Gene

Coronavirus

2019-20 coronavirus outbreak

Persistence (discontinuity)

Sars virus

Engineering

Biopsy

Biology

Geotechnical engineering

Betacoronavirus

Virology

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)

Outbreak

Coronavirus disease 2019 (COVID-19)

Immunology

Lung transplantation

Genetics

Infectious disease (medical specialty)

Internal medicine

Medicine

Transmission (telecommunications)

Lung biopsy

Disease

Lung

Electrical engineering

Pathology

The Lancet Respiratory Medicine

On Dec 1, 2020, we reported a successful case of double-lung transplantation from a SARS-CoV-2 seropositive donor 105 days after the onset of mild COVID-19.1Ceulemans LJ Van Slambrouck J De Leyn P et al.Successful double-lung transplantation from a donor previously infected with SARS-CoV-2.Lancet Respir Med. 2021; 9: 315-318Summary Full Text Full Text PDF PubMed Scopus (42) Google Scholar Although repeated quantitative (q)RT-PCR analyses of donor nasopharyngeal swabs were negative, this technique detected RNA of the SARS-CoV-2 N gene (delta Ct 35) from a biopsy of the right lung taken during organ procurement. Viral culture of this biopsy was negative and donor-to-recipient transmission did not occur. Complementary orthogonal methods were needed to corroborate and interpret the qRT-PCR results. Therefore, we did ultrasensitive single-molecule fluorescence RNA in-situ hybridisation with RNAscope technology on formalin-fixed paraffin-embedded sections of the same lung biopsy (appendix p 1), and compared the results with those of a lung biopsy from a deceased patient with acute COVID-19 (figure A and B; appendix p 2). We stained 14 slides of the donor lung biopsy, each containing one 5 μm section, as follows: five slides with a probe for the N gene; five slides with a probe for the S gene; and four slides with probes for N and S. A probe for the basigin gene, which has been proposed to encode an alternative host recipient for SARS-CoV-2, served as a positive control on the ten slides stained for N or S only.2Wang K Chen W Zhang Z et al.CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells.Signal Transduct Target Ther. 2020; 5: 283Crossref PubMed Scopus (650) Google Scholar We identified characteristic RNAscope puncta in three out of nine slides for the N probe, and in six out of nine slides for the S probe (figure C and D). These puncta appeared to be located in clumps of sloughed-off material, and no cells or cell nuclei could be discerned in this debris-like tissue. To our knowledge, this is the first report of long-term (>100 days) persistence of SARS-CoV-2 RNA in lung tissue of an immunocompetent patient after convalescing from COVID-19. The debris-like tissue that contained SARS-CoV-2 RNA might be composed of degenerated endothelial cells that had detached from vessel walls, dysmorphic syncytial elements of pneumocytes, or dead neutrophilic plugs in the interstitium.3Bussani R Schneider E Zentilin L et al.Persistence of viral RNA, pneumocyte syncytia and thrombosis are hallmarks of advanced COVID-19 pathology.EBioMed. 2020; 61103104Summary Full Text Full Text PDF PubMed Scopus (234) Google Scholar, 4Schurink B Roos E Radonic T et al.Viral presence and immunopathology in patients with lethal COVID-19: a prospective autopsy cohort study.Lancet Microbe. 2020; 1: e290-e299Summary Full Text Full Text PDF PubMed Scopus (357) Google Scholar We speculate that this debris-like tissue might shield SARS-CoV-2 RNA from degradation. Data on sputum, nasopharyngeal swabs, and bronchoalveolar lavage fluid indicate that prolonged detection of SARS-CoV-2 RNA is rare and limited to a few weeks.5Wölfel VM Corman W Guggemos M et al.Virological assessment of hospitalized patients with COVID-2019.Nature. 2020; 581: 465-469Crossref PubMed Scopus (4681) Google Scholar By contrast, SARS-CoV-2 RNA persisted in the lung parenchyma for 105 days after the onset of a mild course of COVID-19. Nonetheless, at the time of writing, 11 months after transplantation, the recipient is in good health. Our data show that the persistence of SARS-CoV-2 RNA in this donor lung tissue has been inconsequential. LJC is supported by a Katholieke Universiteit Leuven University Chair funded by Medtronic and a post-doctoral grant from University Hospitals Leuven (KOOR-UZ Leuven). LVG is supported by a post-doctoral grant from University Hospitals Leuven (KOOR-UZ Leuven). AV is supported by a fundamental research grant from the Research Foundation Flanders (FWO). DVR is supported by a grant from the Broere Charitable Foundation. RV and JW are supported as senior clinical research fellows by the FWO Belgium. EW is supported by Stichting tegen Kanker (basic and clinical oncology research). All other authors declare no competing interests. Download .pdf (.34 MB) Help with pdf files Supplementary appendix Successful double-lung transplantation from a donor previously infected with SARS-CoV-2In late 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its resulting respiratory disease, COVID-19, emerged.1 As the pandemic continues, with more than 62 million people with documented SARS-CoV-2 infection, a growing number of potential organ donors will have been infected. Since SARS-CoV-2 resides primarily in the respiratory tract, lung transplantation has the highest concern in terms of donor-derived viral transmission and impaired graft quality. Therefore, graft acceptance for lung transplantation should carefully be balanced against the longer waiting time for the recipient. Full-Text PDF

Persistence of SARS-CoV-2 RNA in lung tissue after mild COVID-19