数字农科院2.0

Rapid visual detection of Moniezia spp. in sheep feces via Recombinase Polymerase Amplification-Lateral Flow Dipstick (RPA-LFD) assay

文献类型： 外文期刊

作者： Shaohua Zhang;Yeping Zhao;Weijia Liang;Shuai Wang;Xiu Cui;Haohan Zhu;Yueyue Zhang;Xiaolei Liu;Huimin Li;Wenjie Mu;Aijiang Guo

作者机构：

关键词： Lateral flow dipstick (LFD);Monieziasis;Recombinase polymerase amplification (RPA);Visual detection;β-tubulin gene

期刊名称： Veterinary Parasitology

ISSN： 0304-4017

年卷期： 2025 年 339 卷

页码：

收录情况： SCIE（2025版）

摘要： Monieziasis is a prevalent issue in small ruminant husbandry, primarily caused by Moniezia expansa and M. benedeni in China. There is a critical need for highly sensitive methods for disease surveillance and prevention. In this study, we developed a visual assay combining recombinase polymerase amplification (RPA) with a lateral flow strip (RPA-LFD) for the rapid detection of Moniezia spp. in sheep fecal samples. Seven primer/probe sets were designed based on a specific fragment of the M. expansa β-tubulin gene (MTUB). The RPA-LFD assay performed optimally under reaction conditions of 39 ℃ for 15 min, with a primer-to-probe ratio of 4:0.6. The optimized method demonstrated high specificity for Moniezia spp., with no cross-reactivity with genomic DNA from other common gastrointestinal parasites in ruminant livestock. The detection limit was 10 copies/μL of plasmid DNA or 10 pg/μL of M. expansa genomic DNA per reaction. When compared to PCR using clinical and sheep-simulated samples, the RPA-LFD assay exhibited equivalent detection capability, achieving 95.7 % consistency (K value = 0.939, p < 0.001). These results suggest that the RPA-LFD method is a reliable and portable diagnostic tool for routine screening, monitoring, and rapid confirmation of monieziasis in sheep flocks, particularly in endemic areas and remote regions.

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