ABSTRACT
BACKGROUND: SARS-CoV-2 replicates efficiently in the upper airways of humans and produces high loads of virus RNA and, at least in the initial phase after infection, many infectious virus particles. Studying virus ultrastructure, such as particle integrity or presence of spike proteins, and effects on their host cells in patient samples is important to understand the pathogenicity of SARS-CoV-2. METHODS: Suspensions from swab samples with a high load of virus RNA (Ct < 20) were sedimented by desktop ultracentrifugation and prepared for thin section electron microscopy using a novel method which is described in detail. Embedding was performed in Epon or in LR White resin using standard or rapid protocols. Thin sections were examined using transmission electron microscopy. RESULTS: Virus particles could be regularly detected in the extracellular space, embedded in a background of heterogenous material (e.g. vesicles and needle-like crystals), and within ciliated cells. Morphology (i.e. shape, size, spike density) of virus particles in the swab samples was very similar to particle morphology in cell culture. However, in some of the samples the virus particles hardly revealed spikes. Infected ciliated cells occasionally showed replication organelles, such as double-membrane vesicles. The most common cells in all samples were keratinocytes from the mucosa and bacteria. CONCLUSIONS: The new method allows the ultrastructural visualization and analysis of coronavirus particles and of infected host cells from easy to collect naso/oropharyngeal patient swab samples.
Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Specimen Handling/methods , Microscopy, Electron, Transmission , RNAABSTRACT
BACKGROUND: Nasopharyngeal (NP) and oropharyngeal (OP) swab sampling for coronavirus disease 2019 (COVID-19) diagnosis may lead to release of particles of varying sizes and increase the exposure risk for health care workers (HCWs). However, there is limited evidence for effective methods to reduce occupational exposure from NP and OP swab sampling. This study aimed to reduce droplet-forming responses (DFRs) and the related exposure risk of NP and OP swab sampling by administering 10% lidocaine spray (LS) to the NP and OP areas prior to conducting swab tests. METHODS: This quasi-experimental study was conducted with 100 patients who presented to our tertiary care hospital with symptoms of COVID-19 between December 1 and 15, 2020. First, NP and OP swabbings were performed on each patient. Thereafter, LS was applied to the OP and NP regions, and the swab samples were taken once again. Frequency of DFRs and real-time polymerase chain reaction (RT-PCR) test results before and after LS application were recorded for comparison. In addition, the cycle threshold (Ct) was used as a proxy indicator for SARS-CoV-2 viral load in COVID-19 positive cases. FINDINGS: Significant differences in OP DFR frequencies before and after LS intervention were found (37% and 9%, respectively), as well as before and after NP DFR (31% and 18%, respectively). The mean Ct values for the positive samples did not differ before and after applying LS. CONCLUSION: Our results suggest that applying LS to the OP and NP area prior to swab testing reduces DFR frequencies without affecting (RT-PCR) test results for SARS-CoV-2 and may increase patient and practitioner comfort.
Subject(s)
COVID-19 , Health Personnel , Humans , Nasopharynx , SARS-CoV-2 , Specimen HandlingABSTRACT
In this study, 127 sputum/tracheal aspirate specimens were evaluated by a laboratory-developed real-time RT-PCR method and Fusion SARS-CoV-2 assay. These specimens were collected from the patients who have nasopharyngeal swab (NPS) samples being used for SARS-CoV-2 detection previously or simultaneously. The overall agreement was 96% between the lower respiratory tract (LRT) and NPS samples, suggesting that LRT specimens could be an option for patients who develop a productive cough or those receiving invasive mechanical ventilation.