von Stillfried Saskia, Boor Peter
Institut für Pathologie, Universitätsklinik der RWTH Aachen, Pauwelsstr. 30, 52074, Aachen, Deutschland.
Medizinische Klinik II (Nephrologie und Immunologie), Universitätsklinik der RWTH Aachen, Pauwelsstr. 30, 52074, Aachen, Deutschland.
Pathologe. 2021 Mar;42(2):208-215. doi: 10.1007/s00292-021-00919-8. Epub 2021 Mar 1.
Analyses for the presence of SARS-CoV‑2 in the tissues of COVID-19 patients is important in order to improve our understanding of the disease pathophysiology for interpretation of diagnostic histopathological findings in autopsies, biopsies, or surgical specimens and to assess the potential for occupational infectious hazard.
In this review we identified 136 published studies in PubMed's curated literature database LitCovid on SARS-CoV‑2 detection methods in tissues and evaluated them regarding sources of error, specificity, and sensitivity of the methods, taking into account our own experience.
Currently, no sufficiently specific histomorphological alterations or diagnostic features for COVID-19 are known. Therefore, three approaches for SARS-CoV‑2 detection are used: RNA, proteins/antigens, or morphological detection by electron microscopy. In the preanalytical phase, the dominant source of error is tissue quality, especially the different intervals between sample collection and processing or fixation (and its duration) and specifically the interval between death and sample collection in autopsies. However, this information is found in less than half of the studies (e.g., in only 42% of autopsy studies). Our own experience and first studies prove the significantly higher sensitivity and specificity of RNA-based detection methods compared to antigen or protein detection by immunohistochemistry or immunofluorescence. Detection by electron microscopy is time consuming and difficult to interpret.
Different methods are available for the detection of SARS-CoV‑2 in tissue. Currently, RNA detection by RT-PCR is the method of choice. However, extensive validation studies and method harmonization are not available and are absolutely necessary.
分析2019冠状病毒病(COVID-19)患者组织中是否存在严重急性呼吸综合征冠状病毒2(SARS-CoV-2),对于增进我们对该疾病病理生理学的理解,以解释尸检、活检或手术标本中的诊断组织病理学发现,并评估职业感染风险具有重要意义。
在本综述中,我们在PubMed的精选文献数据库LitCovid中识别了136项关于组织中SARS-CoV-2检测方法的已发表研究,并结合我们自己的经验,对这些研究在误差来源、方法的特异性和敏感性方面进行了评估。
目前,尚无已知的针对COVID-19足够特异的组织形态学改变或诊断特征。因此,采用了三种检测SARS-CoV-2的方法:RNA、蛋白质/抗原或通过电子显微镜进行形态学检测。在分析前阶段,主要的误差来源是组织质量,尤其是样本采集与处理或固定之间的不同间隔(及其持续时间),特别是尸检中死亡与样本采集之间的间隔。然而,不到一半的研究中包含此信息(例如,仅42%的尸检研究中有)。我们自己的经验和初步研究证明,与免疫组织化学或免疫荧光检测抗原或蛋白质相比,基于RNA的检测方法具有显著更高的敏感性和特异性。电子显微镜检测耗时且难以解读。
有多种方法可用于检测组织中的SARS-CoV-2。目前,逆转录聚合酶链反应(RT-PCR)检测RNA是首选方法。然而,尚无广泛的验证研究且方法尚未统一,而这是绝对必要的。