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逆转录重组酶辅助扩增检测试剂盒结合侧向流试纸条用于快速检测 2019 新型冠状病毒。

Reverse Transcription Recombinase-Aided Amplification Assay With Lateral Flow Dipstick Assay for Rapid Detection of 2019 Novel Coronavirus.

机构信息

School of Food Engineering and Biotechnology, Hanshan Normal University, Chaozhou, China.

Department of Medical Laboratory, Huizhou Central People's Hospital, Huizhou, China.

出版信息

Front Cell Infect Microbiol. 2021 Feb 1;11:613304. doi: 10.3389/fcimb.2021.613304. eCollection 2021.


DOI:10.3389/fcimb.2021.613304
PMID:33598439
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7882697/
Abstract

BACKGROUND: The emerging Coronavirus Disease-2019 (COVID-19) has challenged the public health globally. With the increasing requirement of detection for SARS-CoV-2 outside of the laboratory setting, a rapid and precise Point of Care Test (POCT) is urgently needed. METHODS: Targeting the nucleocapsid (N) gene of SARS-CoV-2, specific primers, and probes for reverse transcription recombinase-aided amplification coupled with lateral flow dipstick (RT-RAA/LFD) platform were designed. For specificity evaluation, it was tested with human coronaviruses, human influenza A virus, influenza B viruses, respiratory syncytial virus, and hepatitis B virus, respectively. For sensitivity assay, it was estimated by templates of recombinant plasmid and pseudovirus of SARS-CoV-2 RNA. For clinical assessment, 100 clinical samples (13 positive and 87 negatives for SARS-CoV-2) were tested quantitative reverse transcription PCR (RT-qPCR) and RT-RAA/LFD, respectively. RESULTS: The limit of detection was 1 copies/μl in RT-RAA/LFD assay, which could be conducted within 30 min at 39°C, without any cross-reaction with other human coronaviruses and clinical respiratory pathogens. Compared with RT-qPCR, the established POCT assay offered 100% specificity and 100% sensitivity in the detection of clinical samples. CONCLUSION: This work provides a convenient POCT tool for rapid screening, diagnosis, and monitoring of suspected patients in SARS-CoV-2 endemic areas.

摘要

背景:新型冠状病毒病 2019(COVID-19)的出现给全球公共卫生带来了挑战。随着实验室外对 SARS-CoV-2 检测需求的增加,迫切需要一种快速、准确的即时检测(POCT)方法。

方法:针对 SARS-CoV-2 的核衣壳(N)基因,设计了用于逆转录重组酶辅助扩增结合侧流层析试纸条(RT-RAA/LFD)平台的特异性引物和探针。为了评估特异性,分别用人冠状病毒、甲型流感病毒、乙型流感病毒、呼吸道合胞病毒和乙型肝炎病毒进行了测试。为了进行敏感性测定,使用 SARS-CoV-2 RNA 的重组质粒模板和假病毒进行了估计。为了进行临床评估,分别用定量逆转录 PCR(RT-qPCR)和 RT-RAA/LFD 对 100 份临床样本(SARS-CoV-2 阳性 13 份,阴性 87 份)进行了测试。

结果:在 RT-RAA/LFD 检测中,检测限为 1 拷贝/μl,可在 39°C 下 30 分钟内完成,与其他人类冠状病毒和临床呼吸道病原体无交叉反应。与 RT-qPCR 相比,该建立的 POCT 检测方法在检测临床样本时具有 100%的特异性和 100%的敏感性。

结论:本研究为 SARS-CoV-2 流行地区的疑似患者快速筛查、诊断和监测提供了一种便捷的 POCT 工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/b38306d8e84c/fcimb-11-613304-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/cc6f784a327b/fcimb-11-613304-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/9825a64d3079/fcimb-11-613304-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/b38306d8e84c/fcimb-11-613304-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/cc6f784a327b/fcimb-11-613304-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/9825a64d3079/fcimb-11-613304-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1f8/7882697/b38306d8e84c/fcimb-11-613304-g003.jpg

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本文引用的文献

[1]
Clinical Validation of Two Recombinase-Based Isothermal Amplification Assays (RPA/RAA) for the Rapid Detection of African Swine Fever Virus.

Front Microbiol. 2020-7-21

[2]
Establishment of reverse transcription recombinase-aided amplification-lateral-flow dipstick and real-time fluorescence-based reverse transcription recombinase-aided amplification methods for detection of the Newcastle disease virus in chickens.

Poult Sci. 2020-7

[3]
Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis.

Biosens Bioelectron. 2020-5-23

[4]
Single-copy sensitive, field-deployable, and simultaneous dual-gene detection of SARS-CoV-2 RNA via modified RT-RPA.

Cell Discov. 2020-5-28

[5]
[Source of the COVID-19 pandemic: ecology and genetics of coronaviruses (Betacoronavirus: Coronaviridae) SARS-CoV, SARS-CoV-2 (subgenus Sarbecovirus), and MERS-CoV (subgenus Merbecovirus).].

Vopr Virusol. 2020

[6]
Covid-19: Test and trace programmes are important but no silver bullet, say scientists.

BMJ. 2020-5-28

[7]
Reverse-Transcription Recombinase-Aided Amplification Assay for Rapid Detection of the 2019 Novel Coronavirus (SARS-CoV-2).

Anal Chem. 2020-7-10

[8]
Multiple-centre clinical evaluation of an ultrafast single-tube assay for SARS-CoV-2 RNA.

Clin Microbiol Infect. 2020-5-15

[9]
Rapid Detection of SARS-CoV-2 by Low Volume Real-Time Single Tube Reverse Transcription Recombinase Polymerase Amplification Using an Exo Probe with an Internally Linked Quencher (Exo-IQ).

Clin Chem. 2020-8-1

[10]
Rapid and sensitive detection of COVID-19 using CRISPR/Cas12a-based detection with naked eye readout, CRISPR/Cas12a-NER.

Sci Bull (Beijing). 2020-9-15

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