Despite its extensive presence among grazing ruminants, dicrocoeliosis, also known as ‘small liver fluke’ disease, is poorly known and often underestimated by researchers and practitioners in many countries. The accurate identification and prepatent diagnosis of Dicrocoelium dendriticum infection is an essential prerequisite for its prevention and control. In the present study, the morphologically identified specimens isolated from the bile ducts of sheep (Ovis aries) were validated through molecular data. The sequence analysis of the second internal transcribed spacer (ITS-2) of our isolates showed a high degree of similarity with D. dendriticum using the BLAST function of the National Center for Biotechnology Information (NCBI). The phylogenetic analysis of our isolates showed a close relationship with previously described D. dendriticum isolates from different countries. The antigenic profiles of somatic and excretory/secretory (E/S) antigens of D. dendriticum were revealed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting using sera from sheep naturally infected with D. dendriticum. By SDS–PAGE, 16 distinct bands were revealed from crude somatic fraction. Immunoblotting analysis of these proteins with positive sera exhibited six seroreactive bands ranging from 27 to 130 kDa. Among these, the 84 and 130 kDa bands were quite specific, with high diagnostic specificity and sensitivity. The E/S fraction comprised nine distinct bands, as revealed by SDS–PAGE analysis. Immunoblotting analysis of these proteins with positive sera exhibited five antigenic bands ranging from 27 to 130 kDa. Among these, the 130 kDa band was found to be quite specific, with high diagnostic specificity and sensitivity. The present study concludes that the protein bands of 84 and 130 kDa in somatic fraction and 130 kDa in E/S fraction can be used for the immunodiagnostic purpose for this economically important parasite, which may also encourage further studies regarding their vaccine potential.

中文翻译：

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从喜马拉雅西北地区绵羊分离的树突状双球菌的分子特征和免疫诊断

尽管它在放牧反刍动物中广泛存在，但也被称为“小肝吸虫病”的双球囊虫病鲜为人知，而且许多国家的研究人员和从业人员常常低估了它。准确识别和预诊断树突状重腔菌感染是其预防和控制的必要前提。在本研究中，从绵羊胆管中分离出的经形态鉴定的标本（绵羊) 通过分子数据进行了验证。我们分离株的第二个内部转录间隔区（ITS-2）的序列分析显示与D.树突状使用国家生物技术信息中心 (NCBI) 的 BLAST 功能。我们的分离株的系统发育分析显示与先前描述的密切相关D.树突状来自不同国家的隔离。体细胞和排泄/分泌 (E/S) 抗原的抗原谱D.树突状通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳 (SDS-PAGE) 和使用自然感染羊血清的免疫印迹法显示D.树突状。通过 SDS-PAGE，从粗体细胞组分中显示出 16 个不同的条带。这些具有阳性血清的蛋白质的免疫印迹分析显示出六个血清反应带，范围从 27 到 130 kDa。其中，84 和 130 kDa 条带特异性较高，诊断特异性和敏感性较高。如 SDS-PAGE 分析所示，E/S 部分包含九个不同的条带。这些具有阳性血清的蛋白质的免疫印迹分析显示出 5 个 27 至 130 kDa 的抗原带。其中，发现 130 kDa 条带具有很高的特异性，具有很高的诊断特异性和敏感性。本研究得出结论，体细胞部分中 84 和 130 kDa 的蛋白质条带和 E/S 部分中 130 kDa 的蛋白质条带可用于这种经济上重要的寄生虫的免疫诊断目的，