Development of a real-time recombinase polymerase amplification assay for rapid and sensitive detection of porcine circovirus 2

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 c...

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Bibliographic Details
Published in:Archives of virology 2017-08, Vol.162 (8), p.2293-2296
Main Authors: Wang, Jianchang, Wang, Jinfeng, Liu, Libing, Yuan, Wanzhe
Format: Article
Language:English
Subjects:
Animals
Biomedical and Life Sciences
Biomedicine
Circoviridae Infections - diagnosis
Circoviridae Infections - veterinary
Circovirus - isolation & purification
Cross-reaction
DNA, Viral - isolation & purification
Infectious Diseases
Linear Models
Medical Microbiology
Molecular Diagnostic Techniques - methods
Molecular Diagnostic Techniques - veterinary
Nucleic Acid Amplification Techniques - methods
Nucleic Acid Amplification Techniques - veterinary
Orf2 gene
Original Article
Polymerase chain reaction
Primers
Real-Time Polymerase Chain Reaction - methods
Real-Time Polymerase Chain Reaction - veterinary
Recombinase
Sensitivity and Specificity
Swine - virology
Swine Diseases - diagnosis
Virology
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Summary: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 2 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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-017-3368-3