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H9N2 avian influenza virus diagnostics utilizing specific high-sensitivity enzymatic molecular system termed RPA-based CRISPR-Cas13a

文献类型： 外文期刊

作者： He, Dalin 1 ; Zhao, Saisai 1 ; Wang, Fangfang 1 ; Wu, Bingrong 1 ; Wei, Feng 1 ; Zhao, Yubo 1 ; Wei, Xinhui 1 ; Ren, Hui 1 ; Zhang, Meijuan 1 ; Fan, Yaru 1 ; Zhang, Jiahao 1 ; Yu, Shumin 1 ; Tang, Yi 2 ; Diao, Youxiang 1 ;

作者机构： 1.Shandong Agr Univ, Coll Vet Med, Tai An 271018, Shandong, Peoples R China

2.Chinese Acad Agr Sci, Inst Anim Sci, Beijing 10091, Peoples R China

关键词： Recombinase polymerase amplification; CRISRPR/Cas13; Lateral flow assay

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）

ISSN： 0141-8130

年卷期： 2025 年 301 卷

页码：

收录情况： SCI

摘要： H9N2 avian influenza virus (AIV), a major pathogen causing respiratory infections in poultry, poses a significant threat to the poultry industry and human health. Early detection and control of H9N2 infections are essential for minimizing economic losses and preventing potential zoonotic transmission. A novel CRISPR-Cas family member called CRISPR-Cas13a comprises the CRISPR RNA (crRNA) and Cas13a nuclease. Through the crRNA-based reprogramming of Cas13a, a platform for sensing RNAs specifically is available. In this study, we developed a RPA-based CRISPR-Cas13a diagnostic method for rapid detection of the H9N2 AIV. The results demonstrated that at a limit of 10 copies/mu L and 102 copies/mu L could be detected within 50 min, by fluorescence detection and lateral flow strip, respectively, offering a highly sensitive method for H9N2 detection. This method exhibited excellent specificity, distinguishing H9N2 from other pathogens. Furthermore, the RPA-Cas13a-based detection system was tested on clinical samples, showing comparable performance to RT-qPCR. The detection results were visualized using either lateral flow assays or fluorescence, making it a suitable tool for on-site, field-deployable diagnostics. In a word, this RPA-Cas13a diagnostic approach offers high reliability, sensitivity, and specificity, with promising potential for rapidly detecting H9N2 and other viral pathogens in clinical and food safety applications.

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