scholarly journals Multisystemic Cellular Tropism of SARS-CoV-2 in Autopsies of COVID-19 Patients

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1900
Author(s):  
Dickson W. L. Wong ◽  
Barbara M. Klinkhammer ◽  
Sonja Djudjaj ◽  
Sophia Villwock ◽  
M. Cherelle Timm ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Viral Rna ◽  
Transmission Electron ◽  
Formalin Fixed Paraffin ◽  
Organ Systems ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues ◽  
Rna In Situ Hybridization

Multiorgan tropism of SARS-CoV-2 has previously been shown for several major organs. We have comprehensively analyzed 25 different formalin-fixed paraffin-embedded (FFPE) tissues/organs from autopsies of fatal COVID-19 cases (n = 8), using histopathological assessment, detection of SARS-CoV-2 RNA using polymerase chain reaction and RNA in situ hybridization, viral protein using immunohistochemistry, and virus particles using transmission electron microscopy. SARS-CoV-2 RNA was mainly localized in epithelial cells across all organs. Next to lung, trachea, kidney, heart, or liver, viral RNA was also found in tonsils, salivary glands, oropharynx, thyroid, adrenal gland, testicles, prostate, ovaries, small bowel, lymph nodes, skin and skeletal muscle. Viral RNA was predominantly found in cells expressing ACE2, TMPRSS2, or both. The SARS-CoV-2 replicating RNA was also detected in these organs. Immunohistochemistry and electron microscopy were not suitable for reliable and specific SARS-CoV-2 detection in autopsies. These findings were validated using in situ hybridization on external COVID-19 autopsy samples (n = 9). Apart from the lung, correlation of viral detection and histopathological assessment did not reveal any specific alterations that could be attributed to SARS-CoV-2. In summary, SARS-CoV-2 and its replication could be observed across all organ systems, which co-localizes with ACE2 and TMPRSS2 mainly in epithelial but also in mesenchymal and endothelial cells. Apart from the respiratory tract, no specific (histo-)morphologic alterations could be assigned to the SARS-CoV-2 infection.

scholarly journals Multisystemic cellular tropism of SARS-CoV-2 in autopsies of COVID-19 patients

2021 ◽  
Author(s):  
Dickson W.L. Wong ◽  
Barbara M. Klinkhammer ◽  
Sonja Djudjaj ◽  
Sophia Villwock ◽  
M. Cherelle Timm ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Virus Detection ◽  
Viral Rna ◽  
Transmission Electron ◽  
Formalin Fixed Paraffin ◽  
Organ Systems ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues

Background: Multiorgan tropism of SARS-CoV-2 has previously been shown for several major organs. Methods: We have comprehensively analyzed 25 different formalin-fixed paraffin-embedded (FFPE) tissues/organs from autopsies of fatal COVID-19 cases (n=8), using detailed histopathological assessment, detection of SARS-CoV-2 RNA using polymerase chain reaction and RNA in situ hybridization, viral protein using immunohistochemistry, and virus particles using transmission electron microscopy. Finally, we confirmed these findings in an independent external autopsy cohort (n=9). Findings: SARS-CoV-2 RNA was mainly localized in epithelial cells, endothelial and mesenchymal cells across all organs. Next to lung, trachea, kidney, heart, or liver, viral RNA was also found in tonsils, salivary glands, oropharynx, thyroid, adrenal gland, testicles, prostate, ovaries, small bowel, lymph nodes, skin and skeletal muscle. Viral RNA was predominantly found in cells expressing ACE2, TMPRSS2, or both. The SARS-CoV-2 replicating RNA was also detected in these organs. Immunohistochemistry and electron microscopy were not suitable for reliable and specific SARS-CoV-2 detection in autopsies. The findings were validated using in situ hybridization on external COVID-19 autopsy samples. Finally, apart from the lung, correlation of virus detection and histopathological assessment did not reveal any specific alterations that could be attributed to SARS-CoV-2. Conclusion: SARS-CoV-2 could be observed in virtually all organs, colocalizing with ACE2 and TMPRSS2 mainly in epithelial but also in mesenchymal and endothelial cells, and viral replication was found across all organ systems. Apart from the respiratory tract, no specific (histo-)morphologic alterations could be assigned to the SARS-CoV-2 infection.

scholarly journals Graphene oxide-quenching-based fluorescence in situ hybridization (G-FISH) to detect RNA in tissue: simple and fast tissue RNA diagnostics

2017 ◽  
Author(s):  
Do Won Hwang ◽  
Yoo Ri Choi ◽  
Dohyun Kim ◽  
Hye Yoon Park ◽  
Kyu Wan Kim ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Hippocampal Neurons ◽  
Rna Detection ◽  
Formalin Fixed Paraffin ◽  
Tumor Tissues ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues

AbstractFISH-based RNA detection in paraffin-embedded tissue can be challenging, with complicated procedures producing uncertain results and poor image quality. Here, we developed a robust RNA detection method based on graphene oxide (GO) quenching and recovery of fluorescence in situ hybridization (G-FISH) in formalin-fixed paraffin-embedded (FFPE) tissues. Using G-FISH technique, the long noncoding BC1 RNA, β-actin mRNA, miR-124a and miR-21 could be detected in the cytoplasm of a mouse brain, primary hippocampal neurons, and glioblastoma multiforme tumor tissues, respectively. G-FISH showed the increased BC1 RNA level in individual hippocampal neurons of Alzheimer’s disease brain. The fluorescence recovered by G-FISH correlated highly with the amount of miR-21, as measured by real time RT-PCR. We propose G-FISH as a simple, fast, inexpensive, and sensitive method for RNA detection, with very low background, which could be applied to a variety of researches or diagnostic purposes.

Rift Valley Fever Viral RNA Detection by In Situ Hybridization in Formalin-Fixed, Paraffin-Embedded Tissues

2019 ◽  
Vol 19 (7) ◽  
pp. 553-556 ◽  
Author(s):  
Izabela K. Ragan ◽  
Kaitlynn N. Schuck ◽  
Deepa Upreti ◽  
Lieza Odendaal ◽  
Juergen A. Richt ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Rift Valley ◽  
Rift Valley Fever ◽  
Viral Rna ◽  
Rna Detection ◽  
Valley Fever ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Canine Hepatitis Associated with Intrahepatic Bacteria in Three Dogs

2018 ◽  
Vol 54 (1) ◽  
pp. 65-70 ◽  
Author(s):  
Joon Im ◽  
Derek P. Burney ◽  
Sean P. McDonough ◽  
Brigid Nicholson ◽  
Adam Eatroff ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Case Report ◽  
Fluorescence In Situ Hybridization ◽  
Antimicrobial Therapy ◽  
Granulomatous Inflammation ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hematoxylin And Eosin ◽  
Formalin Fixed

ABSTRACT This case report describes the detection of intrahepatic bacteria in formalin-fixed paraffin-embedded histopathological sections from three dogs with neutrophilic, pyogranulomatous, or lymphoplasmacytic hepatitis and cholangiohepatitis. In each of these cases, eubacterial fluorescence in situ hybridization enabled colocalization of intrahepatic bacteria with neutrophilic and granulomatous inflammation in samples that were negative for bacteria when evaluated by routine hematoxylin and eosin histopathology augmented with histochemical stains. Positive responses to antimicrobial therapy were observed in of 2 out of 2 patients that were treated with antimicrobials. These findings suggest that eubacterial fluorescence in situ hybridization analysis of formalin-fixed paraffin-embedded histopathological sections is more sensitive than conventional histochemical stains for the diagnosis of bacteria-associated canine hepatitis.

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