Zhao Xiaoyan, Zheng Ximing, Yang Xiyong, Guo Qi, Zhang Yi, Lou Jun
Department of Clinical Laboratory, Zhumadian Central Hospital, Zhumadian City, Henan Province, China.
Laboratory of Virology, Beijing Key Laboratory of Etiology of Viral Diseases in Children, Capital Institute of Pediatrics, Beijing, China.
China CDC Wkly. 2024 Sep 13;6(37):946-952. doi: 10.46234/ccdcw2024.198.
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.
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.
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.
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.
本研究旨在利用基质蛋白基因的保守区域开发一种快速、可视化的PCR-CRISPR/Cas12-LFD方法来检测甲型流感病毒。
我们设计了通用降解引物和靶向流感病毒（IFV）保守基质蛋白基因的成簇规律间隔短回文重复序列RNA（CRISPR RNA，crRNA），并与侧向流动试纸条（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确定其敏感性和特异性。
所开发的甲型流感病毒（IAV）核酸检测方法灵敏度为10拷贝/μL，与其他呼吸道病原体无交叉反应。对82份临床样本的评估显示，与荧光聚合酶链反应（PCR）结果高度一致，kappa值为0.95。
已成功建立了一种高灵敏度和特异性的PCR-CRISPR/Cas12-LFD方法用于检测甲型流感病毒,为其诊断提供了有力工具，并有助于该病毒的预防和控制。
