Post-Mortem RT-PCR Assay for SARS-CoV-2 RNA in COVID-19 Patients' Corneal Epithelium, Conjunctival and Nasopharyngeal Swabs.
20 Sep 2021-Journal of Clinical Medicine (Multidisciplinary Digital Publishing Institute)-Vol. 10, Iss: 18, pp 4256
TL;DR: In this paper, the authors investigated the post-mortem presence of SARS-CoV-2 RNA in corneal epithelium gathered by patients with an ante-mortem confirmed diagnosis of Coronavirus disease-19 (COVID-19).
Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease has been described to possibly be associated with ocular surface disturbances. However, whether the virus could invade ocular tissues still remains elusive. In the present study, we tried to investigate the post-mortem presence of SARS-CoV-2 RNA in corneal epithelium gathered by patients with an ante-mortem confirmed diagnosis of Coronavirus disease-19 (COVID-19). Cadavers with an ante-mortem confirmed diagnosis of moderate to severe COVID-19 were examined. Clinical and demographic features were retrieved from hospital patients' notes. For each cadaver, corneal scrapings, conjunctival swabs (CS) and nasopharyngeal swabs (NPS) were collected to perform real-time reverse transcriptase polymerase chain reaction ((RT)-PCR) for SARS-CoV-2. Fourteen consecutive cadavers with an ante-mortem confirmed diagnosis of moderate to severe COVID-19 were examined. The last NPS performed ante-mortem confirmed SARS-CoV-2 infection in 12/14 (85.7%) patients. The mean death-to-swab time (DtS) was 3.15 ± 0.5 (2.10-5.1) h. The post-mortem NPS and CS found positive for SARS-CoV-2 RNA were 9/14 (64.3%) and 3/28 (10.7%), respectively. None of the corneal epithelium scrapes tested positive to RT-PCR for SARS-CoV-2 RNA. These data promote the SARS-CoV-2 as not able to contaminate the post-mortem corneal epithelium, while it can persist in different other structures of the ocular surface (i.e., the conjunctiva). It is reasonable to assume that such a contamination can occur ante-mortem too.
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15 Oct 2021
TL;DR: In this article, the authors investigated the transmission risks of SARS-CoV-2 infection for subjects involved in handling, transporting, and examining deceased persons with known or suspected COVID-19 positivity at the time of death.
Abstract: To date, little is known regarding the transmission risks of SARS-CoV-2 infection for subjects involved in handling, transporting, and examining deceased persons with known or suspected COVID-19 positivity at the time of death. This experimental study aims to define if and/or how long SARS-CoV-2 persists with replication capacity in the tissues of individuals who died with/from COVID-19, thereby generating infectious hazards. Sixteen patients who died with/from COVID-19 who underwent autopsy between April 2020 and April 2021 were included in this study. Based on PMI, all samples were subdivided into two groups: ‘short PMI’ group (eight subjects who were autopsied between 12 to 72 h after death); ‘long PMI’ (eight subjects who were autopsied between 24 to 78 days after death). All patients tested positive for RT-PCR at nasopharyngeal swab both before death and on samples collected during post-mortem investigation. Moreover, a lung specimen was collected and frozen at −80 °C in order to perform viral culture. The result was defined based on the cytopathic effect (subjective reading) combined with the positivity of the RT-PCR test (objective reading) in the supernatant. Only in one sample (PMI 12 h), virus vitality was demonstrated. This study, supported by a literature review, suggests that the risk of cadaveric infection in cases of a person who died from/with COVID-19 is extremely low in the first hours after death, becoming null after 12 h after death, confirming the World Health Organization (WHO) assumed in March 2020 and suggesting that the corpse of a subject who died from/with COVID-19 should be generally considered not infectious.
6 citations
TL;DR: The increase of EKs, despite the pandemic, is a sign that the advance in corneal transplantation has gone ahead and it continues to evolve, and the Italian government adopted severe measures to limit viral transmission.
Abstract: Purpose To analyze the impact of COVID-19 on Italian corneal transplantation from March-2020 to February 2021 compared to the same timeframe of the 2 previous years, in order to identify potential consequences of a global pandemic on corneal procurement and transplantation services during this time. Methods This national, multicentric, retrospective cohort study evaluated data collected from 12 (100%) Italian eye banks from March 2020 to February 2021 (Group A). The number of tissues collected, distributed and discarded were compared with the same time-frame of the 2 previous years: 2019 and 2018 (group B and C, respectively). The different type of transplants performed were reported. Data were analyzed using a non-parametric Friedman test. Results Corneal procurement and the percentage of distributed tissues reduced in 2020 by more than 30 and 15%, respectively, compared to the 2 previous years. During the pandemic corneal transplant surgery showed only a modest drop: the number of the penetrating keratoplasties (PKs) and the anterior lamellar keratoplasties (ALKs) decreased by about 30 and 20% in comparison with groups B and C, respectively; between the Endothelial Keratoplasties (EKs), the Descemet membrane endothelial keratoplasty (DMEK) increased slightly from March 2020 to February 2021. Conclusions Italy was one of the first countries most affected by the outbreak of COVID-19, and the Italian government adopted severe measures to limit viral transmission. The pandemic generated several implications in corneal transplant activity during the first lockdown. Then an efficacious reaction with constant, vigorous work led to a resumption of transplant surgery to a near-normal standard. The increase of EKs, despite the pandemic, is a sign that the advance in corneal transplantation has gone ahead and it continues to evolve.
