Establishment and application of a visual nucleic acid detection method for parvovirus

Parvoviruses pose a significant threat to the livestock breeding industry, leading to high morbidity and mortality rates. Rapid, accurate, and convenient detection methods are crucial for early intervention and control of parvovirus infection. This study established a novel visualized parvovirus detection method based on nucleic acid-mismatched enzyme detection—NMED. This method targets a conserved region of the Canine Parvo Virus (CPV) VP2 gene by utilizing specific fluorescent reporter probes and Recombinant Polymerase Amplification༈RPA༉ primers. Upon recognition of the target sequence by the reporter probe, T7 endonuclease Ⅰ (T7E Ⅰ) is activated, cleaving the probe and enabling detection under UV light or via the use of a fluorescence reader. Our research results show that we have successfully established a novel nucleic acid-based method for detecting CPV. This method can detect nucleic acid within 50 min. It exhibits high specificity, demonstrating no cross-reactivity with common canine pathogens. The method has a lower limit of detection of 1–10 copies/µL. The three replicates consistently yielded identical results, indicating high stability. Compared with qPCR, the NMED method achieved a concordance rate of 90.63%—93.75% for 32 suspected CPV samples. In conclusion, this study developed a convenient, efficient, visual nucleic acid detection method for CPV. The establishment of NMED has provided a new method for detecting parvovirus, which is highly important for the timely prevention and control of parvovirus infection and the reduction of livestock mortality.