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Research Article: Development of a rapid on-site nucleic acid detection method for new genotype muscovy duck parvovirus based on RPA-CRISPR/Cas12a

Date Published: 2025-08-26

Abstract:
New genotype Muscovy Duck Parvovirus (N-MDPV), a member of the Parvoviridae family, exhibits broad host tropism affecting Muscovy ducks, semi-Muscovy ducks, and white Kaiva duck. This pathogen causes severe morbidity and mortality in ducklings under 3?weeks of age, characterized by classic parvoviral lesions, beak atrophy, and growth retardation, posing substantial economic threats to China’s duck industry. To address diagnostic challenges, we developed an equipment-free detection platform targeting the conserved VP3 gene of N-MDPV. By integrating recombinase polymerase amplification (RPA) with CRISPR/Cas12a-mediated lateral flow strip (LFS) visualization, this method achieved isothermal amplification at 37°C within 35?min, eliminating dependency on thermocyclers. Validation experiments demonstrated exceptional sensitivity with a detection limit of 1.3 gene copies. Specificity testing revealed no cross-reactivity with eight common avian pathogens, confirming target exclusivity. Clinical validation using 98 field-collected duck tissue samples showed 98.98% concordance between our RPA-CRISPR/Cas12a-LFS and quantitative PCR. This study establishes the first CRISPR/Cas12a-based on-site diagnostic tool for N-MDPV, combining rapidity, sensitivity, accuracy and field-deployability.

Introduction:
Muscovy duck parvovirus (MDPV), the causative agent of Muscovy duck parvovirus disease (commonly termed “three-week disease”), is a highly contagious pathogen exclusively infecting Cairina moschata (Muscovy ducks) ( 1 , 2 ). It leads to open-mouthed breathing and diarrhea in infected ducks, and in fatal cases, pulmonary hemorrhage, pancreatic hemorrhage, and/or white necrotic spots, as well as duodenal mucosal bleeding. It primarily results in the illness and death of Muscovy ducks within 3?weeks of age ( 3 ). In…

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