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RT-qPCR investigation of post-mortem tissues during COVID-19

Z. Berdygulova, E. Maltseva, Y. Perfilyeva, A. Nizkorodova, A. Zhigailov, D. Naizabayeva, Y. O. Ostapchuk, S. Kuatbekova, Z. Dosmagambet, M. Kuatbek, A. Bissenbay, A. Cherusheva, A. Mashzhan, N. Abdolla, S. Ashimbekov, G. Ismagulova, A....

. 2024 ; 22 (2) : 115-122. [pub] 20240621

Language English Country Czech Republic

Document type Journal Article

Persistent link   https://www.medvik.cz/link/bmc24011942

Grant support
AP09259103 Committee of Science of the Ministry of Science and Higher Education of the Republic of Kazakhstan - Kazakhstan
COVID-19 Committee of Science of the Ministry of Science and Higher Education of the Republic of Kazakhstan - Kazakhstan

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.

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