Recombinant Polymerase Amplification Coupled with CRISPR/Cas12a Detection System for Rapid Visual Detection of Porcine Circovirus 3

Autor: Genghong Jiang, Xiaoyu Yang, Zhaoyang Li, Jingyu Mao, Penghui Zeng, Dedong Wang, Zhi Wu, Changzhe Liu, Yonghui Qiu, Yongqiu Cui, Jianwei Zhou, Jue Liu, Lei Hou
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Animals, Vol 14, Iss 17, p 2527 (2024)
Druh dokumentu: article
ISSN: 2076-2615
DOI: 10.3390/ani14172527
Popis: The porcine circovirus type 3 (PCV3) infection is an emerging disease associated with clinical signs of porcine dermatitis and nephropathy syndrome (PDNS)-like clinical signs. Currently, there is a lack of effective vaccines and therapeutics against this disease. Therefore, rapid, effective, sensitive, and specific detection methods are crucial for the timely identification, prevention, and control of PCV3. In this study, we developed one- and two-pot visual detection methods for PCV3 using a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a detection system combined with recombinase polymerase amplification (RPA). These two methods demonstrated no cross-reactivity with eight other swine viruses and exhibited minimum detection limits of five and two copies of viral DNA, respectively, revealing their high specificity and sensitivity. During a clinical sample detection within 30 min, the coincidence rates between the one- and two-pot detection methods and real-time quantitative polymerase chain reaction (qPCR) were 100%. In conclusion, both one- and two-pot RPA-CRISPR/Cas12a detection methods have significant potential for the rapid, sensitive, and specific visual detection of PCV3.
Databáze: Directory of Open Access Journals
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje