Ohio State University Comprehensive Cancer Center, Columbus, OH, United States of America; Gnome Diagnostics, Powell, OH, United States of America.
Weill Cornell Medicine, New York, NY, United States of America.
Ann Diagn Pathol. 2020 Oct;48:151565. doi: 10.1016/j.anndiagpath.2020.151565. Epub 2020 Jul 7.
Infection by SARS-CoV-2 commonly begins in the nasopharynx, and the cytologic and molecular correlates are not characterized. Fifty-eight cytologic preps (20 oral and 38 from the nasopharynx) were obtained from ten patients and analyzed in a blinded fashion for SARS-CoV-2 spike and envelope protein by immunohistochemistry and viral RNA by in situ hybridization. qRTPCR identified three positive cases and seven controls; the three cases reported mild symptoms that resolved in 2-3 days. Blinded analyses confirmed the presence of the SARS-CoV-2 spike and envelope proteins and viral RNA in the three cases and viral absence in the seven controls. A signal for the positive cases was evident in each nasopharyngeal and none of the oral samples. Viral RNA/proteins localized exclusively to glandular cells and was present in high copy number. Blinded analysis of the cytology documented that the glandular cells infected by SARS-CoV-2 showed marked degeneration with ciliocytophthoria; viral inclusions were not evident. Co-expression analysis showed viral infected cells had increased apoptosis, marked by strong expression of activated caspase 3. Weekly serial testing of two of the cases showed persistence of productive viral infection for up to 2 weeks after symptom onset. It is concluded that the target cell of SARS-CoV-2 in the head and neck region is the glandular cell of the nasal passages, that viral infection is lytic and associated with high copy number that facilitates viral spread. The method outlines a simple, rapid test for productive SARS-CoV-2 based on immunohistochemistry or in situ hybridization of the glandular cells from the nasopharynx.
SARS-CoV-2 感染通常始于鼻咽部，其细胞学和分子相关性尚不清楚。我们对 10 名患者的 58 份细胞学标本（20 份口腔标本和 38 份鼻咽部标本）进行了盲法分析，通过免疫组织化学法检测 SARS-CoV-2 的刺突蛋白和包膜蛋白，通过原位杂交法检测病毒 RNA。qRTPCR 鉴定了 3 个阳性病例和 7 个对照病例；这 3 个病例报告的症状轻微，在 2-3 天内缓解。盲法分析证实了这 3 个病例存在 SARS-CoV-2 的刺突蛋白和包膜蛋白以及病毒 RNA，而 7 个对照病例则不存在病毒。在每个鼻咽部标本中都能明显检测到阳性病例的信号，而在口腔标本中则没有。病毒 RNA/蛋白仅定位于腺细胞，且拷贝数很高。细胞学的盲法分析记录了受 SARS-CoV-2 感染的腺细胞出现明显的退行性变化，伴有纤毛细胞空泡化；没有观察到病毒包涵体。病毒感染细胞的共表达分析显示细胞凋亡增加，这一特征表现为激活的 caspase 3 强表达。对其中 2 例患者的每周连续检测显示，在症状出现后长达 2 周的时间里，持续存在有复制能力的病毒感染。结论是，SARS-CoV-2 在头颈部的靶细胞是鼻腔的腺细胞，病毒感染是裂解性的，且病毒载量高，有利于病毒传播。该方法概述了一种基于鼻咽部腺细胞的免疫组织化学或原位杂交的简单、快速的 SARS-CoV-2 有复制能力检测方法。
