CRISPR-Cas13a-based detection method for avian influenza virus

文献类型： 外文期刊

第一作者： Wu, Yuhan

作者： Wu, Yuhan;Zhan, Jiaxing;Shan, Zhaomeng;Liu, Yining;Li, Yan;Wang, Yixin;Liu, Zhe;Wen, Xuexia;Li, Yanbing;Wang, Xiurong

作者机构：

关键词： avian influenza virus; recombinase polymerase amplification; CRISPR-cas13a; lateral flow dipstick assay; universal detection

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.2； 五年影响因子：6.2 ）

ISSN：

年卷期： 2023 年 14 卷

页码：

收录情况： SCI

摘要： Avian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a based platform was developed to detect AIV. This novel method was developed to specifically detect H1-H16 subtypes of AIV with fluorescence and lateral flow-based readouts and exhibited no cross-reactivity with Newcastle disease virus, avian infectious bronchitis virus, or infectious bursal disease virus. The limit of detection was determined to be 69 and 690 copies/mu L using fluorescence and lateral flow as readouts, respectively. The developed assay exhibited 100% consistency with quantitative real-time polymerase chain reaction in detecting clinical samples. The heating of unextracted diagnostic samples to obliterate nuclease treatment was introduced to detect viral RNA without nucleic acid extraction. Single-step optimization was used to perform reverse transcription, recombinase polymerase amplification, and CRISPR-Cas13a detection in a tube. These advances resulted in an optimized assay that could specifically detect AIV with simplified procedures and reduced contamination risk, highlighting the potential to be used in point-of-care testing.

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