title: Rapid detection of Porcine circovirus 2 by recombinase polymerase amplification.
abstract: Porcine circovirus-associated disease, caused primarily by Porcine circovirus 2 (PCV-2), has become endemic in many pig-producing countries and has resulted in significant economic losses to the swine industry worldwide. Tests for PCV-2 infection include PCR, nested PCR, competitive PCR, and real-time PCR (rtPCR). Recombinase polymerase amplification (RPA) has emerged as an isothermal gene amplification technology for the molecular detection of infectious disease agents. RPA is performed at a constant temperature and therefore can be carried out in a water bath. In addition, RPA is completed in ~30 min, much faster than PCR, which usually takes >60 min. We developed a RPA-based method for the detection of PCV-2. The detection limit of RPA was 10(2) copies of PCV-2 genomic DNA. RPA showed the same sensitivity as rtPCR but was 10 times more sensitive than conventional PCR. Successful amplification of PCV-2 DNA, but not other viral templates, demonstrated high specificity of the RPA assay. This method was also validated using clinical samples. The results showed that the RPA assay had a diagnostic agreement rate of 93.7% with conventional PCR and 100% with rtPCR. These findings suggest that the RPA assay is a simple, rapid, and cost-effective method for PCV-2 detection, which could be potentially applied in clinical diagnosis and field surveillance of PCV-2 infection.
authors: Inspection and Quarantine Technical Center of Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang, Hebei, China (Jianchang Wang, Jinfeng Wang, Liu)College of Veterinary Medicine, Agricultural University of Hebei, Baoding, Hebei, China (Li, Yuan).
authors: Jianchang
authors: J
authors: Wang
authors: Inspection and Quarantine Technical Center of Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang, Hebei, China (Jianchang Wang, Jinfeng Wang, Liu)College of Veterinary Medicine, Agricultural University of Hebei, Baoding, Hebei, China (Li, Yuan).
authors: Jinfeng
authors: J
authors: Wang
authors: Inspection and Quarantine Technical Center of Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang, Hebei, China (Jianchang Wang, Jinfeng Wang, Liu)College of Veterinary Medicine, Agricultural University of Hebei, Baoding, Hebei, China (Li, Yuan).
authors: Libing
authors: L
authors: Liu
authors: Inspection and Quarantine Technical Center of Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang, Hebei, China (Jianchang Wang, Jinfeng Wang, Liu)College of Veterinary Medicine, Agricultural University of Hebei, Baoding, Hebei, China (Li, Yuan).
authors: Ruiwen
authors: R
authors: Li
authors: Inspection and Quarantine Technical Center of Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang, Hebei, China (Jianchang Wang, Jinfeng Wang, Liu)College of Veterinary Medicine, Agricultural University of Hebei, Baoding, Hebei, China (Li, Yuan) yuanwanzhe2015@126.com.
authors: Wanzhe
authors: W
authors: Yuan
mesh_terms: D000818:Animals; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D017925:Circovirus / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D045522:Recombinases; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology
chemical_list: D045522:Recombinases
keywords: Porcine circovirus-2; recombinase polymerase amplification