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

猪圆环病毒4型实时荧光RAA检测方法的建立_陈伟月

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To rapidly detect Porcine Circovirus Type 4 (PCV4), this study designed primers and probes targeting the conserved region of the PCV4 Cap gene, successfully establishing a Recombinase-Aided Amplification (RAA) assay with fluorescence detection. The method was then applied to test 40 porcine tissue samples. Results showed that the assay can specifically detect PCV4 within 20 minutes under constant temperature conditions at 42°C, with no amplification detected 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. Additionally, the assay demonstrated an impressive detection limit as low as 7.42 × 10¹ copies/μL, indicating high sensitivity. When both the established RAA method and conventional PCR were used to analyze the same set of porcine tissue samples, neither technique yielded positive results, suggesting that the current prevalence of PCV4 in pig farms remains relatively low. In conclusion, the PCV4 real-time fluorescent RAA assay developed in this study is rapid, simple, highly specific, and remarkably sensitive, making it an ideal tool for clinical screening and epidemiological studies of PCV4.