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.
Highlights
Since December 2019, the emergence of the novel beta coronavirus SARS-CoV-2 has been causing a devastating global pandemic that far surpasses its predecessors, SARS-CoV and MERS-CoV, in case numbers [1,2]
It has been established that SARS-CoV-2 enters target cells via interaction of its spike glycoprotein and the angiotensin-converting enzyme 2 (ACE2) receptor
We investigated whether SARS-CoV-2 RNA could be detected in the culture supernatant of corneal grafts from deceased patients who were confirmed SARS-CoV-2 positive by PCR testing of nasopharyngeal swabs, obtained at the time of tissue procurement
Summary
Since December 2019, the emergence of the novel beta coronavirus SARS-CoV-2 has been causing a devastating global pandemic that far surpasses its predecessors, SARS-CoV and MERS-CoV, in case numbers [1,2]. It has been established that SARS-CoV-2 enters target cells via interaction of its spike glycoprotein and the angiotensin-converting enzyme 2. It requires proteolytic cleavage of the viral spike protein by host proteases, with type II transmembrane serine protease (TMPRSS2) being the main interaction partner [6]. Results from extensive RNA sequencing approaches and immunohistochemical analysis demonstrate the expression of both ACE2 receptor and TMPRSS2 on corneal and conjunctival epithelial cells [7,8], suggesting that the cornea might be targeted by SARS-CoV-2.
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