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龈沟液中 SARS-CoV-2 的检测。

SARS-CoV-2 Detection in Gingival Crevicular Fluid.

机构信息

Unit of Periodontics, Oral Health Sciences Centre, Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India.

Department of Internal Medicine, PGIMER, Chandigarh, India.

出版信息

J Dent Res. 2021 Feb;100(2):187-193. doi: 10.1177/0022034520970536. Epub 2020 Nov 2.

Abstract

Understanding the pathophysiology of the coronavirus disease 2019 (COVID-19) infection remains a significant challenge of our times. The gingival crevicular fluid being representative of systemic status and having a proven track record of detecting viruses and biomarkers forms a logical basis for evaluating the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The study aimed to assess gingival crevicular fluid (GCF) for evidence of SARS-CoV-2 in 33 patients who were deemed to be COVID-19 positive upon nasopharyngeal sampling. An attempt was also made to comparatively evaluate it with saliva in terms of its sensitivity, as a diagnostic fluid for SARS-CoV-2. GCF and saliva samples were collected from 33 COVID-19-confirmed patients. Total RNA was extracted using NucliSENS easyMAG (bioMérieux) and eluted in the elution buffer. Envelope gene ( gene) of SARS-CoV-2 and human RNase P gene as internal control were detected in GCF samples by using the TRUPCR SARS-CoV-2 RT qPCR kit V-2.0 (I) in an Applied Biosystems 7500 real-time machine. A significant majority of both asymptomatic and mildly symptomatic patients exhibited the presence of the novel coronavirus in their GCF samples. Considering the presence of SARS-CoV-2 RNA in the nasopharyngeal swab sampling as gold standard, the sensitivity of GCF and saliva, respectively, was 63.64% (confidence interval [CI], 45.1% to 79.60%) and 64.52% (CI, 45.37% to 80.77%). GCF was found to be comparable to saliva in terms of its sensitivity to detect SARS-CoV-2. Saliva samples tested positive in 3 of the 12 patients whose GCF tested negative, and likewise GCF tested positive for 2 of the 11 patients whose saliva tested negative on real-time reverse transcription polymerase chain reaction. The results establish GCF as a possible mode of transmission of SARS-CoV-2, which is the first such report in the literature, and also provide the first quantifiable evidence pointing toward a link between the COVID-19 infection and oral health.

摘要

了解 2019 年冠状病毒病(COVID-19)感染的病理生理学仍然是我们这个时代的重大挑战。牙龈沟液是系统性状态的代表,并且已经有检测病毒和生物标志物的可靠记录,这为评估严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)的存在提供了合理的依据。本研究旨在评估 33 名经鼻咽取样被认为 COVID-19 阳性的患者的牙龈沟液(GCF)中是否存在 SARS-CoV-2。还尝试将其与唾液进行比较,以评估其作为 SARS-CoV-2 诊断液的敏感性。从 33 名 COVID-19 确诊患者中采集 GCF 和唾液样本。使用 NucliSENS easyMAG(bioMérieux)提取总 RNA,并在洗脱缓冲液中洗脱。使用 TRUPCR SARS-CoV-2 RT qPCR 试剂盒 V-2.0(I)(Applied Biosystems 7500 实时机器)在 GCF 样本中检测 SARS-CoV-2 的包膜基因(基因)和人 RNA 聚合酶 P 基因作为内部对照。相当大比例的无症状和轻度症状患者的 GCF 样本中均存在新型冠状病毒。考虑到鼻咽拭子采样中 SARS-CoV-2 RNA 的存在作为金标准,GCF 和唾液的灵敏度分别为 63.64%(置信区间[CI],45.1%至 79.60%)和 64.52%(CI,45.37%至 80.77%)。就检测 SARS-CoV-2 的灵敏度而言,GCF 与唾液相当。在 12 名 GCF 检测阴性的患者中,有 3 名唾液样本检测阳性,同样在 11 名 GCF 检测阴性的患者中,有 2 名唾液样本检测阳性。实时逆转录聚合酶链反应。这些结果确立了 GCF 是 SARS-CoV-2 的一种可能传播方式,这是文献中的首次报道,也提供了 COVID-19 感染与口腔健康之间存在关联的首个可量化证据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/25fb/7817989/db1f1a6cccc5/10.1177_0022034520970536-fig1.jpg

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