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两分钟液基样本制备法用于快速 SARS-CoV-2 实时 PCR 筛查:一项多中心评估。

A two minute liquid based sample preparation for rapid SARS-CoV2 real-time PCR screening: A multicentre evaluation.

机构信息

Leiden University Medical Center, Department of Medical Microbiology, Leiden, the Netherlands.

Maasstad Hospital, Medical Microbiology Laboratory, Rotterdam, the Netherlands.

出版信息

J Clin Virol. 2021 Feb;135:104720. doi: 10.1016/j.jcv.2020.104720. Epub 2020 Dec 31.

Abstract

BACKGROUND

Apart from major health concerns associated to the SARS-coronavirus-2 (SARS-CoV-2) pandemic, also the diagnostic workflow encountered serious problems. Limited availability of kit components, buffers and even plastics has resulted in suboptimal testing procedures worldwide. Alternative workflows have been implemented to overcome these difficulties. Recently a liquid based sample prep has been launched as solution to overcome limitations in relation to nucleic acid extraction.

OBJECTIVE

Multicenter evaluation of the QIAprep&amp Viral RNA UM kit (QIA P&A) for rapid sample preparation and real-time PCR detection of SARS-CoV-2 in comparison to standardized laboratory testing methods.

STUDY DESIGN

Selected samples of the routine diagnostic workflow at Clinical Microbiology Laboratories of four Dutch hospitals have been subjected to the rapid QIA P&A protocol and the results have been compared to routine diagnostic data.

RESULTS

Combining results of manual and automated procedures, a total of 377 clinical samples of which 202 had been tested positive with a wide range of C values, showed almost complete concordance in the QIA P&A assay for samples up to C values of 33 with one exception of C 31. Prospectively 60 samples were tested and also showed 100 % concordance with 5 positives. The method has been automated by two centres.

CONCLUSIONS

Despite an input of only 8 μL of clinical sample, the QIA P&A kit showed good performance for sample preparation and amplification of SARS-CoV-2 and can contribute as a rapid molecular testing strategy in managing the CoV-2 pandemic.

摘要

背景

除了与 SARS-CoV-2(SARS-CoV-2)大流行相关的主要健康问题外,诊断工作流程也遇到了严重的问题。试剂盒组件、缓冲液甚至塑料的有限供应导致全球检测程序都不够理想。已经实施了替代工作流程来克服这些困难。最近推出了一种基于液体的样本制备方法,作为克服与核酸提取相关的限制的解决方案。

目的

对 QIAprep&Viral RNA UM 试剂盒(QIA P&A)进行多中心评估,用于快速样品制备和实时 PCR 检测 SARS-CoV-2,与标准化实验室检测方法进行比较。

研究设计

对四家荷兰医院临床微生物学实验室常规诊断工作流程中的选定样本进行了快速 QIA P&A 方案处理,并将结果与常规诊断数据进行了比较。

结果

结合手动和自动程序的结果,总共 377 个临床样本中,有 202 个样本的 C 值范围很广,检测结果呈阳性,在 QIA P&A 检测中,除了 C 值为 31 的样本外,对于 C 值高达 33 的样本,几乎完全一致。前瞻性地对 60 个样本进行了测试,其中 5 个阳性样本的一致性为 100%。该方法已由两个中心实现自动化。

结论

尽管仅输入 8 μL 的临床样本,QIA P&A 试剂盒在 SARS-CoV-2 的样品制备和扩增方面表现出良好的性能,并且可以作为管理 CoV-2 大流行的快速分子检测策略。

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