重组酶聚合酶扩增
生物
病毒学
猪圆环病毒
清脆的
核酸
PCR的应用
圆环病毒
聚合酶链反应
多重位移放大
聚合酶
分子生物学
数字聚合酶链反应
DNA
遗传学
病毒
基因
DNA提取
作者
Qi-Zhang Liang,Wei Chen,Rong-Chang Liu,Qiuling Fu,Guanghua Fu,Longfei Cheng,Hongmei Chen,Nansong Jiang,Ting Zhu,Yu Huang
标识
DOI:10.1186/s12985-024-02577-7
摘要
Duck 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. A 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 ℃ and allowed for visual interpretation without requiring sophisticated equipment. The 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. These findings underscored the potential of this user-friendly, rapid, sensitive, and accurate detection method for on-site DuCV detection.
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