中国 COVID-19 的实验室诊断:疑难病例回顾与分析。
Laboratory diagnosis of COVID-19 in China: A review of challenging cases and analysis.
机构信息
Department of Laboratory Medicine, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China; Graduate School, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China; Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases (BZ0447), Beijing, China.
School of Biomedical Sciences, Charles Sturt University, Leeds Parade, Orange, NSW, 2800, Australia; NSW Health Pathology, Orange Pathology Lab, Orange, NSW, 2800, Australia.
出版信息
J Microbiol Immunol Infect. 2021 Feb;54(1):17-26. doi: 10.1016/j.jmii.2020.10.004. Epub 2020 Oct 17.
Since the initial emergence of coronavirus disease 2019 (COVID-19) in Wuhan, Hubei province, China, a rapid spread of the disease occurred around the world, rising to become an international global health concern at pandemic level. In the face of this medical challenge threatening humans, the development of rapid and accurate methods for early screening and diagnosis of COVID-19 became crucial to containing the emerging public health threat, and prevent further spread within the population. Despite the large number of COVID-19 confirmed cases in China, some problematic cases with inconsistent laboratory testing results, were reported. Specifically, a high false-negative rate of 41% on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection by real-time reverse transcription-polymerase chain reaction (qRT-PCR) assays was observed in China. Although serological testing has been applied worldwide as a complementary method to help identify SARS-CoV-2, several limitations on its use have been reported in China. Therefore, the use of both qRT-PCR and serological testing in the diagnosis of COVID-19 in China and elsewhere, presented considerable challenges, but when used in combination, can be valuable tools in the fight against COVID-19. In this review, we give an overview of the advantages and disadvantages of different molecular techniques for SARS-CoV-2 detection that are currently used in several labs, including qRT-PCR, gene sequencing, loop-mediated isothermal amplification (LAMP), nucleic acid mass spectrometry (MS), and gene editing technique based on clustered regularly interspaced short palindromic repeats (CRISPR/Cas13) system. Then we mainly review and analyze some causes of false-negative qRT-PCR results, and how to resolve some of the diagnostic dilemma.
自 2019 年冠状病毒病(COVID-19)在中国湖北省武汉市首次出现以来,该疾病在全球迅速传播,上升为大流行级别的国际公共卫生关注。面对这一威胁人类的医学挑战,开发快速、准确的 COVID-19 早期筛查和诊断方法对于遏制这一新兴的公共卫生威胁并防止其在人群中进一步传播至关重要。尽管中国 COVID-19 确诊病例数量众多,但仍有一些实验室检测结果不一致的疑难病例报告。具体来说,中国实时逆转录聚合酶链反应(qRT-PCR)检测严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)的假阴性率高达 41%。虽然血清学检测已在全球范围内作为一种补充方法用于帮助识别 SARS-CoV-2,但在中国也报告了其使用的一些局限性。因此,qRT-PCR 和血清学检测在中国和其他地方用于 COVID-19 的诊断都面临着相当大的挑战,但联合使用时,可成为抗击 COVID-19 的有价值工具。在这篇综述中,我们概述了目前在多个实验室中使用的 SARS-CoV-2 检测的不同分子技术的优缺点,包括 qRT-PCR、基因测序、环介导等温扩增(LAMP)、核酸质谱(MS)和基于簇状规则间隔短回文重复序列(CRISPR/Cas13)系统的基因编辑技术。然后,我们主要回顾和分析了一些导致 qRT-PCR 结果假阴性的原因,并探讨了如何解决一些诊断难题。