Eloiflin Roger, Pérez-Antón Elena, Camara Aïssata, Dujeancourt-Henry Annick, Boiro Salimatou, Djetchi Martial N, Traoré Mélika Barkissa, Koffi Mathurin, Kaba Dramane, Le Pennec Yann, Doukouré Bakary, Camara Abdoulaye Dansy, Kagbadouno Moïse, Campagne Pascal, Camara Mamadou, Jamonneau Vincent, Thévenon Sophie, Bart Jean-Mathieu, Glover Lucy, Rotureau Brice
INTERTRYP, Université de Montpellier, Cirad, IRD, Montpellier, France.
Trypanosome Molecular Biology Unit, Institut Pasteur, Université de Paris, Paris, France.
Elife. 2025 Sep 22;14:RP106823. doi: 10.7554/eLife.106823.
Animal African trypanosomosis (AAT), caused by protist parasites of the genus , puts upward of a million head of livestock at risk across 37 countries in Africa. The economic impact of AAT and the presence of human-infectious trypanosomes in animals place a clear importance on improving diagnostics for animal trypanosomes to map the distribution of the veterinary parasites and identify reservoirs of human-infectious trypanosomes. We have adapted the CRISPR-based detection toolkit SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) for trypanosomatid parasites responsible for AAT (SHERLOCK4AAT) including Pan-trypanosomatid, , , , , and assays. To test the applicability of this technique in the field, we analysed dried blood spots collected from 200 farm and 224 free-ranging pigs in endemic and historical human African trypanosomiasis foci in Guinea and Côte d'Ivoire, respectively. The results revealed that SHERLOCK4AAT can detect and discriminate between trypanosome species involved in multiple infections with a high sensitivity. 62.7% [58.1, 67.3] of pigs were found infected with at least one trypanosome species. , a human-infectious trypanosome, was found in one animal at both sites, highlighting the risk that these animals may act as persistent reservoirs. These data suggest that, due to their proximity to humans and their attractiveness to tsetse flies, pigs could act as sentinels to monitor circulation using the SHERLOCK4AAT toolbox.
动物非洲锥虫病（AAT）由布氏锥虫属的原生寄生虫引起，使非洲37个国家的100多万头牲畜面临风险。AAT的经济影响以及动物体内存在可感染人类的锥虫，凸显了改进动物锥虫诊断方法以绘制兽医寄生虫分布图并确定人类感染性锥虫宿主的重要性。我们已将基于CRISPR的检测工具包SHERLOCK（特异性高灵敏度酶促报告分子解锁）应用于导致AAT的锥虫寄生虫（SHERLOCK4AAT），包括泛锥虫、布氏冈比亚锥虫、布氏罗得西亚锥虫、伊氏锥虫、刚果锥虫、马媾疫锥虫和枯氏锥虫检测。为了测试该技术在实地的适用性，我们分别分析了从几内亚和科特迪瓦的地方性和历史性人类非洲锥虫病疫源地收集的200头农场猪和224头散养猪的干血斑。结果表明，SHERLOCK4AAT能够以高灵敏度检测和区分参与多重感染的锥虫种类。62.7%[58.1,67.3]的猪被发现感染了至少一种锥虫种类。在两个地点的一头动物中都发现了可感染人类的布氏罗得西亚锥虫，这凸显了这些动物可能作为持续宿主的风险。这些数据表明，由于猪与人类接近且对采采蝇有吸引力，它们可以作为哨兵，使用SHERLOCK4AAT工具箱监测锥虫传播。
