Establishment of a Lateral Flow Dipstick Detection Method for Influenza A Virus Based on CRISPR/Cas12a System.

IF 2.9 Q1 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH 中国疾病预防控制中心周报 Pub Date : 2024-09-13 DOI:10.46234/ccdcw2024.198

Xiaoyan Zhao, Ximing Zheng, Xiyong Yang, Qi Guo, Yi Zhang, Jun Lou

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Abstract

Objective: This study aimed to develop a rapid, visual PCR-CRISPR/Cas12-LFD method for detecting influenza A by utilizing the conserved region of the matrix protein gene.

Method: We crafted universal degradation primers and clustered regularly interspaced short palindromic repeats RNA (CRISPR RNA, crRNA) targeting the conserved matrix protein gene of the influenza virus (IFV), integrated with lateral flow dipstick (LFD) technology. This new PCR-CRISPR/Cas12-LFD approach was designed to determine its sensitivity and specificity through the analysis of various clinical samples collected in 2023.

Results: The developed nucleic acid assay for influenza A viruses (IAV) demonstrated a sensitivity of 10 copies/μL without cross-reactivity with other respiratory pathogens. Evaluation of 82 clinical samples showed high concordance with results from fluorescent Polymerase Chain Reaction (PCR), achieving a kappa value of 0.95.

Conclusion: A highly sensitive and specific PCR-CRISPR/Cas12-LFD method has been successfully established for the detection of influenza A, offering a robust tool for its diagnosis and aiding in the prevention and control of this virus.

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基于CRISPR/Cas12a系统的甲型流感病毒侧流浸量杆检测方法的建立

研究目的本研究旨在利用基质蛋白基因的保守区，开发一种快速、可视的PCR-CRISPR/Cas12-LFD方法来检测甲型流感：方法：我们针对流感病毒（IFV）的保守基质蛋白基因制作了通用降解引物和簇状规律性间隔短回文重复序列RNA（CRISPR RNA，crRNA），并将其与侧向流动点滴（LFD）技术相结合。这种新的 PCR-CRISPR/Cas12-LFD 方法旨在通过分析 2023 年收集的各种临床样本来确定其灵敏度和特异性：结果：所开发的甲型流感病毒（IAV）核酸检测方法的灵敏度为 10 个拷贝/μL，与其他呼吸道病原体无交叉反应。对 82 份临床样本的评估结果显示，与荧光聚合酶链反应（PCR）的结果高度一致，卡帕值达到 0.95：高灵敏度和特异性的 PCR-CRISPR/Cas12-LFD 方法已成功用于检测甲型流感，为甲型流感的诊断提供了强有力的工具，有助于该病毒的预防和控制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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中国疾病预防控制中心周报

CiteScore

5.90

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