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SARS-CoV、MERS-CoV 和 SARS-CoV-2(2019-nCoV)的实验室检测:现状、挑战和对策。

Laboratory testing of SARS-CoV, MERS-CoV, and SARS-CoV-2 (2019-nCoV): Current status, challenges, and countermeasures.

机构信息

National Center for Clinical Laboratories, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, China.

Beijing Engineering Research Center of Laboratory Medicine, Beijing Hospital, Beijing, China.

出版信息

Rev Med Virol. 2020 May;30(3):e2106. doi: 10.1002/rmv.2106. Epub 2020 Apr 17.

Abstract

Emerging and reemerging infectious diseases are global public concerns. With the outbreak of unknown pneumonia in Wuhan, China in December 2019, a new coronavirus, SARS-CoV-2 has been attracting tremendous attention. Rapid and accurate laboratory testing of SARS-CoV-2 is essential for early discovery, early reporting, early quarantine, early treatment, and cutting off epidemic transmission. The genome structure, transmission, and pathogenesis of SARS-CoV-2 are basically similar to SARS-CoV and MERS-CoV, the other two beta-CoVs of medical importance. During the SARS-CoV and MERS-CoV epidemics, a variety of molecular and serological diagnostic assays were established and should be referred to for SARS-CoV-2. In this review, by summarizing the articles and guidelines about specimen collection, nucleic acid tests (NAT) and serological tests for SARS-CoV, MERS-CoV, and SARS-CoV-2, several suggestions are put forward to improve the laboratory testing of SARS-CoV-2. In summary, for NAT: collecting stool and blood samples at later periods of illness to improve the positive rate if lower respiratory tract specimens are unavailable; increasing template volume to raise the sensitivity of detection; putting samples in reagents containing guanidine salt to inactivate virus as well as protect RNA; setting proper positive, negative and inhibition controls to ensure high-quality results; simultaneously amplifying human RNase P gene to avoid false-negative results. For antibody test, diverse assays targeting different antigens, and collecting paired samples are needed.

摘要

新发和再发传染病是全球公共卫生关注的焦点。2019 年 12 月,中国武汉爆发不明原因肺炎,一种新型冠状病毒 SARS-CoV-2 引起了极大关注。快速准确地检测 SARS-CoV-2 对于早期发现、早期报告、早期隔离、早期治疗和阻断疫情传播至关重要。SARS-CoV-2 的基因组结构、传播和发病机制与 SARS-CoV 和 MERS-CoV 基本相似,这两种病毒也是具有医学重要性的另外两种β冠状病毒。在 SARS-CoV 和 MERS-CoV 流行期间,建立了多种分子和血清学诊断检测方法,可参考用于 SARS-CoV-2 的检测。在本综述中,通过总结有关 SARS-CoV、MERS-CoV 和 SARS-CoV-2 的标本采集、核酸检测(NAT)和血清学检测的文章和指南,提出了一些改进 SARS-CoV-2 实验室检测的建议。总的来说,对于 NAT:如果无法获得下呼吸道标本,在疾病后期采集粪便和血液样本以提高阳性率;增加模板量以提高检测灵敏度;将样本置于含胍盐的试剂中以灭活病毒并保护 RNA;设置适当的阳性、阴性和抑制对照,以确保高质量的结果;同时扩增人 RNase P 基因,以避免假阴性结果。对于抗体检测,需要针对不同抗原的多种检测方法,并采集配对样本。

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