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

文献类型： 外文期刊

第一作者： Liang, Qizhang

作者： Liang, Qizhang;Chen, Wei;Wang, Weiwei;Liu, Rongchang;Fu, Qiuling;Fu, Guanghua;Cheng, Longfei;Jiang, Nansong;Chen, Hongmei;Huang, Yu

作者机构：

关键词： N-MDPV; RPA; LFS; CRISPR/Cas12a; on-site detection

期刊名称：FRONTIERS IN VETERINARY SCIENCE （ 影响因子：2.9； 五年影响因子：3.3 ）

ISSN：

年卷期： 2025 年 12 卷

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收录情况： SCI

摘要： 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 degrees 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.

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