4 citations
28 Apr 2022
TL;DR: In this article , a retrospective review of electronic medical records of the ophthalmological departments of the University of Cagliari (SGD) and University Magna Græcia of Catanzaro (UMG), from 16 March 2020 to 14 March 2021 (52 weeks), were compared with data from the corresponding period of the previous year.
Abstract: During the COVID-19 era, several restrictions on surgery have been imposed to reduce the infectious risk among patients and staff and further preserve the availability of critical care resources. The aim of the study was to assess their impact on the ophthalmological practice and its medico-legal implications. A retrospective review of electronic medical records of the ophthalmological departments of the University of Cagliari (SGD) and University Magna Græcia of Catanzaro (UMG), from 16 March 2020 to 14 March 2021 (52 weeks), were compared with data from the corresponding period of the previous year. Weekly data on the number and type of diagnoses and procedures performed were collected and analysed in relation to the weekly average of the total number of COVID-19 patients in intensive care units (ICUs) and inpatients in Sardinia and Calabria. Results showed a significant decrease in cataract surgery operations by 47% and 31%, respectively, in the SGD and UMG (p < 0.05) during the second semester of the year; this drop occurred at the same time as the increase in COVID-19 patients in ICU and those hospitalised in both regions. Additionally, anterior segment surgery decreased at the UMG by 30% (p < 0.05). Vitreoretinal surgery decreased by 27% at the SGD, differently increased amount 31.5% at UMG (p < 0.05). The pandemic had a dramatic impact on elective surgery in ophthalmology, quantifying the backlog is the first step in order to understanding the measures to be taken in near future.
1 citations
TL;DR: In this article , the presence of viruses in postmortem nasopharyngeal swabs and ocular tissues of patients with coronavirus disease 19 (COVID-19) confirmed through an antemortem reverse transcription polymerase chain reaction (RT-PCR) test.
Abstract: The purpose of this study was to investigate the presence of viruses in postmortem nasopharyngeal swabs and ocular tissues of patients with coronavirus disease 19 (COVID-19) confirmed through an antemortem reverse transcription polymerase chain reaction (RT-PCR) test.We recorded demographic data, comorbidities, treatments, and vaccination status of the patients diagnosed with COVID-19 and monitored in the intensive care unit. Nasopharyngeal swab samples were collected from the patients within 2 hours postmortem followed by swabs from both eyes, lower and upper conjunctival sacs, corneal epithelial layer, and samples from anterior chamber fluid. The geneMAP 2019-nCoV severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) detection kit was used to detect the samples showing the presence of SARS-CoV-2.Among the 47 patients (mean age 74 years, range 39-92) included in the study, 29 were men and 18 were women. The nasopharyngeal swab samples of 21 patients were positive for the RT-PCR test for SARS-CoV-2, whereas at least one of the ocular tissue samples in 6 of those patients were positive for the RT-PCR test for SARS-CoV-2. However, all ocular swab samples were tested negative for SARS-CoV-2 in the 26 patients with negative nasopharyngeal swab samples. The mean duration from COVID-19 diagnosis to mortality was 10 days (range 2-27 days) in patients with positive postmortem nasopharyngeal swab samples for SARS-CoV-2, compared with an average of 21 days (1-80 days) in patients who tested negative. This difference was statistically significant (P = 0.01).Cadavers with negative postmortem nasopharyngeal swab samples for SARS-CoV-2 and those with the time from diagnosis to death of >3 weeks may be considered as candidates for corneal donation.
1 citations
TL;DR: The data suggest that corneal grafts may not be permissive for replication of SARS-CoV-2 and demonstrates that PCR testing of culture media cannot safely exclude that tissue has been exposed to SARS, as well as demonstrating the difficulty to differentiate between virus adherence and virus replication by PCR testing in Sars-Cov-2 exposed tissue.
Abstract: Concerns of possible transmission of SARS-CoV-2 from donors to patients by corneal transplantation have caused a decline in corneal transplantations. Graft culture media are routinely tested for infectious risks, but it is unclear whether this constitutes a viable means to avoid transmitting SARS-CoV-2 via keratoplasty. We found that SARS-CoV-2 RNA was not present in the medium after seven days of organ culture of corneas from donors (n = 4), who were SARS-CoV-2-positive upon tissue procurement. These medium samples showed no presence of viral RNA. To pursue this question under controlled conditions and further exclude the possibility of productive infection in corneal grafts, we inoculated corneoscleral discs from healthy donors (n = 8) with infectious SARS-CoV-2 and performed PCR testing of the culture medium at various time points. After seven days of culture, we also tested for SARS-CoV-2 RNA within the inoculated corneal tissue. The medium from tissue samples inoculated with SARS-CoV-2 showed no increase in viral RNA, which may indicate lack of viral replication in these corneal grafts. SARS-CoV-2-RNA was, however, detected on or in corneal tissue seven days after inoculation. Our data suggest that corneal grafts may not be permissive for replication of SARS-CoV-2 and demonstrates that PCR testing of culture media cannot safely exclude that tissue has been exposed to SARS-CoV-2. It also demonstrates the difficulty to differentiate between virus adherence and virus replication by PCR testing in SARS-CoV-2 exposed tissue.
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