Innovative Technology

Innovation

Focusing on the research and development and application of isothermal nucleic acid amplification technology

Development of a Real-Time Fluorescent RAA Assay for Porcine Circovirus Type 4 – Chen Weiyue

  • File size: 18.3MB

To rapidly detect Porcine Circovirus Type 4 (PCV4), this study designed primers and probes based on the conserved region of the Cap gene fragment of PCV4, and developed a recombinase-mediated isothermal amplification fluorescence assay (RAA) for PCV4. The developed RAA method was then applied to test 40 porcine tissue samples. The results showed that this method can specifically detect PCV4 within 20 minutes under constant temperature conditions at 42°C. When using nucleic acids from viruses such as Porcine Circovirus Type 2 (PCV2), Porcine Circovirus Type 3 (PCV3), Porcine Epidemic Diarrhea Virus (PEDV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and Porcine Transmissible Gastroenteritis Virus (TGEV) as templates, the amplification reactions yielded negative results. The method demonstrated a low detection limit of 7.42 × 10¹ copies/μL, indicating high sensitivity. When the established RAA method and conventional PCR were used to analyze the same porcine tissue samples, neither method detected any viral nucleic acids, suggesting that the current prevalence of PCV4 in pig farms is relatively low. In summary, the real-time fluorescent RAA method for PCV4 established in this study is rapid, simple, highly specific, and sensitive, making it suitable for clinical screening and epidemiological studies of PCV4.

Previous page

None

Establishment of a Fluorescence Detection Method for RAA, a Recombinase-Aided Isothermal Amplification Technique for Porcine Eperythrozoon—China Animal Health and Veterinary Medicine

Next page

Previous page

None

Next page

Establishment of a Fluorescence Detection Method for RAA, a Recombinase-Aided Isothermal Amplification Technique for Porcine Eperythrozoon—China Animal Health and Veterinary Medicine