Demyashkin G A, Gorokhov K R, Nikitin P V, Zorin I A, Voznesenskaya S I
National Medical Research Centre of Radiology, Obninsk, Russia.
I.M. Sechenov First Moscow State Medical University, Moscow, Russia.
Arkh Patol. 2023;85(3):23-28. doi: 10.17116/patol20238503123.
Background. The novel coronavirus infection (COVID-19) often manifests in children as diarrhea, vomiting, abdominal pain, and some children develop acute appendicitis. To elucidate the role of SARS-CoV-2 in the development of acute appendicitis, a more detailed study of the presence of its genetic material in the tissue of the appendix.
Determination of SARS-CoV-2 RNA in appendices of children with COVID-19 by fluorescence in situ hybridization (FISH).
A retrospective analysis of case histories and morphological analysis using FISH of appendices of pediatric patients with established clinical diagnosis of acute appendicitis and confirmed infection with SARS-CoV-2 was performed. The material was divided into 3 groups: 1st -appendices obtained during appendectomy in children with established clinical diagnosis of «coronavirus infection» (COVID-19, PCR+) (=42; mean age 10.8 years); 2nd - appendices of children (=55; mean age 9.7 years) with acute appendicitis obtained before the onset of the COVID-19 pandemic; 3rd (control) group (=38; mean age 10.3 years) - autopsy material of the appendices (intact).
In all samples of the appendices of the 1st group, a positive SARS-CoV-2 viral RNA signal was noted in the cytoplasm of most epithelial cells and single immunocompetent cells. The signal intensity remained the same in all slides, regardless of age. In all samples obtained from patients without COVID-19 (groups 2 and 3), confocal microscopy did not reveal a signal, which indicates successful adaptation of the FISH method in this study and excludes the false positive results.
In the epithelium of the appendices of children of different age with COVID-19, the FISH method revealed SARS-CoV-2 RNA, which does not exclude the association between viral invasion and the development of acute appendicitis.
背景。新型冠状病毒感染（COVID-19）在儿童中常表现为腹泻、呕吐、腹痛，部分儿童会发展为急性阑尾炎。为阐明严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在急性阑尾炎发病中的作用，需对其遗传物质在阑尾组织中的存在情况进行更详细研究。
通过荧光原位杂交（FISH）检测COVID-19患儿阑尾中SARS-CoV-2 RNA。
对已确诊临床诊断为急性阑尾炎且确诊感染SARS-CoV-2的儿科患者的阑尾进行病例史回顾分析及FISH形态学分析。材料分为3组：第1组 - 在临床诊断为“冠状病毒感染”（COVID-19，PCR阳性）的儿童阑尾切除术中获取的阑尾（=42例；平均年龄10.8岁）；第2组 - 在COVID-19大流行开始前获得的患有急性阑尾炎的儿童阑尾（=55例；平均年龄9.7岁）；第3组（对照组）（=38例；平均年龄10.3岁） - 完整阑尾的尸检材料。
在第1组阑尾的所有样本中，大多数上皮细胞和单个免疫活性细胞的细胞质中均检测到阳性SARS-CoV-2病毒RNA信号。所有玻片上信号强度均相同，与年龄无关。在所有未感染COVID-19患者（第2组和第3组）的样本中，共聚焦显微镜未发现信号，这表明本研究中FISH方法适用且排除了假阳性结果。
FISH方法在不同年龄COVID-19患儿阑尾上皮中检测到SARS-CoV-2 RNA，这并不排除病毒侵袭与急性阑尾炎发病之间的关联。
