernest, Sala-Llinàs; Toledo-Pons, Núria; Herme, G. Baldoví; Josep, Mercader-Barceló

doi:10.1016/j.jhin.2022.08.011

Cited by 5 publications

(8 citation statements)

References 35 publications

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“…This study is limited by units in which the usage of both quantitative (copies/μl) and qualitative (Ct value) detection approach with different primer probes for both approaches were compared. In addition to that differences in other PCR components for instance, the use of different reverse transcriptases, polymerases and number of cycles can also affects the detection of airborne SARS-CoV-2 ( 57 ). Secondly, there is also no consistence guidance on the better usage of either LVS or HVS in sampling strategy due to types of air sampler used.…”

Section: Discussionmentioning

confidence: 99%

Droplet digital PCR application for the detection of SARS-CoV-2 in air sample

Rashid,

Nazakat,

Muhamad Robat

et al. 2023

Front. Public Health

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Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) may transmit through airborne route particularly when the aerosol particles remain in enclosed spaces with inadequate ventilation. There has been no standard recommended method of determining the virus in air due to limitations in pre-analytical and technical aspects. Furthermore, the presence of low virus loads in air samples could result in false negatives. Our study aims to explore the feasibility of detecting SARS-CoV-2 ribonucleic acid (RNA) in air samples using droplet digital polymerase chain reaction (ddPCR). Active and passive air sampling was conducted between December 2021 and February 2022 with the presence of COVID-19 confirmed cases in two hospitals and a quarantine center in Klang Valley, Malaysia. SARS-CoV-2 RNA in air was detected and quantified using ddPCR and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The comparability of two different digital PCR platforms (QX200 and QIAcuity) to RT-PCR were also investigated. Additionally negative staining transmission electron microscopy was performed to visualize virus ultrastructure. Detection rates of SARS-CoV-2 in air samples using ddPCR were higher compared to RT-PCR, which were 15.2% (22/145) and 3.4% (5/145), respectively. The sensitivity and specificity of ddPCR was 100 and 87%, respectively. After excluding 17 negative samples (50%) by both QX200 and QIAcuity, 15% samples (5/34) were found to be positive both ddPCR and dPCR. There were 23.5% (8/34) samples that were detected positive by ddPCR but negative by dPCR. In contrast, there were 11.7% (4/34) samples that were detected positive by dPCR but negative by ddPCR. The SARS-CoV-2 detection method by ddPCR is precise and has a high sensitivity for viral RNA detection. It could provide advances in determining low viral titter in air samples to reduce false negative reports, which could complement detection by RT-PCR.

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Section: Discussionmentioning

confidence: 99%

Droplet digital PCR application for the detection of SARS-CoV-2 in air sample

Rashid,

Nazakat,

Muhamad Robat

et al. 2023

Front. Public Health

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“…Bioaerosol Collection. Liquid impingement was selected as the sampling method, following the example of Truyols-Vives et al [21,22], who were comparatively more efficient in collecting positive SARS-CoV-2 samples in the presence of COVID-19 patients than other researchers using different methods (e.g., Dumont-Leblond et al [14] and Stern et al [19]). In our study, airborne bioparticles were collected from the indoor air using a cyclonic impinger manufactured by Bertin Technologies (St-Berthely, France), Coriolis μ.…”

Section: Methodsmentioning

confidence: 99%

“…The inlet airflow rate ranges from 100 to 300 l min -1 on increments of 50 l min -1 , which represents an advantage compared with the impinger used by Truyols-Vives et al [21,22], that operated at the significantly lower flow rate of 12.5 l min -1 . This means that the Coriolis μ can sample a much larger total volume of air by applying the same sampling period as applied by Truyols-Vives et al [21,22]. The sampling period of the stand-alone Coriolis μ device can be set from 1 to 10 min, in increments of 1 min.…”

Section: Methodsmentioning

confidence: 99%

“…They attributed the good performance of their method to a combination of two main factors: the use of liquid impingement (SKC Biosampler®) to collect the virus (which helps to preserve the genetic material) and of digital droplet PCR (ddPCR) to analyze the samples (which is a newer technology that allows for lower detection limits compared to traditional qPCR). Later that same year, Truyols-Vives et al [22] optimized their sampling protocol even further by substituting their liquid sampling media by a mineral oil, with a much lower evaporation rate in the same sampling time. With this change, they were able to detect SARS-CoV-2 in 80% of the 15 samples collected in the presence of a COVID-19 patient.…”

