Development of a real-time recombinase polymerase amplification assay for rapid and sensitive detection of porcine circovirus 2
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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 R2 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.
KeywordsClassical Swine Fever Virus Recombinase Polymerase Amplification Magnesium Acetate Lateral Flow Dipstick Recombinase Polymerase Amplification Assay
This work was supported by the Science and Technology Project Foundation of Hebei Province, P.R. China (Grant Number 16226604D). The funding agency had no role in the study design, the collection, analysis and interpretation of data, the writing of the report, or the decision to submit the article for publication.
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Conflict of interest
The authors declare that there is no conflict of interest.
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