Caniego-Casas Tamara, Martínez-García Laura, Alonso-Riaño Marina, Pizarro David, Carretero-Barrio Irene, Martínez-de-Castro Nilda, Ruz-Caracuel Ignacio, de Pablo Raúl, Saiz Ana, Royo Rosa Nieto, Santiago Ana, Rosas Marta, Rodríguez-Peralto José L, Pérez-Mies Belén, Galán Juan C, Palacios José
Pathology Department, Hospital Universitario Ramón y Cajal, Madrid, Spain.
Instituto Ramón y Cajal for Health Research (IRYCIS), Madrid, Spain.
Front Microbiol. 2022 Jan 31;13:824967. doi: 10.3389/fmicb.2022.824967. eCollection 2022.
The exact role of viral replication in patients with severe COVID-19 has not been extensively studied, and it has only been possible to demonstrate the presence of replicative virus for more than 3 months in a few cases using different techniques. Our objective was to study the presence of RNA SARS-CoV-2 in autopsy samples of patients who died from COVID-19 long after the onset of symptoms. Secondary superimposed pulmonary infections present in these patients were also studied. We present an autopsy series of 27 COVID-19 patients with long disease duration, where pulmonary and extrapulmonary samples were obtained. In addition to histopathological analysis, viral genomic RNA (gRNA) and viral subgenomic RNA (sgRNA) were detected using RT-PCR and hybridization, and viral protein was detected using immunohistochemistry. This series includes 26 adults with a median duration of 39 days from onset of symptoms to death (ranging 9-108 days), 92% of them subjected to immunomodulatory therapy, and an infant patient. We detected gRNA in the lung of all but one patient, including those with longer disease duration. SgRNA was detected in 11 out of 17 patients (64.7%) with illness duration up to 6 weeks and in 3 out of 9 patients (33.3%) with more than 6 weeks of disease progression. Viral protein was detected using immunohistochemistry and viral mRNA was detected using hybridization in 3 out of 4 adult patients with illness duration of <2 weeks, but in none of the 23 adult patients with an illness duration of >2 weeks. A remarkable result was the detection of viral protein, gRNA and sgRNA in the lung cells of the pediatric patient after 95 days of illness. Additional pulmonary infections included: 9 acute bronchopneumonia, 2 aspergillosis, 2 cytomegalovirus, and 1 BK virus infection. These results suggest that in severe COVID-19, SARS-CoV-2 could persist for longer periods than expected, especially in immunocompromised populations, contributing to the persistence of chronic lung lesions. Additional infections contribute to the fatal course of the disease.
病毒复制在重症新冠肺炎患者中的确切作用尚未得到广泛研究，而且仅在少数病例中通过不同技术才得以证实在症状出现三个多月后仍存在复制型病毒。我们的目的是研究在症状出现很久后死于新冠肺炎的患者尸检样本中是否存在新冠病毒RNA。对这些患者中出现的继发性叠加肺部感染也进行了研究。我们展示了一个针对27例病程较长的新冠肺炎患者的尸检系列，获取了其肺部和肺外样本。除了组织病理学分析外，还使用逆转录聚合酶链反应（RT-PCR）和杂交检测病毒基因组RNA（gRNA）和病毒亚基因组RNA（sgRNA），并使用免疫组织化学检测病毒蛋白。该系列包括26名成年人，从症状出现到死亡的中位时间为39天（范围9 - 108天），其中92%接受了免疫调节治疗，还有一名婴儿患者。除一名患者外，我们在所有患者的肺部检测到了gRNA，包括病程较长的患者。在病程长达6周的17名患者中有11名（64.7%）检测到sgRNA，在病程超过6周的9名患者中有3名（33.3%）检测到sgRNA。在病程<2周的4名成年患者中有3名通过免疫组织化学检测到病毒蛋白，通过杂交检测到病毒mRNA，但在病程>2周的23名成年患者中均未检测到。一个显著的结果是在患病95天后在这名儿科患者的肺细胞中检测到了病毒蛋白、gRNA和sgRNA。额外的肺部感染包括：9例急性支气管肺炎、2例曲霉菌病、2例巨细胞病毒感染和1例BK病毒感染。这些结果表明，在重症新冠肺炎中，新冠病毒可能比预期持续更长时间，尤其是在免疫功能低下人群中，这导致了慢性肺部病变的持续存在。额外的感染促成了疾病的致命进程。
