Novel Muscovy DuckParvo virus Fluorescent RPA Constant Temperature Rapid Establishment and application of detection methods

【Objective】 Provide a visual and rapid detection technology for the rapid detection of the New-genotype Muscovy duck Parvovirus at the grassroots level.【Methods】 This study targets the conserved fragment of the VP3 gene of the New-genotype Muscovy duck Parvovirus. Recombinant enzyme polymerase amplification (RPA) technology was used and specific RPA amplification primers were designed. Combined with EXO fluorescent probes, an accurate and efficient isothermal rapid detection method for the New-genotype Muscovy duck Parvovirus RPA was established. The sensitivity and specificity of the detection method were evaluated, and compared with traditional PCR methods and virus isolation and identification methods.【Results】 This detection method can detect 10 fg/ μL The viral nucleic acid of L has high sensitivity; Only specific amplification of the New-genotype Muscovy duck Parvovirus was performed, and there was no cross reaction with duck adenovirus type 3, duck adenovirus type 4, duck circovirus, duck plague virus, duck viral hepatitis virus, duck tambusu virus, and new duck reovirus, indicating good specificity; Using the RPA method established in this study, the PCR method established earlier, and the virus isolation and identification method, 38 clinical collected duck tissue samples were tested, and the results showed positive rates of 36.8% (14/38), 36.8% (14/38), and 31.6% (12/38), respectively; And the positive samples detected by RPA were tested positive by PCR and virus isolation and identification methods, with a compliance rate of 100%. The compliance rate with PCR in clinical sample testing is 100%. 【Conclusion】 This method can be well applied to grassroots areas lacking corresponding detection equipment for large-scale clinical sample detection of the New-genotype Muscovy duck Parvovirus, providing technical means for visual rapid detection and epidemiological investigation of the novel muscovy duck parvovirus.