CRISPR/Cas12a and recombinase polymerase amplification-based rapid on-site nucleic acid detection of duck circovirus

文献类型： 外文期刊

第一作者： Liang, Qi-Zhang

作者： Liang, Qi-Zhang;Chen, Wei;Liu, Rong-Chang;Fu, Qiu-Ling;Fu, Guang-Hua;Cheng, Long-Fei;Chen, Hong-Mei;Jiang, Nan-Song;Huang, Yu;Chen, Wei;Zhu, Ting

作者机构：

关键词： Duck circovirus (DuCV); Recombinase polymerase amplification (RPA); Lateral flow strip (LFS); CRISPR/Cas12a; On-site detection

期刊名称：VIROLOGY JOURNAL （ 影响因子：3.8； 五年影响因子：3.7 ）

ISSN：

年卷期： 2024 年 21 卷 1 期

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收录情况： SCI

摘要： BackgroundDuck circovirus (DuCV) infections commonly induce immunosuppression and secondary infections in ducks, resulting in significant economic losses in the duck breeding industry. Currently, effective vaccines and treatments for DuCV have been lacking. Therefore, rapid, specific, and sensitive detection methods are crucial for preventing and controlling DuCV.MethodsA lateral flow strip (LFS) detection method was developed using recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 12a (Cas12a). The RPA-CRISPR/Cas12a-LFS targeted the DuCV replication protein (Rep) and was operated at 37 degrees C and allowed for visual interpretation without requiring sophisticated equipment.ResultsThe results revealed that the reaction time of RPA-CRISPR/Cas12a-LFS is only 45 min. This method achieved a low detection limit of 2.6 gene copies. Importantly, this method demonstrated high specificity and no cross-reactivity with six other avian viruses. In a study involving 97 waterfowl samples, the Rep RPA-CRISPR/Cas12a-LFS showed 100% consistency and agreement with real-time quantitative polymerase chain reaction.ConclusionThese findings underscored the potential of this user-friendly, rapid, sensitive, and accurate detection method for on-site DuCV detection.

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