Variable levels of spike and ORF1ab RNA in post-mortem lung samples of SARS-CoV-2-positive subjects: comparison between ISH and RT-PCR
| Autor: | Federica Zito Marino, Tiziana De Cristofaro, Massimo Varriale, Giuseppa Zannini, Andrea Ronchi, Elvira La Mantia, Carlo Pietro Campobasso, Francesco De Micco, Pasquale Mascolo, Maurizio Municinò, Emilia Municinò, Francesco Vestini, Omero Pinto, Marta Moccia, Noè De Stefano, Oscar Nappi, Carmen Sementa, Giovanni Zotti, Lamberto Pianese, Carmela Giordano, Renato Franco |
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| Přispěvatelé: | Zito Marino, F., De Cristofaro, T., Varriale, M., Zannini, G., Ronchi, A., La Mantia, E., Campobasso, C. P., De Micco, F., Mascolo, P., Municino, M., Municino, E., Vestini, F., Pinto, O., Moccia, M., De Stefano, N., Nappi, O., Sementa, C., Zotti, G., Pianese, L., Giordano, C., Franco, R. |
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
Open reading frame (ORF1ab)
Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 In situ hybridization (ISH) Post-mortem lung samples Post-mortem lung sample COVID-19 virus diseases Cell Biology General Medicine Spike(S) Sensitivity and Specificity Pathology and Forensic Medicine Reverse transcription polymerase chain reaction (RT-PCR) Humans RNA Viral Original Article Molecular Biology Lung In Situ Hybridization |
| Zdroj: | Virchows Archiv |
| Popis: | Post-mortem examination plays a pivotal role in understanding the pathobiology of the SARS-CoV-2; thus, the optimization of virus detection on the post-mortem formalin-fixed paraffin-embedded (FFPE) tissue is needed. Different techniques are available for the identification of the SARS-CoV-2, including reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), in situ hybridization (ISH), and electron microscopy. The main goal of this study is to compare ISH versus RT-PCR to detect SARS-CoV-2 on post-mortem lung samples of positive deceased subjects. A total of 27 samples were analyzed by RT-PCR targeting different viral RNA sequences of SARS-CoV-2, including envelope (E), nucleocapsid (N), spike (S), and open reading frame (ORF1ab) genes and ISH targeting S and Orf1ab. All 27 cases showed the N gene amplification, 22 out of 27 the E gene amplification, 26 out of 27 the S gene amplification, and only 6 the ORF1ab gene amplification. The S ISH was positive only in 12 out of 26 cases positive by RT-PCR. The S ISH positive cases with strong and diffuse staining showed a correlation with low values of the number of the amplification cycles by S RT-PCR suggesting that ISH is a sensitive assay mainly in cases carrying high levels of S RNA. In conclusion, our findings demonstrated that ISH assay has lower sensitivity to detect SARS-CoV-2 in FFPE compared to RT-PCR; however, it is able to localize the virus in the cellular context since it preserves the morphology. Supplementary Information The online version contains supplementary material available at 10.1007/s00428-021-03262-8. |
| Databáze: | OpenAIRE |
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