Section: Introductionmentioning

confidence: 99%

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Development of a Bioaerosol Sampling Method for Airborne Pathogen Detection with Focus on SARS-CoV-2

Paralovo,

Vanden Driessche,

Cartuyvels

et al. 2024

Indoor Air

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As worldwide evidence shows that the predominant transmission route of SARS-CoV-2 and other respiratory pathogens is airborne, the need for suitable methods for the sampling of bioparticles directly from the air is more urgent than ever. The present paper describes the development of a method for the collection of biological aerosols, using a preexisting cyclonic impinger, the Coriolis μ, combined with a lysis buffer and subsequent qPCR analysis of the generated samples in lab. Four phases of method development are described: exploratory, validation, blank tests, and application. The application phase consisted of a field experiment in which the method was simultaneously applied at two daycare facilities. The method achieved a good level of accuracy and reliability in detecting different types of infectious agents in the air, with a global uncertainty of 19.6%. Furthermore, our method allows the simultaneous detection of 26 different respiratory pathogens in air samples, it is relatively simple, and the equipment is easy to use. Additionally, the time to collect a representative sample is short compared to other methods. The method does not cause significant disturbance to those present in the sampled rooms, and it is safe for operators and flexible, meaning it can be used in virtually any environment regardless of use, size, or occupancy. Further research is being developed to allow quantitative analysis of the collected samples and to test the methods’ ability to assess the viability of the microorganisms collected in the sample.

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“…Such studies provided insights into several transmissible places where healthcare workers (HCWs) are at risk of exposure to SARS‐CoV‐2, and have invariably shown the effectiveness of such techniques and their advantages in quantifying very low target copies and reduction of multiple processing. Further, enhancing sample collection through mineral oil, improvising viral RNA extraction protocols and cDNA synthesis efficiency can also be implemented 126 . Providing such additional improvements can be a practical advantage in understanding the viral dynamics in the air‐based transmission of the disease.…”

Section: Applications Of Digital Pcr In Sars‐cov‐2 Quantificationmentioning

confidence: 99%

SARS‐CoV‐2 in digital era: Diagnostic techniques and importance of nucleic acid quantification with digital PCRs

Aravind Kumar,

Aradhana,

Harleen

et al. 2023

Reviews in Medical Virology

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Studies related to clinical diagnosis and research of SARS‐CoV‐2 are important in the current pandemic era. Although molecular biology has emphasised the importance of qualitative analysis, quantitative analysis with nucleic acids in relation to SARS‐CoV‐2 needs to be clearly emphasised, which can provide perspective for viral dynamic studies of SARS‐CoV‐2. In this regard, the requirement and utilization of digital PCR in COVID‐19 research has substantially increased during the pandemic, necessitating the aggregation of its cardinal applications and future scopes. Hence, this meta‐review comprehensively addresses and emphasises the importance of nucleic acid quantification of SARS‐CoV‐2 RNA with digital PCR (dPCR). Various quantitative techniques of clinical significance like immunological, proteomic and nucleic acid‐based diagnosis and quantification, have been comparatively discussed. Furthermore, the core part of the article focusses on the working principle and advantages of digital PCR, along with its applications in COVID‐19 research. Several important applications like viral load quantitation, environmental surveillance and assay validation have been extensively investigated and discussed. Certain key future scopes of clinical importance, like mortality prediction, viral/variant‐symbiosis, and antiviral studies were also identified, suggesting several possible digital PCR applications in COVID‐19 research.

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Efficiency and sensitivity optimization of a protocol to quantify indoor airborne SARS-CoV-2 levels

Cited by 5 publications

References 35 publications

Droplet digital PCR application for the detection of SARS-CoV-2 in air sample

Droplet digital PCR application for the detection of SARS-CoV-2 in air sample

Development of a Bioaerosol Sampling Method for Airborne Pathogen Detection with Focus on SARS-CoV-2

SARS‐CoV‐2 in digital era: Diagnostic techniques and importance of nucleic acid quantification with digital PCRs

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