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- Title
Development of a real-time recombinase polymerase amplification assay for rapid and sensitive detection of porcine circovirus 2.
- Authors
Wang, Jianchang; Wang, Jinfeng; Liu, Libing; Yuan, Wanzhe
- Abstract
Porcine diseases associated with porcine circovirus 2 (PCV-2) infection have resulted in significant economic losses worldwide. A real-time recombinase polymerase amplification (RPA) assay was developed to detect PCV-2 using primers and an exo probe specific for the ORF2 gene. The reaction process can be completed in 20 min at 38 °C. The assay only detects PCV-2, as there was no cross-reaction with other pathogens important in pigs. Using the PCV-2 genomic DNA as template, the analytical sensitivity of the real-time RPA was 103 copies. The assay performance was evaluated by testing 38 field samples and compared with real-time PCR. The two assays demonstrated a 100% diagnostic agreement, and PCV-2 DNA was detected in 26 samples. The R value of real-time RPA and real-time PCR was 0.954 by linear regression analysis. The real-time RPA assay provides an alternative tool for rapid, simple, and reliable detection of PCV-2, especially in remote and rural areas.
- Subjects
RECOMBINASES; CIRCOVIRUS diseases; DNA primers; GENE amplification; REGRESSION analysis; POLYMERASE chain reaction
- Publication
Archives of Virology, 2017, Vol 162, Issue 8, p2293
- ISSN
0304-8608
- Publication type
Article
- DOI
10.1007/s00705-017-3368-3