RT-qPCR investigation of post-mortem tissues during COVID-19
Berdygulova Z. Maltseva E. Perfilyeva Y. Nizkorodova A. Zhigailov A. Naizabayeva D. Ostapchuk Y.O. Kuatbekova S. Dosmagambet Z. Kuatbek M. Bissenbay A. Cherusheva A. Mashzhan A. Abdolla N. Ashimbekov S. Ismagulova G. Dmitrovskiy A. Mamadaliyev S. Skiba Y.
2024University of South Bohemia in Ceske Budejovice Faculty of Health and Social Sciences
Journal of Applied Biomedicine
2024#22Issue 2115 - 122 pp.
In 2020, there were numerous cases in Kazakhstan with clinical symptoms of COVID-19 but negative PCR results in nasopharyngeal and oropharyngeal swabs. The diagnosis was confirmed clinically and by CT scans (computed tomography). The problem with such negative PCR results for SARS-CoV-2 infection confirmation still exists and indicates the need to confirm the diagnosis in the bronchoalveolar lavage in such cases. There is also a lack of information about confirmation of SARS-CoV-2 infection in deceased patients. In this study, various tissue materials, including lungs, bronchi, and trachea, were examined from eight patients who died, presumably from SARS-CoV-2 infection, between 2020 and 2022. Naso/oropharyngeal swabs taken from these patients in hospitals tested PCR negative for SARS-CoV-2. This study presents a modified RNA isolation method based on a comparison of the most used methods for RNA isolation in laboratories: QIAamp Viral RNA Mini Kit and TRIzol-based method. This modified nucleic acid extraction protocol can be used to confirm SARS-CoV-2 infection by RT-qPCR in the tissues of deceased patients in disputed cases. RT-qPCR with RNA of SARS-CoV-2 re-extracted with such method from post-mortem tissues that were stored at –80 °C for more than 32 months still demonstrated high-yielding positive results.
COVID-19 , Post-mortem tissues , RNA extraction , RT-qPCR , SARS-CoV-2
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Almaty Branch of the National Center for Biotechnology, Central Reference Laboratory, Almaty, Kazakhstan
M. A. Aitkhozhin Institute of Molecular Biology and Biochemistry, Almaty, Kazakhstan
Tethys Scientific Society, Almaty, Kazakhstan
Asfendiyarov Kazakh National Medical University, Almaty, Kazakhstan
Almaty Branch of the National Center for Biotechnology
M. A. Aitkhozhin Institute of Molecular Biology and Biochemistry
Tethys Scientific Society
Asfendiyarov Kazakh National Medical University
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