The species identification of tick vectors of Crimean-Congo hemorrhagic fever virus (CCHFV), especially Hyalomma (H.) species, is a prerequisite to understand the eco-epidemiology of this disease and to reveal vector and virus reservoir species. However, the morphologic species discrimination can be difficult for damaged or blood-fed ticks and in case of species intercrosses. Therefore, we used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and restriction fragment length polymorphism (RFLP) analysis to distinguish the most common Hyalomma species from sub-Saharan Africa (H. truncatum, H. rufipes and H. dromedarii). Within the last years, MALDI-TOF MS analysis based on tick leg proteins has been shown to be a reliable method to distinguish several tick species. For this purpose, a reference spectral library of several European, American and African tick species was established. In this study, six different Hyalomma species were tested, all of which were all clearly distinguishable by mass spectrometric analyses. Moreover, MALDI TOF- MS was able to confirm morphologic findings where sequencing provided ambiguous results. In addition, a polymerase chain reaction (PCR) based on the CO1 gene amplification of ticks has been developed for the unequivocal species identification by amplicon sequencing and specific restriction endonuclease cleavage pattern analysis. RFLP proved to be a feasible auxiliary discrimination tool for selected Hyalomma species when access to sequencing methods is not available, as for instance during field studies.

克里米亚-刚果出血热病毒（CCHFV），尤其是透明质酸（H.）物种的壁虱载体的物种识别是了解该病的生态流行病学和揭示载体和病毒库物种的先决条件。但是，形态特征的物种识别对于受损的或血腥的壁虱以及物种交叉时可能很困难。因此，我们使用基质辅助激光解吸/电离时间飞行质谱（MALDI-TOF MS）和限制性片段长度多态性（RFLP）分析，以区分最常见璃眼从撒哈拉以南非洲的物种（H.元宝，rufipes和H. dromedarii）。在过去的几年中，基于tick腿蛋白的MALDI-TOF MS分析已被证明是区分几种tick种的可靠方法。为此，建立了几种欧洲，美洲和非洲壁虱物种的参考光谱库。在这项研究中，六种不同的玻尿酸对这些物种进行了测试，所有这些都可以通过质谱分析清楚地区分。此外，MALDI TOF-MS能够确认测序结果不明确的形态学发现。另外，已经开发了基于tick的CO1基因扩增的聚合酶链反应（PCR），用于通过扩增子测序和特异性限制性核酸内切酶切割模式分析来明确鉴定物种。当无法获得测序方法时，例如在田野研究期间，RFLP被证明是针对特定的透明膜物种的可行辅助鉴别工具。