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： Zhang, Shaohua 1 ; Zhao, Yeping 1 ; Liang, Weijia 1 ; Wang, Shuai 1 ; Cui, Xiu 1 ; Zhu, Haohan 1 ; Zhang, Yueyue 1 ; Liu, Xiaolei 2 ; Li, Huimin 1 ; Mu, Wenjie 1 ; Guo, Aijiang 1 ;
作者机构： 1.Chinese Acad Agr Sci, Lanzhou Vet Res Inst, State Key Lab Anim Dis Control & Prevent, Key Lab Vet Parasitol Gansu Prov, Lanzhou, Peoples R China
2.Jilin Univ, Coll Vet Med, State Key Lab Diag & Treatment Severe Zoonot Infec, Minist Educ,Key Lab Zoonosis Res, Changchun, Peoples R China
3.Gansu Agr Univ, Coll Vet Med, Lanzhou, Peoples R China
关键词： Monieziasis; Recombinase polymerase amplification (RPA); Lateral flow dipstick (LFD); Visual detection; beta-tubulin gene
期刊名称：VETERINARY PARASITOLOGY （ 影响因子：2.2； 五年影响因子：2.4 ）
ISSN： 0304-4017
年卷期： 2025 年 339 卷
页码：
收录情况： SCI
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 beta-tubulin gene (MTUB). The RPA-LFD assay performed optimally under reaction conditions of 39 degrees C 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/mu L of plasmid DNA or 10 pg/mu L 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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