Waterfowl parvoviruses, including goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV), can cause seriously diseases in geese and ducks. Developing a fast and precise diagnosis assay for these two parvoviruses is particularly important. A duplex SYBR Green I-based quantitative real-time PCR assay was developed for the simultaneous detection and differentiation of GPV and MDPV. The assay yielded melting curves with specific single peak (Tm = 87.3 ± 0.26 °C or Tm = 85.4 ± 0.23 °C) when GPV or MDPV was evaluated, respectively. When both parvoviruses were assessed in one reaction, melting curves with specific double peaks were yielded. This duplex quantitative RT-PCR can be used to rapid identify of GPV and MDPV in field cases and artificial trials, which make it a powerful tool for diagnosing, preventing and controlling waterfowl parvovirus infections.

Short beak and dwarfism syndrome virus (SBDSV) is the causative agent of short beak and dwarfism syndrome (SBDS), which is characterized by beak atrophy and dwarfism. SBDS has caused substantial economic losses for waterfowl husbandry industries. To address the urgent need for rapid and accurate SBDSV diagnostics, the study developed a dual-mode detection platform integrating recombinase-aided amplification (RPA) with CRISPR/Cas12a-mediated fluorescence and lateral flow strip readouts. The optimized assay achieved a detection limit of 10 copies/μL. Notably, the platform demonstrated superior specificity to distinguish SBDSV from genetically related Muscovy duck-origin goose parvovirus (GPV) and classical GPV, a distinction unachievable by qPCR. Clinical validation using 36 field samples confirmed 100% concordance with qPCR and indirect immunofluorescence assays, with no cross-reactivity against other common duck pathogens. This innovative detection system provides a robust toolkit for field-deployable SBDSV surveillance and lays a solid foundation for the development of novel diagnostic methodologies applicable to various waterfowl-related pathogens.

Goose parvovirus (GPV) remains a major pathogen in waterfowl production, threatening animal health and industry sustainability. In this study, a real-time recombinase polymerase amplification (RPA) assay was developed for rapid and specific detection of GPV. The assay employed a primer‒probe system targeting the VP1 gene and operated under isothermal conditions at 40°C. Analytical sensitivity was determined using serial dilutions of plasmid DNA, establishing a detection limit of 3.0 × 10¹ copies/μL with high linearity (R² = 0.99). No cross-reactivity was observed with other common avian pathogens, confirming the specificity of the assay. Clinical validation using 53 field samples demonstrated 98.1 % overall agreement with real-time qPCR. This method requires minimal equipment and yields results within 20 min, making it more suitable for field applications than conventional PCR- or qPCR-based platforms. These findings indicate that the developed real-time RPA assay is a rapid, sensitive, and reliable diagnostic tool for GPV detection, with potential utility in epidemiological surveillance and disease control in waterfowl populations.