{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,20]],"date-time":"2026-02-20T16:37:03Z","timestamp":1771605423891,"version":"3.50.1"},"reference-count":47,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2023,5,7]],"date-time":"2023-05-07T00:00:00Z","timestamp":1683417600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Higher Education of the Russian Federation","award":["075-15-2022-301"],"award-info":[{"award-number":["075-15-2022-301"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background: Few studies have compared COVID-19 patients from different waves. This study aims to conduct a clinical and morphological analysis of patients who died from COVID-19 during four waves. Methods: The study involved 276 patients who died from COVID-19 during four waves, including 77 patients in the first wave, 119 patients in the second wave, and 78 patients in the third wave. We performed a histological examination of myocardium samples from autopsies and additionally analyzed the samples by PCR. We conducted immunohistochemistry of the myocardium for 21 samples using antibodies against CD3, CD45, CD8, CD68, CD34, Ang1, VWF, VEGF, HLA-DR, MHC1, C1q, enteroviral VP1, and SARS-CoV-2 spike protein. We also did immunofluorescent staining of three myocardial specimens using VP1\/SARS-CoV-2 antibody cocktails. Further, we ran RT-ddPCR analysis for 14 RNA samples extracted from paraffin-embedded myocardium. Electron microscopic studies of the myocardium were also performed for two samples from the fourth wave. Results: Among the 276 cases, active myocarditis was diagnosed in 5% (15\/276). Of these cases, 86% of samples expressed VP1, and individual cells contained SARS-CoV-2 spike protein in 22%. Immunofluorescence confirmed the co-localization of VP1 and SARS-CoV-2 spike proteins. ddPCR did not confidently detect SARS-CoV-2 RNA in the myocardium in any myocarditis cases. However, the myocardium sample from wave IV detected a sub-threshold signal of SARS-CoV-2 by qPCR, but myocarditis in this patient was not confirmed. Electron microscopy showed several single particles similar to SARS-CoV-2 virions on the surface of the endothelium of myocardial vessels. A comparison of the cardiovascular complication incidence between three waves revealed that the incidence of hemorrhage (48 vs. 24 vs. 17%), myocardial necrosis (18 vs. 11 vs. 4%), blood clots in the intramural arteries (12 vs. 7 vs. 0%), and myocarditis (19 vs. 1 vs. 6%) decreased over time, and CD8-T-killers appeared. Immunohistochemistry confirmed the presence of endotheliitis in all 21 studied cases. Conclusions: This study compared myocardial damage in patients who died during three COVID-19 waves and showed a decrease in the incidence of endotheliitis complications (thrombosis, hemorrhage, necrosis) and myocarditis over time. However, the connection between myocarditis and SARS-CoV-2 infection remains unproven.<\/jats:p>","DOI":"10.3390\/diagnostics13091645","type":"journal-article","created":{"date-parts":[[2023,5,8]],"date-time":"2023-05-08T01:32:53Z","timestamp":1683509573000},"page":"1645","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Comparative Study of the Myocardium of Patients from Four COVID-19 Waves"],"prefix":"10.3390","volume":"13","author":[{"given":"Lubov Borisovna","family":"Mitrofanova","sequence":"first","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6175-8403","authenticated-orcid":false,"given":"Igor Aleksandrovich","family":"Makarov","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrey Nikolaevich","family":"Gorshkov","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"},{"name":"Smorodintsev Research Institute of Influenza, St. Petersburg 197376, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrey Leonidovich","family":"Runov","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"},{"name":"D.I. Mendeleyev Institute for Metrology, St. Petersburg 190005, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maxim Sergeevich","family":"Vonsky","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"},{"name":"D.I. Mendeleyev Institute for Metrology, St. Petersburg 190005, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Mikhailovna","family":"Pisareva","sequence":"additional","affiliation":[{"name":"Smorodintsev Research Institute of Influenza, St. Petersburg 197376, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrey Borisovich","family":"Komissarov","sequence":"additional","affiliation":[{"name":"Smorodintsev Research Institute of Influenza, St. Petersburg 197376, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Taiana Alekseevna","family":"Makarova","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qingli","family":"Li","sequence":"additional","affiliation":[{"name":"East China Normal University, Shanghai 200241, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tatiana Leonidovna","family":"Karonova","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8169-7812","authenticated-orcid":false,"given":"Alexandra Olegovna","family":"Konradi","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Evgeny Vladimirovich","family":"Shlaykhto","sequence":"additional","affiliation":[{"name":"Almazov National Medical Research Centre, St. Petersburg 197341, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,5,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.jinf.2020.10.033","article-title":"First and second COVID-19 waves in Japan: A comparison of disease severity and characteristics","volume":"82","author":"Saito","year":"2021","journal-title":"J. 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