Knapp Jenny, Da Silva Abdou Malik, Courquet Sandra, Millon Laurence
National Reference Centre for Echinococcoses, Department of Parasitology-Mycology, University Hospital of Besançon, 25030 Besançon, France.
Laboratoire Chrono-Environnement, UMR CNRS 6249 Université Bourgogne-Franche-Comté, 16 Route de Gray, 25030 Besançon, France.
Pathogens. 2021 Oct 8;10(10):1296. doi: 10.3390/pathogens10101296.
The genetic diversity of the parasite , the infectious agent of alveolar echinococcosis, is generally assessed on adult worms after fox necropsy. We aimed to investigate polymorphism through the microsatellite EmsB marker using a noninvasive approach. We tested batches of isolated eggs (1, 5, and 10) from 19 carnivore fecal samples collected in a rural town located in a highly endemic area in France to determine the best strategy to adopt using a minimal quantity of parasite DNA while avoiding genetic profile overlapping in the analysis. Several molecular controls were performed to formally identify the Taeniidae eggs. In total, 112 egg batches were isolated and 102 EmsB electrophoregrams were obtained in duplicate. Quality sorting was performed through the Pearson correlation coefficient (r) between each EmsB duplicate. Forty-nine batches with r > 0.9 remained in the analysis, mainly 5- or 10-egg batches. Three EmsB profiles were emphasized by hierarchical clustering and matched those from human lesions and adult worms previously genotyped and collected in the same area. We show that the genetic diversity of the parasite can be assessed from isolated eggs in a spatiotemporal context using a noninvasive approach.
泡型包虫病的病原体——寄生虫的遗传多样性通常是在狐狸尸检后对成虫进行评估。我们旨在采用非侵入性方法，通过微卫星EmsB标记研究多态性。我们对从法国一个高度流行地区的乡村小镇采集的19份食肉动物粪便样本中的分离虫卵批次（1个、5个和10个）进行了检测，以确定在分析中使用最少数量的寄生虫DNA同时避免基因图谱重叠的最佳策略。进行了多项分子对照以正式鉴定带绦虫卵。总共分离出112个虫卵批次，并获得了102份EmsB电泳图的复制品。通过每个EmsB复制品之间的皮尔逊相关系数（r）进行质量分选。49个r>0.9的批次进入分析，主要是5个或10个虫卵的批次。通过层次聚类突出了三种EmsB图谱，它们与先前在同一地区进行基因分型并收集的人类病变和成虫的图谱相匹配。我们表明，使用非侵入性方法可以在时空背景下从分离的虫卵中评估寄生虫的遗传多样性。
