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组织中严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的检测:逆转录定量聚合酶链反应(RT-qPCR)、原位RNA杂交和免疫组织化学的比较作用

Detection of SARS-CoV-2 in tissue: the comparative roles of RT-qPCR, in situ RNA hybridization, and immunohistochemistry.

作者信息

McHenry Austin, Iyer Krishna, Wang Jianhi, Liu Chen, Harigopal Malini

机构信息

Department of Pathology, School of Medicine, Yale University, New Haven, CT, United States.

出版信息

Expert Rev Mol Diagn. 2022 May;22(5):559-574. doi: 10.1080/14737159.2022.2085508. Epub 2022 Jun 10.

Abstract

INTRODUCTION

The emergence of SARS-CoV-2, the causative agent the COVID-19 pandemic, has led to a rapidly expanding arsenal of molecular diagnostic assays for the detection of viral material in tissue specimens.

AREAS COVERED

We review the value and shortcomings of available tissue-based assays for SARS-CoV-2 detection in formalin-fixed paraffin-embedded (FFPE) tissue, including immunohistochemistry, in situ hybridization, and quantitative reverse transcription PCR (RT-qPCR). The validation, accuracy, and comparative utility of each method is discussed. Subsequently, we identify commercially available antibodies which render the greatest specificity and reproducibility of staining in FFPE specimens.

EXPERT OPINION

We offer expert opinion on the efficacy of such techniques and guidance for future implementation, both clinical and experimental.

摘要

引言

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)作为导致新冠疫情的病原体出现后,用于检测组织标本中病毒物质的分子诊断检测方法迅速增加。

涵盖领域

我们综述了用于在福尔马林固定石蜡包埋(FFPE)组织中检测SARS-CoV-2的现有基于组织的检测方法的价值和缺点,包括免疫组织化学、原位杂交和定量逆转录聚合酶链反应(RT-qPCR)。讨论了每种方法的验证、准确性和比较实用性。随后,我们确定了在FFPE标本中染色具有最高特异性和可重复性的市售抗体。

专家意见

我们提供关于此类技术有效性的专家意见以及未来临床和实验实施的指导。

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