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Rapid detection of Porcine circovirus 2 by recombinase polymerase amplification.

Authors
  • Wang, Jianchang1
  • Wang, Jinfeng1
  • Liu, Libing1
  • Li, Ruiwen1
  • Yuan, Wanzhe2
  • 1 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). , (China)
  • 2 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) [email protected] , (China)
Type
Published Article
Journal
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
Publication Date
September 2016
Volume
28
Issue
5
Pages
574–578
Identifiers
DOI: 10.1177/1040638716654201
PMID: 27493138
Source
Medline
Keywords
License
Unknown

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.

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