Department of Botany and Zoology, Faculty of Science, Masaryk University, Brno, 611 37, Czech Republic.
Department of Zoology and Fisheries, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague, 165 00, Czech Republic.
Sci Rep. 2020 Sep 29;10(1):15984. doi: 10.1038/s41598-020-73037-9.
Among gastrointestinal nematodes, haematophagous strongylids Haemonchus contortus and Ashworthius sidemi belong to the most pathogenic parasites of both domestic and wild ruminants. Correct identification of parasitic taxa is of crucial importance in many areas of parasite research, including monitoring of occurrence, epidemiological studies, or testing of effectiveness of therapy. In this study, we identified H. contortus and A. sidemi in a broad range of ruminant hosts that occur in the Czech Republic using morphological/morphometric and molecular approaches. As an advanced molecular method, we employed qPCR followed by High Resolution Melting analysis, specifically targeting the internal transcribed spacer 1 (ITS-1) sequence to distinguish the two nematode species. We demonstrate that High Resolution Melting curves allow for taxonomic affiliation, making it a convenient, rapid, and reliable identification tool.
在胃肠道线虫中，寄生性的强旋毛线虫（Haemonchus contortus）和半细线虫（Ashworthius sidemi）属于对家养和野生反刍动物最具致病性的寄生虫之一。在寄生虫研究的许多领域，包括寄生虫发生的监测、流行病学研究或治疗效果的测试，正确识别寄生分类群至关重要。在这项研究中，我们使用形态学/形态计量学和分子方法，从分布于捷克共和国的广泛反刍动物宿主中鉴定出了 H. contortus 和 A. sidemi。作为一种先进的分子方法，我们采用了 qPCR 结合高分辨率熔解曲线分析（HRM），该方法专门针对内部转录间隔区 1（ITS-1）序列，以区分这两种线虫。我们证明了 HRM 曲线可以进行分类归属，使其成为一种方便、快速和可靠的鉴定工具。
