In:
Journal of Veterinary Diagnostic Investigation, SAGE Publications, Vol. 28, No. 5 ( 2016-09), p. 574-578
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.
Type of Medium:
Online Resource
ISSN:
1040-6387
,
1943-4936
DOI:
10.1177/1040638716654201
Language:
English
Publisher:
SAGE Publications
Publication Date:
2016
detail.hit.zdb_id:
2265211-5
SSG:
22
Permalink