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Digital PCR to quantify ChAdOx1 nCoV-19 copies in blood and tissues

Anita Badbaran, Reiner Mailer, Christine Dahlke, Jannis Woens, Anahita Fathi, Sibylle C. Mellinghoff, Thomas Renné, Marylyn M. Addo, View ORCID ProfileKristoffer Riecken, View ORCID ProfileBoris Fehse
doi: https://doi.org/10.1101/2021.05.28.446155
Anita Badbaran
1Dept. of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
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Reiner Mailer
2Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
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Christine Dahlke
3Division of Infectious Diseases, 1st Department of Medicine, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
4Department for Clinical Immunology of Infectious Diseases, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany
5German Center for Infection Research (DZIF), Partner Site Hamburg-Lübeck-Borstel-Riems, Germany
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Jannis Woens
1Dept. of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
6Research Dept. Cell and Gene Therapy, UKE, Hamburg, Germany
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Anahita Fathi
3Division of Infectious Diseases, 1st Department of Medicine, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
4Department for Clinical Immunology of Infectious Diseases, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany
5German Center for Infection Research (DZIF), Partner Site Hamburg-Lübeck-Borstel-Riems, Germany
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Sibylle C. Mellinghoff
3Division of Infectious Diseases, 1st Department of Medicine, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
4Department for Clinical Immunology of Infectious Diseases, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany
5German Center for Infection Research (DZIF), Partner Site Hamburg-Lübeck-Borstel-Riems, Germany
7University of Cologne, Faculty of Medicine and University Hospital Cologne, Dept. I of Internal Medicine, Excellence Center for Medical Mycology (ECMM), Cologne, Germany
8German Centre for Infection Research (DZIF), Partner Site Bonn-Cologne, Cologne, Germany
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Thomas Renné
2Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
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Marylyn M. Addo
3Division of Infectious Diseases, 1st Department of Medicine, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
4Department for Clinical Immunology of Infectious Diseases, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany
5German Center for Infection Research (DZIF), Partner Site Hamburg-Lübeck-Borstel-Riems, Germany
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Kristoffer Riecken
1Dept. of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
6Research Dept. Cell and Gene Therapy, UKE, Hamburg, Germany
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  • ORCID record for Kristoffer Riecken
Boris Fehse
1Dept. of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf (UKE), Hamburg
5German Center for Infection Research (DZIF), Partner Site Hamburg-Lübeck-Borstel-Riems, Germany
6Research Dept. Cell and Gene Therapy, UKE, Hamburg, Germany
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  • ORCID record for Boris Fehse
  • For correspondence: fehse@uke.de
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Abstract

Vaccination with the adenoviral-vector based Astra Zeneca ChAdOx1 nCov-19 vaccine is efficient and safe. However, in rare cases vaccinated individuals developed life-threatening thrombotic complications, including thrombosis in cerebral sinus and splanchnic veins. Monitoring of the applied vector in vivo represents an important precondition to study the molecular mechanisms underlying vaccine-driven adverse effects now referred to as vaccine-induced immune thrombotic thrombocytopenia (VITT). We previously have shown that digital PCR is an excellent tool to quantify transgene copies in vivo. Here we present a highly sensitive digital PCR for in-situ quantification of ChAdOx1 nCoV-19 copies. Using this method, we quantified vector copies in human serum 24, 72 and 168 hours post vaccination, and in a variety of murine tissues in an experimental vaccination model 30 minutes post injection. We describe a method for high-sensitivity quantitative detection of ChAdOx1 nCoV-19 with possible implications to elucidate the mechanisms of severe ChAdOx1 nCov-19 vaccine complications.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 28, 2021.
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Digital PCR to quantify ChAdOx1 nCoV-19 copies in blood and tissues
Anita Badbaran, Reiner Mailer, Christine Dahlke, Jannis Woens, Anahita Fathi, Sibylle C. Mellinghoff, Thomas Renné, Marylyn M. Addo, Kristoffer Riecken, Boris Fehse
bioRxiv 2021.05.28.446155; doi: https://doi.org/10.1101/2021.05.28.446155
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Digital PCR to quantify ChAdOx1 nCoV-19 copies in blood and tissues
Anita Badbaran, Reiner Mailer, Christine Dahlke, Jannis Woens, Anahita Fathi, Sibylle C. Mellinghoff, Thomas Renné, Marylyn M. Addo, Kristoffer Riecken, Boris Fehse
bioRxiv 2021.05.28.446155; doi: https://doi.org/10.1101/2021.05.28.446155

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